Clinical Focus

  • Anesthesia

Academic Appointments

Administrative Appointments

  • Asst. Clinical Director, Byers Eye Institute, Anesthesiology (2013 - Present)

Honors & Awards

  • Best Scientific Presentation, Western Anesthesia Residents Conference (5/2012)
  • Best Clinical Abstract, Western Anesthesia Residents Conference (5/2012)
  • Travel Award, Best Scientific Abstract, Association of University Anesthesiologists (3/2014)
  • T32 Training Grant in Anesthesia Research and Medicine, National Institutes of Health (7/2012-6/2014)
  • Fellow to Faculty Transition Award, 14FTF- 19970029, American Heart Association (7/2014 - 6/2019)

Boards, Advisory Committees, Professional Organizations

  • Member, American Heart Association (2014 - Present)
  • Member, International Anesthesia Research Society (2014 - Present)
  • Member, Society for Neuroscience (2013 - Present)
  • Member, Society for Neuroscience in Anesthesiology and Critical Care (2012 - Present)
  • Member, American Society of Anesthesiologists (2009 - Present)
  • Member, American College of Sports Medicine (1995 - Present)
  • Member, American Physiological Society (1994 - Present)

Professional Education

  • Board Certification: Anesthesia, American Board of Anesthesiology (2014)
  • Board Certification, American Board of Anesthesiology (2014)
  • Residency:UCSD School of Medicine (2012) CA
  • Medical Education:UCSD School of Medicine (2008) CA
  • Internship:University Of Hawaii (2009) HI
  • PhD, UC San Diego, Biomedicine (2006)
  • BS, UC San Diego, Animal Physiology and Neuroscience (1997)
  • BA, UC San Diego, Psychology (1997)

Research & Scholarship

Current Research and Scholarly Interests

I am interested in finding new strategies to promote neuronal survival and improve functional outcome following injury to the brain. The brain consists of several different cell types, the most abundant of which are astrocytes, specialized glial cells that play a vital role in regulating neuronal function and homeostasis. My current area of focus is utilizing microRNA's to simultaneously target multiple pathways which regulate cell survival in both neurons and astrocytes in order to minimize the degree of cell death subsequent to noxious stimuli, such as ischemia-reperfusion injury.


All Publications

  • Epigenetics: The Epicenter for Future Anesthesia Research? Anesthesiology Stary, C. M., Patel, H. H., Roth, D. M. 2015; 123 (4): 743-744

    View details for DOI 10.1097/ALN.0000000000000808

    View details for PubMedID 26259141

  • Astrocytes Protect against Isoflurane Neurotoxicity by Buffering pro-brain-derived Neurotrophic Factor. Anesthesiology Stary, C. M., Sun, X., Giffard, R. G. 2015; 123 (4): 810-819


    Isoflurane induces cell death in neurons undergoing synaptogenesis via increased production of pro-brain-derived neurotrophic factor (proBDNF) and activation of postsynaptic p75 neurotrophin receptor (p75). Astrocytes express p75, but their role in neuronal p75-mediated cell death remains unclear. The authors investigated whether astrocytes have the capacity to buffer increases in proBDNF and protect against isoflurane/p75 neurotoxicity.Cell death was assessed in day in vitro (DIV) 7 mouse primary neuronal cultures alone or in co-culture with age-matched or DIV 21 astrocytes with propidium iodide 24 h after 1 h exposure to 2% isoflurane or recombinant proBDNF. Astrocyte-targeted knockdown of p75 in co-culture was achieved with small-interfering RNA and astrocyte-specific transfection reagent and verified with immunofluorescence microscopy. proBDNF levels were assessed by enzyme-linked immunosorbent assay. Each experiment used six to eight replicate cultures/condition and was repeated at least three times.Exposure to isoflurane significantly (P < 0.05) increased neuronal cell death in primary neuronal cultures (1.5 ± 0.7 fold, mean ± SD) but not in co-culture with DIV 7 (1.0 ± 0.5 fold) or DIV 21 astrocytes (1.2 ± 1.2 fold). Exogenous proBDNF dose dependently induced neuronal cell death in both primary neuronal and co-cultures, an effect enhanced by astrocyte p75 inhibition. Astrocyte-targeted p75 knockdown in co-cultures increased media proBDNF (1.2 ± 0.1 fold) and augmented isoflurane-induced neuronal cell death (3.8 ± 3.1 fold).The presence of astrocytes provides protection to growing neurons by buffering increased levels of proBDNF induced by isoflurane. These findings may hold clinical significance for the neonatal and injured brain where increased levels of proBDNF impair neurogenesis.

    View details for DOI 10.1097/ALN.0000000000000824

    View details for PubMedID 26270940

  • Advances in Astrocyte-targeted Approaches for Stroke Therapy: An Emerging Role for Mitochondria and microRNAS NEUROCHEMICAL RESEARCH Stary, C. M., Giffard, R. G. 2015; 40 (2): 301-307


    Astrocytes are critical regulators of neuronal function and an effective target for stroke therapy in animal models. Identifying individual targets with the potential for simultaneous activation of multiple downstream pathways that regulate astrocyte homeostasis may be a necessary element for successful clinical translation. Mitochondria and microRNAs each represent individual targets with multi-modal therapeutic potential. Mitochondria regulate metabolism and apoptosis, while microRNAs have the capacity to bind and inhibit numerous mRNAs. By combining strategies targeted at maintaining astrocyte function during and following cerebral ischemia, a synergistic therapeutic effect may be achieved.

    View details for DOI 10.1007/s11064-014-1373-4

    View details for Web of Science ID 000349766200009

  • MicroRNA-200c Contributes to Injury From Transient Focal Cerebral Ischemia by Targeting Reelin STROKE Stary, C. M., Xu, L., Sun, X., Ouyang, Y., White, R. E., Leong, J., Li, J., Xiong, X., Giffard, R. G. 2015; 46 (2): 551-U356
  • MicroRNA-200c contributes to injury from transient focal cerebral ischemia by targeting Reelin. Stroke; a journal of cerebral circulation Stary, C. M., Xu, L., Sun, X., Ouyang, Y., White, R. E., Leong, J., Li, J., Xiong, X., Giffard, R. G. 2015; 46 (2): 551-556


    MicroRNA (miR)-200c increases rapidly in the brain after transient cerebral ischemia but its role in poststroke brain injury is unclear. Reelin, a regulator of neuronal migration and synaptogenesis, is a predicted target of miR-200c. We hypothesized that miR-200c contributes to injury from transient cerebral ischemia by targeting reelin.Brain infarct volume, neurological score and levels of miR-200c, reelin mRNA, and reelin protein were assessed in mice subjected to 1 hour of middle cerebral artery occlusion with or without intracerebroventricular infusion of miR-200c antagomir, mimic, or mismatch control. Direct targeting of reelin by miR-200c was assessed in vitro by dual luciferase assay and immunoblot.Pretreatment with miR-200c antagomir decreased post-middle cerebral artery occlusion brain levels of miR-200c, resulting in a significant reduction in infarct volume and neurological deficit. Changes in brain levels of miR-200c inversely correlated with reelin protein expression. Direct targeting of the Reln 3' untranslated region by miR-200c was verified with dual luciferase assay. Inhibition of miR-200c resulted in an increase in cell survival subsequent to in vitro oxidative injury. This effect was blocked by knockdown of reelin mRNA, whereas application of reelin protein afforded protection.These findings suggest that the poststroke increase in miR-200c contributes to brain cell death by inhibiting reelin expression, and that reducing poststroke miR-200c is a potential target to mitigate stroke-induced brain injury.

    View details for DOI 10.1161/STROKEAHA.114.007041

    View details for PubMedID 25604249

  • Caveolins: targeting pro-survival signaling in the heart and brain. Frontiers in physiology Stary, C. M., Tsutsumi, Y. M., Patel, P. M., Head, B. P., Patel, H. H., Roth, D. M. 2012; 3: 393-?


    The present review discusses intracellular signaling moieties specific to membrane lipid rafts (MLRs) and the scaffolding proteins caveolin and introduces current data promoting their potential role in the treatment of pathologies of the heart and brain. MLRs are discreet microdomains of the plasma membrane enriched in gylcosphingolipids and cholesterol that concentrate and localize signaling molecules. Caveolin proteins are necessary for the formation of MLRs, and are responsible for coordinating signaling events by scaffolding and enriching numerous signaling moieties in close proximity. Specifically in the heart and brain, caveolins are necessary for the cytoprotective phenomenon termed ischemic and anesthetic preconditioning. Targeted overexpression of caveolin in the heart and brain leads to induction of multiple pro-survival and pro-growth signaling pathways; thus, caveolins represent a potential novel therapeutic target for cardiac and neurological pathologies.

    View details for DOI 10.3389/fphys.2012.00393

    View details for PubMedID 23060817

  • Physiologically normal 5% O2 supports neuronal differentiation and resistance to inflammatory injury in neural stem cell cultures. Journal of neuroscience research Sun, X., Voloboueva, L. A., Stary, C. M., Giffard, R. G. 2015; 93 (11): 1703-1712


    Recent studies have demonstrated that neural stem cell (NSC) culture at physiologically normoxic conditions (2-5% O2 ) is advantageous in terms of neuronal differentiation and survival. Neuronal differentiation is accompanied by a remarkable shift to mitochondrial oxidative metabolism compared with preferentially glycolytic metabolism of proliferating cells. However, metabolic changes induced by growth in a normoxic (5%) O2 culture environment in NSCs have been minimally explored. This study demonstrates that culturing under 5% O2 conditions results in higher levels of mitochondrial oxidative metabolism, decreased glycolysis, and reduced levels of reactive oxygen species in NSC cultures. Inflammation is one of the major environmental factors limiting postinjury NSC neuronal differentiation and survival. Our results show that NSCs differentiated under 5% O2 conditions possess better resistance to in vitro inflammatory injury compared with those exposed to 20% O2 . The present work demonstrates that lower, more physiologically normal O2 levels support metabolic changes induced during NSC neuronal differentiation and provide increased resistance to inflammatory injury, thus highlighting O2 tension as an important determinant of cell fate and survival in various stem cell therapies. © 2015 Wiley Periodicals, Inc.

    View details for DOI 10.1002/jnr.23615

    View details for PubMedID 26147710

  • T Cells and Cerebral Ischemic Stroke NEUROCHEMICAL RESEARCH Gu, L., Jian, Z., Stary, C., Xiong, X. 2015; 40 (9): 1786-1791
  • Post-stroke treatment with miR-181 antagomir reduces injury and improves long-term behavioral recovery in mice after focal cerebral ischemia EXPERIMENTAL NEUROLOGY Xu, L., Ouyang, Y., Xiong, X., Stary, C. M., Giffard, R. G. 2015; 264: 1-7


    miR-181 has deleterious effects on stroke outcome, and reducing miR-181a levels prior to middle cerebral artery occlusion (MCAO) was shown previously to be protective. Here we tested the effect of post-ischemic treatment with miR-181a antagomir by intracerebroventricular and intravenous routes of administration on infarct size, neurological outcome, inflammatory response and long term behavioral outcome. Post-treatment with miR-181a antagomir significantly reduced infarction size, improved neurological deficits and reduced NF-κB activation, numbers of infiltrating leukocytes and levels of Iba1. Targets affected by miR-181a antagomir administered after stroke onset include BCL2 and X-linked inhibitor of apoptosis protein (XIAP). Post-treatment with miR-181a antagomir significantly improved behavioral outcome assessed by rotarod at one month. These findings indicate that post-treatment with miR-181a antagomir has neuroprotective effects against ischemic neuronal damage and neurological impairment in mice, and the protection is long lasting including recovery of motor function and coordination over one month. The ability to protect the brain with post-treatment with miR-181a antagomir with long lasting effect makes this a promising therapeutic target and may be an innovative and effective new approach for stroke therapy.

    View details for DOI 10.1016/j.expneurol.2014.11.007

    View details for Web of Science ID 000349590400001

  • Role of caveolin-3 in lymphocyte activation LIFE SCIENCES Tran, C., Stary, C. M., Schilling, J. M., Bentley, B., Patel, H. H., Roth, D. M. 2015; 121: 35-39


    Caveolins are structural proteins clustered in lipid-rich regions of plasma membrane involved in coordinating signal transduction in various organ systems. While caveolin-1 (Cav-1) has been shown to regulate lymphocyte activation, the role of caveolin-3 (Cav-3) in immune system signaling has not been investigated. We tested the hypothesis that Cav-3 modulates lymphocyte activation.Lymphocyte/leukocyte subpopulations from WT and Cav-3 mice were profiled with flow cytometry. Cytokine production in quiescent and activated splenocytes from WT and Cav-3 mice was assessed with ELISA.Levels of T-cells, monocytes, and natural killer cells were not different between WT and KO mice, however KO mice had lower B-cell population-percentage. Functionally, activated lymphocytes from Cav-3 KO mice demonstrated significantly reduced expression of IL-2 compared to WT, while expression of TNFα, IL-6, and IL-10 was not different. Finally, expression of IL-17 was significantly reduced in T-helper cells from KO mice, while IFNγ was not, suggesting that Cav-3 is a determinant in the development of the Th-17 subpopulation.This study is the first to demonstrate that Cav-3 may be a novel participant in B-cell expression, T-cell cytokine production and activation of inflammation.

    View details for DOI 10.1016/j.lfs.2014.11.017

    View details for Web of Science ID 000349590800006

  • The Use of microRNAs to Modulate Redox and Immune Response to Stroke ANTIOXIDANTS & REDOX SIGNALING Ouyang, Y., Stary, C. M., White, R. E., Giffard, R. G. 2015; 22 (2): 187-202


    Significance: Cerebral ischemia is a major cause of death and disability throughout the world, yet therapeutic options remain limited. The interplay between cellular redox state and the immune response plays a critical role in determining the extent of neural cell injury after ischemia and reperfusion. Excessive amounts of reactive oxygen species (ROS) generated by mitochondria and other sources act both as triggers and effectors of inflammation. This review will focus on the interplay between these two mechanisms. Recent Advances: MicroRNAs (miRNAs) are important post-transcriptional regulators that interact with multiple target messenger RNAs (mRNAs) coordinately regulating target genes including those involved in controlling mitochondrial function, redox state, and inflammatory pathways. This review will focus on the regulation of mitochondria, ROS, and inflammation by miRNAs in the chain of deleterious intra- and intercellular events that lead to brain cell death after cerebral ischemia. Critical Issues: Although pretreatment using miRNAs was effective in cerebral ischemia in rodents, testing treatment after the onset of ischemia is an essential next step in the development of acute stroke treatment. In addition, miRNA formulation and delivery into the CNS remain a challenge in the clinical translation of miRNA therapy. Future Directions: Future research should focus on post-treatment and potential clinical use of miRNAs.

    View details for DOI 10.1089/ars.2013.5757

    View details for Web of Science ID 000347037800008

  • Molecular Pathogenesis of Anti-NMDAR Encephalitis. BioMed research international Ding, H., Jian, Z., Stary, C. M., Yi, W., Xiong, X. 2015; 2015: 643409-?


    Anti-NMDAR encephalitis is a recently identified autoimmune disease, described by an immune-mediated loss of NMDA glutamate receptors, resulting in progressive mental deterioration. To date, literature on anti-NMDAR encephalitis has been largely clinically oriented, including descriptions of the clinical presentation and course, diagnostic methods, and potential clinical treatments. However, the underlying molecular mechanisms contributing to the complex immunological cellular transformation that is associated with the progression of anti-NMDAR encephalitis remain to be adequately explored. This review will provide a summary of the current literature on anti-NMDAR encephalitis, including the immunologic molecular mechanisms contributing to disease progression. In particular this review will focus on the effect of anti-NMDAR on GluN2-NMDAR expression and the molecular transformation of B and T leukocytes in the loss of self-tolerance. Further research on the immunologic mechanisms contributing to anti-NMDAR encephalitis may provide an avenue for future novel diagnostic approaches, such as immunologic surveillance, as well as new therapeutic strategies for this recently identified autoimmune disease.

    View details for DOI 10.1155/2015/643409

    View details for PubMedID 26221602

  • microRNAs: Innovative Targets for Cerebral Ischemia and Stroke CURRENT DRUG TARGETS Ouyang, Y., Stary, C. M., Yang, G., Giffard, R. 2013; 14 (1): 90-101


    Stroke is one of the leading causes of death and disability worldwide. Because stroke is a multifactorial disease with a short therapeutic window many clinical stroke trials have failed and the only currently approved therapy is thrombolysis. MicroRNAs (miRNA) are endogenously expressed noncoding short single-stranded RNAs that play a role in the regulation of gene expression at the post-transcriptional level, via degradation or translational inhibition of their target mRNAs. The study of miRNAs is rapidly growing and recent studies have revealed a significant role of miRNAs in ischemic disease. miRNAs are especially important candidates for stroke therapeutics because of their ability to simultaneously regulate many target genes and since to date targeting single genes for therapeutic intervention has not yet succeeded in the clinic. Although there are already quite a few review articles about miRNA in ischemic heart disease, much less is currently known about miRNAs in cerebral ischemia. This review summarizes current knowledge about miRNAs and cerebral ischemia, focusing on the role of miRNAs in ischemia, both changes in expression and identification of potential targets, as well as the potential of miRNAs as biomarkers and therapeutic targets in cerebral ischemia.

    View details for Web of Science ID 000316871200010

  • Idiopathic granulomatous mastitis associated with corynebacterium sp. Infection. Hawaii medical journal Stary, C. M., Lee, Y. S., Balfour, J. 2011; 70 (5): 99-101


    Idiopathic granulomatous mastitis (IGM) is a rare inflammatory condition of the breast. The etiology and treatments options of IGM remain controversial. Previous case reports have suggested that Corynebacterium sp., a gram-positive bacillus endogenous to the skin, may be associated with IGM. In the present report, we describe the first case of IGM with a positive culture for Corynebacterium sp. reported in the United States.

    View details for PubMedID 21857740

  • The O-2 cost of the tension-time integral in isolated single myocytes during fatigue AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY Hepple, R. T., Howlett, R. A., Kindig, C. A., Stary, C. M., Hogan, M. C. 2010; 298 (4): R983-R988


    One proposed explanation for the Vo(2) slow component is that lower-threshold motor units may fatigue and develop little or no tension but continue to use O(2), thereby resulting in a dissociation of cellular respiration from force generation. The present study used intact isolated single myocytes with differing fatigue resistance profiles to investigate the relationship between fatigue, tension development, and aerobic metabolism. Single Xenopus skeletal muscle myofibers were allocated to a fast-fatiguing (FF) or a slow-fatiguing (SF) group, based on the contraction frequency required to elicit a fall in tension to 60% of peak. Phosphorescence quenching of a porphyrin compound was used to determine Delta intracellular Po(2) (Pi(O(2)); a proxy for Vo(2)), and developed isometric tension was monitored to allow calculation of the time-integrated tension (TxT). Although peak DeltaPi(O(2)) was not different between groups (P = 0.36), peak tension was lower (P < 0.05) in SF vs. FF (1.97 +/- 0. 17 V vs. 2. 73 +/- 0.30 V, respectively) and time to 60% of peak tension was significantly longer in SF vs. FF (242 +/- 10 s vs. 203 +/- 10 s, respectively). Before fatigue, both DeltaPi(O(2)) and TxT rose proportionally with contraction frequency in SF and FF, resulting in DeltaPi(O(2))/TxT being identical between groups. At fatigue, TxT fell dramatically in both groups, but DeltaPi(O(2)) decreased proportionately only in the FF group, resulting in an increase in DeltaPi(O(2))/TxT in the SF group relative to the prefatigue condition. These data show that more fatigue-resistant fibers maintain aerobic metabolism as they fatigue, resulting in an increased O(2) cost of contractions that could contribute to the Vo(2) slow component seen in whole body exercise.

    View details for DOI 10.1152/ajpregu.00715.2009

    View details for Web of Science ID 000275857800014

    View details for PubMedID 20130224

  • Glycolytic activation at the onset of contractions in isolated Xenopus laevis single myofibres EXPERIMENTAL PHYSIOLOGY Walsh, B., Stary, C. M., Howlett, R. A., Kelley, K. M., Hogan, M. C. 2008; 93 (9): 1076-1084


    Intracellular pH (pHi) was measured in isolated Xenopus laevis single myofibres at the onset of contractions, with and without glycolytic blockade, to investigate the time course of glycolytic activation. Single myofibres (n=8; CON) were incubated in 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein acetoyxmethyl ester (10 microM; for fluorescence measurement of pHi) and stimulated for 15 s at 0.67 Hz in anoxia in the absence (control condition; CON) and presence of a glycolytic inhibitor (1 mM iodoacetic acid; IAA). Intracellular pHi and tension were continuously recorded, and the differences in pHi between conditions were used to estimate the activation time of glycolysis. An immediate and steady increase in pHi (initial alkalosis) at the onset of contractions was similar between CON and IAA trials for the first 9 s of the contractile bout. However, from six contractions (approximately 10 s) throughout the remainder of the bout, IAA demonstrated a continued rise in pHi, in contrast to a progressive decrease in pHi in CON (P<0.05). These results demonstrate, with high temporal resolution, that glycolysis is activated within six contractions (10 s at 0.67 Hz) in single Xenopus skeletal muscle fibres.

    View details for DOI 10.1113/expphysiol.2008.042440

    View details for Web of Science ID 000258442700008

    View details for PubMedID 18515473

  • Elevation in heat shock protein 72 mRNA following contractions in isolated single skeletal muscle fibers AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY Stary, C. M., Walsh, B. J., Knapp, A. E., Brafman, D., Hogan, M. C. 2008; 295 (2): R642-R648


    The purpose of the present study was 1) to develop a stable model for measuring contraction-induced elevations in mRNA in single skeletal muscle fibers and 2) to utilize this model to investigate the response of heat shock protein 72 (HSP72) mRNA following an acute bout of fatiguing contractions. Living, intact skeletal muscle fibers were microdissected from lumbrical muscle of Xenopus laevis and either electrically stimulated for 15 min of tetanic contractions (EX; n=26) or not stimulated to contract (REST; n=14). The relative mean developed tension of EX fibers decreased to 29+/-7% of initial peak tension at the stimulation end point. Following treatment, individual fibers were allowed to recover for 1 (n=9), 2 (n=8), or 4 h (n=9) prior to isolation of total cellular mRNA. HSP72, HSP60, and cardiac alpha-actin mRNA content were then assessed in individual fibers using quantitative PCR detection. Relative HSP72 mRNA content was significantly (P<0.05) elevated at the 2-h postcontraction time point relative to REST fibers when normalized to either HSP60 (18.5+/-7.5-fold) or cardiac alpha-actin (14.7+/-4.3-fold), although not at the 1- or 4-h time points. These data indicate that 1) extraction of RNA followed by relative quantification of mRNA of select genes in isolated single skeletal muscle fibers can be reliably performed, 2) HSP60 and cardiac alpha-actin are suitable endogenous normalizing genes in skeletal muscle following contractions, and 3) a significantly elevated content of HSP72 mRNA is detectable in skeletal muscle 2 h after a single bout of fatiguing contractions, despite minimal temperature changes and without influence from extracellular sources.

    View details for DOI 10.1152/ajpregu.00852.2007

    View details for Web of Science ID 000258322700032

    View details for PubMedID 18525012

  • Measurement of activation energy and oxidative phosphorylation onset kinetics in isolated muscle fibers in the absence of cross-bridge cycling AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY Walsh, B., Howlett, R. A., Stary, C. M., Kindig, C. A., Hogan, M. C. 2006; 290 (6): R1707-R1713


    This study utilized N-benzyl-p-toluene sulfonamide (BTS), a potent inhibitor of cross-bridge cycling, to measure 1) the relative metabolic costs of cross-bridge cycling and activation energy during contraction, and 2) oxygen uptake kinetics in the presence and absence of myosin ATPase activity, in isolated Xenopus laevis muscle fibers. Isometric tension development and either cytosolic Ca2+ concentration ([Ca2+]c) or intracellular Po2 (PiO2) were measured during contractions at 20 degrees C in control conditions (Con) and after exposure to 12.5 microM BTS. BTS attenuated tension development to 5+/-0.4% of Con but did not affect either resting or peak [Ca2+]c during repeated isometric contractions. To determine the relative metabolic cost of cross-bridge cycling, we measured the fall in PiO2) (DeltaPiO2; a proxy for Vo2) during contractions in Con and BTS groups. BTS attenuated DeltaP(iO2) by 55+/-6%, reflecting the relative ATP cost of cross-bridge cycling. Thus, extrapolating DeltaPiO2 to a value that would occur at 0% tension suggests that actomyosin ATP requirement is approximately 58% of overall ATP consumption during isometric contractions in mixed fiber types. BTS also slowed the fall in PiO2) (time to 63% of overall DeltaPiO2) from 75+/-9 s (Con) to 101+/-9 s (BTS) (P<0.05), suggesting an important role of the products of ATP hydrolysis in determining the Vo2 onset kinetics. These results demonstrate in isolated skeletal muscle fibers that 1) activation energy accounts for a substantial proportion (approximately 42%) of total ATP cost during isometric contractions, and 2) despite unchanged [Ca2+]c transients, a reduced rate of ATP consumption results in slower Vo2 onset kinetics.

    View details for DOI 10.1152/ajpregu.00687.2005

    View details for Web of Science ID 000237368500031

    View details for PubMedID 16424084

  • Determinants of oxidative phosphorylation onset kinetics in isolated myocytes MEDICINE AND SCIENCE IN SPORTS AND EXERCISE Walsh, B., Howlett, R. A., Stary, C. M., Kindig, C. A., Hogan, M. C. 2005; 37 (9): 1551-1558


    At the onset of constant-load exercise, pulmonary oxygen uptake (VO(2)) exhibits a monoexponential increase, following a brief time delay, to a new steady state. To date, the specific factors controlling VO(2) onset kinetics during the transition to higher rates of work remain largely unknown. To study the control of respiration in the absence of confounding factors such as blood flow heterogeneity and fiber type recruitment patterns, the onset kinetics of mitochondrial respiration were studied at the start of contractions in isolated single myocytes. Individual myocytes were microinjected with a porphyrin compound to allow phosphorescent measurement of intracellular PO(2) (P(i)O(2), an analog of VO(2)). Peak tension and P(i)O(2) were continuously monitored under a variety of conditions designed to test the role of work intensity, extracellular PO(2), cellular metabolites, and enzyme activation on the regulation of VO(2) onset kinetics.

    View details for DOI 10.1249/01.mss.0000177469.25763.25

    View details for Web of Science ID 000232078100015

    View details for PubMedID 16177608

  • Intracellular pH during sequential, fatiguing contractile periods in isolated single Xenopus skeletal muscle fibers JOURNAL OF APPLIED PHYSIOLOGY Stary, C. M., Hogan, M. C. 2005; 99 (1): 308-312


    The purpose of the present study was to test the hypothesis that a preceding contractile period in isolated single skeletal muscle fibers would attenuate the decrease in pH during an identical, subsequent contractile period, thereby reducing the rate of fatigue. Intact single skeletal muscle fibers (n = 9) were isolated from Xenopus lumbrical muscle and incubated with the fluorescent cytosolic H+ indicator 2',7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) AM for 30 min. Two identical contractile periods were performed in each fiber, separated by a 1-h recovery period. Force and intracellular pH (pHi) fluorescence were measured simultaneously while fibers were stimulated (tetanic contractions of 350-ms trains with 70-Hz stimuli at 9 V) at progressively increasing frequencies (0.25, 0.33, 0.5, and 1 contraction/s) until the development of fatigue (to 60% initial force). No significant difference (P < 0.05) was observed between the first and second contractile periods in initial force development, resting pHi, or time to fatigue (5.3 +/- 0.5 vs. 5.1 +/- 0.6 min). However, the relative decrease in the BCECF fluorescence ratio (and therefore pHi) from rest to the fatigue time point was significantly greater (P < 0.05) during the first contractile period (to 65 +/- 4% of initial resting values) compared with the second (77 +/- 4%). The results of the present study demonstrated that, when preceded by an initial fatiguing contractile period, the rise in cytosolic H+ concentration in contracting single skeletal muscle fibers during a second contractile period was significantly reduced but did not attenuate the fatigue process in the second contractile period. These results suggest that intracellular factors other than H+ accumulation contribute to the fall in force development under these conditions.

    View details for DOI 10.1152/japplphysiol.01361.2004

    View details for Web of Science ID 000230385800041

    View details for PubMedID 15761085

  • Relationship between intracellular PO2 recovery kinetics and fatigability in isolated single frog myocytes JOURNAL OF APPLIED PHYSIOLOGY Kindig, C. A., Walsh, B., Howlett, R. A., Stary, C. M., Hogan, M. C. 2005; 98 (6): 2316-2319


    In single frog skeletal myocytes, a linear relationship exists between "fatigability" and oxidative capacity. The purpose of this investigation was to study the relationship between the intracellular Po(2) (Pi(O(2))) offset kinetics and fatigability in single Xenopus laevis myocytes to test the hypothesis that Pi(O(2)) offset kinetics would be related linearly with myocyte fatigability and, by inference, oxidative capacity. Individual myocytes (n = 30) isolated from lumbrical muscle were subjected to a 2-min bout of isometric peak tetanic contractions at either 0.25- or 0.33-Hz frequency while Pi(O(2)) was measured continuously via phosphorescence quenching techniques. The mean response time (MRT; time to 63% of the overall response) for Pi(O(2)) recovery from contracting values to resting baseline was calculated. After the initial square-wave constant-frequency contraction trial, each cell performed an incremental contraction protocol [i.e., frequency increase every 2 min from 0.167, 0.25, 0.33, 0.5, 1.0, and 2.0 Hz until peak tension fell below 50% of initial values (TTF)]. TTF values ranged from 3.39 to 10.04 min for the myocytes. The Pi(O(2)) recovery MRT ranged from 26 to 146 s. A significant (P < 0.05), negative relationship (MRT = -12.68TTF + 168.3, r(2) = 0.605) between TTF and Pi(O(2)) recovery MRT existed. These data demonstrate a significant correlation between fatigability and oxidative phosphorylation recovery kinetics consistent with the notion that oxidative capacity determines, in part, the speed with which skeletal muscle can recover energetically to alterations in metabolic demand.

    View details for DOI 10.1152/japplphysiol.00355.2004

    View details for Web of Science ID 000229365500047

    View details for PubMedID 15691906

  • NAD(P)H fluorescence imaging of mitochondrial metabolism in contracting Xenopus skeletal muscle fibers: effect of oxygen availability JOURNAL OF APPLIED PHYSIOLOGY Hogan, M. C., Stary, C. M., Balaban, R. S., Combs, C. A. 2005; 98 (4): 1420-1426


    The blue autofluorescence (351 nm excitation, 450 nm emission) of single skeletal muscle fibers from Xenopus was characterized to be originating from mitochondrial NAD(P)H on the basis of morphological and functional correlations. This fluorescence signal was used to estimate the oxygen availability to isolated single Xenopus muscle fibers during work level transitions by confocal microscopy. Fibers were stimulated to generate two contractile periods that were only different in the PO2 of the solution perfusing the single fibers (PO2 of 30 or 0-2 Torr; pH = 7.2). During contractions, mean cellular NAD(P)H increased significantly from rest in the low PO2 condition with the core (inner 10%) increasing to a greater extent than the periphery (outer 10%). After the cessation of work, NAD(P)H decreased in a manner consistent with oxygen tensions sufficient to oxidize the surplus NAD(P)H. In contrast, NAD(P)H decreased significantly with work in 30 Torr PO2. However, the rate of NAD(P)H oxidation was slower and significantly increased with the cessation of work in the core of the fiber compared with the peripheral region, consistent with a remaining limitation in oxygen availability. These results suggest that the blue autofluorescence signal in Xenopus skeletal muscle fibers is from mitochondrial NAD(P)H and that the rate of NAD(P)H oxidation within the cell is influenced by extracellular PO2 even at high extracellular PO2 during the contraction cycle. These results also demonstrate that although oxygen availability influences the rate of NAD(P)H oxidation, it does not prevent NAD(P)H from being oxidized through the process of oxidative phosphorylation at the onset of contractions.

    View details for DOI 10.1152/japplphysiol.00849.2004

    View details for Web of Science ID 000227665400037

    View details for PubMedID 15591295

  • Effects of acute creatine kinase inhibition on metabolism and tension development in isolated single myocytes JOURNAL OF APPLIED PHYSIOLOGY Kindig, C. A., Howlett, R. A., Stary, C. M., Walsh, B., Hogan, M. C. 2005; 98 (2): 541-549


    This study investigated the effects of acute creatine kinase (CK) inhibition (CKi) on contractile performance, cytosolic Ca2+ concentration ([Ca2+]c), and intracellular PO2 (PIO2) in Xenopus laevis isolated myocytes during a 2-min bout of isometric tetanic contractions (0.33-Hz frequency). Peak tension was similar between trials during the first contraction but was significantly (P < 0.05) attenuated for all subsequent contractions in CKi vs. control (Con). The fall in PIO2 (DeltaPIO2) from resting values was significantly greater in Con (26.0 +/- 2.2 Torr) compared with CKi (17.8 +/- 1.8 Torr). However, the ratios of Con to CKi end-peak tension (1.53 +/- 0.11) and DeltaPO2 (1.49 +/- 0.11) were similar, suggesting an unaltered aerobic cost of contractions. Additionally, the mean response time (MRT) of DeltaPIO2was significantly faster in CKi vs. Con during both the onset (31.8 +/- 5.5 vs. 49.3 +/- 5.7 s; P < 0.05) and cessation (21.2 +/- 4.1 vs. 68.0 +/- 3.2 s; P < 0.001) of contractions. These data demonstrate that initial phosphocreatine hydrolysis in single skeletal muscle fibers is crucial for maintenance of sarcoplasmic reticulum Ca2+ release and peak tension during a bout of repetitive tetanic contractions. Furthermore, as PIO2 fell more rapidly at contraction onset in CKi compared with Con, these data suggest that CK activity temporally buffers the initial ATP-to-ADP concentration ratio at the transition to an augmented energetic demand, thereby slowing the initial mitochondrial activation by mitigating the energetic control signal (i.e., ADP concentration, phosphorylation potential, etc.) between sites of ATP supply and demand.

    View details for DOI 10.1152/japplphysiol.00354.2004

    View details for Web of Science ID 000226282400017

    View details for PubMedID 15333609

  • Effect of dissociating cytosolic calcium and metabolic rate of intracellular Po-2 kinetics in single frog myocytes JOURNAL OF PHYSIOLOGY-LONDON Kindig, C. A., Stary, C. M., Hogan, M. C. 2005; 562 (2): 527-534


    The purpose of this investigation was to utilize 2,3-butanedione monoxime (BDM; an inhibitor of contractile activation) to dissociate cytosolic [Ca(2+)] ([Ca(2+)](c)) from the putative respiratory regulators that arise from muscle contraction-induced ATP utilization in order to determine the relative contribution of [Ca(2+)](c) on intracellular P(O(2)) (P(iO(2))) kinetics during the transition from rest to contractions in single skeletal myocytes isolated from Xenopus laevis lumbrical muscle. Myocytes were subjected to electrically induced isometric tetanic contractions (0.25 Hz; 2-min bouts) while peak tension and either [Ca(2+)](c) (n= 7; ratiometric fluorescence microscopy) or P(iO(2)) (n= 7; phosphorescence microscopy) was measured continuously. Cells were studied under both control and 3 mm BDM conditions in randomized order. Initial (control, 100 +/- 0%; BDM, 72.6 +/- 4.6%), midpoint (control, 86.7 +/- 1.8%; BDM, 61.6 +/- 4.1%) and end (control, 85.0 +/- 2.8%; BDM, 57.5 +/- 5.0%) peak tensions (normalized to initial control values) were significantly reduced (P < 0.05) with BDM compared with control (n= 14). Despite the reduced peak tension, peak [Ca(2+)](c) was not altered (P > 0.05) between control and BDM trials. Thus, the peak tension-to-peak [Ca(2+)](c) ratio was reduced with BDM compared with control. The absolute fall in P(iO(2)) with contractions, which is proportional to the rise in , was significantly reduced with BDM (13.2 +/- 1.3 mmHg) compared with control (22.0 +/- 2.0 mmHg). However, P(iO(2)) onset kinetics (i.e. mean response time (MRT)) was not altered between BDM (66.8 +/- 8.0 s) and control (64.9 +/- 6.3 s) trials. Therefore, the initial rate of change (defined as the fall in P(iO(2))/MRT) was significantly slower in BDM fibres compared with control. These data demonstrate in these isolated single skeletal muscle fibres that unchanged peak [Ca(2+)](c) in the face of reduced metabolic feedback from the contractile sites evoked with BDM did not alter P(iO(2)) onset kinetics in isolated single frog myocytes, suggesting that metabolic signals arising from the contractile sites play a more substantial role than [Ca(2+)](c) in the signalling pathway to oxidative phosphorylation during the transition from rest to repeated tetanic contractions.

    View details for DOI 10.1113/jphysiol.2004.074922

    View details for Web of Science ID 000226528000016

    View details for PubMedID 15550463

  • Resistance to fatigue of individual Xenopus single skeletal muscle fibres is correlated with mitochondrial volume density EXPERIMENTAL PHYSIOLOGY Stary, C. M., Mathieu-Costello, O., Hogan, M. C. 2004; 89 (5): 617-621


    The purpose of the present study was to compare the individual fatigue characteristics of isolated single skeletal muscle fibres with their mitochondrial volume density (MVD), using direct histological morphometry. Single muscle fibres (n= 14) were microdissected from lumbrical muscle of adult female Xenopus laevis, and force was measured while fibres were stimulated (tetanic contractions of 200 ms trains with 70 Hz stimuli at 9 V) at progressively increasing frequencies (2 min each at 0.25, 0.33, 0.5 and 1 contractions s(-1)) until fatigue (<50% initial maximal force) had been established. Following the end of the fatigue protocol, MVD was determined by electron microscopy. Time to fatigue varied among the individual fibres from 3.3 to 10 min. MVD of individual fibres ranged from 3.0 to 9.2% and was positively correlated (r= 0.93) with time to fatigue of corresponding fibres. These results, using direct histological measurements of MVD: (1) support on a single cell basis the notion that oxidative capacity is a major determinant of muscle fatigue resistance; and (2) show that the fatigue profile of a single cell can be used to predict oxidative capacity.

    View details for DOI 10.1113/expphysiol.2004.027763

    View details for Web of Science ID 000223470200012

    View details for PubMedID 15258122

  • Effect of contraction frequency on the contractile and noncontractile phases of muscle venous blood flow JOURNAL OF APPLIED PHYSIOLOGY Hogan, M. C., Grassi, B., Samaja, M., Stary, C. M., Gladden, L. B. 2003; 95 (3): 1139-1144


    The purpose of this study was to test the hypothesis that increasing muscle contraction frequency, which alters the duty cycle and metabolic rate, would increase the contribution of the contractile phase to mean venous blood flow in isolated skeletal muscle during rhythmic contractions. Canine gastrocnemius muscle (n = 5) was isolated, and 3-min stimulation periods of isometric, tetanic contractions were elicited sequentially at rates of 0.25, 0.33, and 0.5 contractions/s. The O2 uptake, tension-time integral, and mean venous blood flow increased significantly (P < 0.05) with each contraction frequency. Venous blood flow during both the contractile (106 +/- 6, 139 +/- 8, and 145 +/- 8 ml x 100 g-1 x min-1) and noncontractile phases (64 +/- 3, 78 +/- 4, and 91 +/- 5 ml x 100 g-1 x min-1) increased with contraction frequency. Although developed force and duration of the contractile phase were never significantly different for a single contraction during the three contraction frequencies, the amount of blood expelled from the muscle during an individual contraction increased significantly with contraction frequency (0.24 +/- 0.03, 0.32 +/- 0.02, and 0.36 +/- 0.03 ml x N-1 x min-1, respectively). This increased blood expulsion per contraction, coupled with the decreased time in the noncontractile phase as contraction frequency increased, resulted in the contractile phase contribution to mean venous blood flow becoming significantly greater (21 +/- 4, 30 +/- 4, and 38 +/- 6%) as contraction frequency increased. These results demonstrate that the percent contribution of the muscle contractile phase to mean venous blood flow becomes significantly greater as contraction frequency (and thereby duty cycle and metabolic rate) increases and that this is in part due to increased blood expulsion per contraction.

    View details for DOI 10.1152/japplphysiol.00226.2003

    View details for Web of Science ID 000184733800031

    View details for PubMedID 12794032

  • Trimetazidine reduces basal cytosolic Ca2+ concentration during hypoxia in single Xenopus skeletal myocytes EXPERIMENTAL PHYSIOLOGY Stary, C. M., Kohin, S., Samaja, M., Howlett, R. A., Hogan, M. C. 2003; 88 (3): 415-421


    We tested the hypotheses that: (1) Ca(2+) handling and force production would be irreversibly altered in skeletal muscle during steady-state contractions when subjected to severe, prolonged hypoxia and subsequent reoxygenation; and (2) application of the cardio-protective drug trimetazidine would attenuate these alterations. Single, living skeletal muscle fibres from Xenopus laevis were injected with the Ca(2+) indicator fura 2, and incubated for 1 h prior to stimulation in 100 micro M TMZ-Ringer solution (TMZ; n = 6) or standard Ringer solution (CON; n = 6). Force and relative free cytosolic Ca(2+) concentration ([Ca(2+)](c)) were measured during continuous tetanic contractions produced every 5 s as fibres were sequentially perfused in the following manner: 3 min high extracellular P(O(2)) (159 mmHg), 15 min hypoxic perfusion (3-5 mmHg) then 3 min high P(O(2)). Hypoxia caused a decrease in force and peak [Ca(2+)](c) in both the TMZ and CON fibres, with no significant (P < 0.05) difference between groups. However, basal [Ca(2+)](c) was significantly lower during hypoxia in the TMZ group vs. the CON group. While reoxygenation generated only modest recovery of relative force and peak [Ca(2+)](c) in both groups, basal [Ca(2+)](c) remained significantly less in the TMZ group. These results demonstrated that in contracting, single skeletal muscle fibres, TMZ prevented increases in basal [Ca(2+)](c) generated during a severe hypoxic insult and subsequent reoxygenation, yet failed to protect the cell from the deleterious effects of prolonged hypoxia followed by reoxygenation.

    View details for Web of Science ID 000183476900013

    View details for PubMedID 12719766

  • No effect of trans sodium crocetinate on maximal O-2 conductance or V-O2,V-max in moderate hypoxia RESPIRATORY PHYSIOLOGY & NEUROBIOLOGY Hepple, R. T., Stary, C. M., Kohin, S., Wagner, P. D., Hogan, M. C. 2003; 134 (3): 239-246


    The lumped parameter describing skeletal muscle diffusional conductance for O(2), DM(O(2)), reflects all of the resistances for O(2) in moving from red cell to muscle fiber mitochondria. The purpose of our study was to determine if the carotenoid compound, trans sodium crocetinate (TSC), which has been reported to increase the diffusivity of O(2) in plasma, improves DM(O(2)) and thus, V(O(2),max) in maximally contracting in situ skeletal muscle. V(O(2),max) was measured in the isolated perfused canine gastrocnemius (n=5) during 3 min of isometric tetanic contractions at 1 Hz, while the animal was breathing 12% O(2) (PA(O(2))=32+/-2 Torr, mean+/-S.E.) under two experimental conditions. The first was a control contraction period and the second (following 60 min recovery) was performed within 5 min after infusion of a 0.1 mg x ml(-1) solution of TSC (total dose 100 microg kg(-1)). There were no significant differences in convective O(2) delivery (11.9+/-2.3 vs. 12.1+/-2.2 ml x min(-1) x 100 g(-1)), V(O(2),max) (9.5+/-1.5 vs. 9.6+/-1.5 ml x min(-1) x 100 g(-1)) or calculated DM(O(2)) (0.37+/-0.03 vs. 0.37+/-0.04 ml x min(-1) x 100 g(-1) x Torr(-1)) between contraction periods. As such, our results show that TSC does not improve performance in maximally contracting canine gastrocnemius muscle in situ under moderately hypoxic conditions, suggesting either that TSC in this situation does not increase plasma O(2) diffusivity or that this step in O(2) diffusion from red cell to myocyte does not constrain DM(O(2)).

    View details for Web of Science ID 000182101900006

    View details for PubMedID 12660103

  • Assessment of O-2 uptake dynamics in isolated single skeletal myocytes JOURNAL OF APPLIED PHYSIOLOGY Kindig, C. A., Kelley, K. M., Howlett, R. A., Stary, C. M., Hogan, M. C. 2003; 94 (1): 353-357


    The purpose of this research was to develop a technique for rapid measurement of O(2) uptake (Vo(2)) kinetics in single isolated skeletal muscle cells. Previous attempts to measure single cell Vo(2) have utilized polarographic-style electrodes, thereby mandating large fluid volumes and relatively poor sensitivity. Thus our laboratory has developed an approximately 100-microl, well-stirred chamber for the measurement of Vo(2) in isolated Xenopus laevis myocytes using a phosphorescence quenching technique [Ringer solution with 0.05 mM Pd-meso-tetra(4-carboxyphenyl)porphine] to monitor the fall in extracellular Po(2) (which is proportional to cellular Vo(2) within the sealed chamber). Vo(2) in single living myocytes dissected from Xenopus lumbrical muscles was measured from rest across a bout of repetitive tetanic contractions (0.33 Hz) and in response to a ramp protocol utilizing an increasing contraction frequency. In response to the square-wave contraction bout, the increase in Vo(2) to steady state (SS) was 16.7 +/- 1.3 ml x 100 g(-1) x min(-1) (range 13.0-21.9 ml x 100 g(-1) x min(-1); n = 6). The rise in Vo(2) at contractions onset (n = 6) was fit with a time delay (2.1 +/- 1.2 s, range 0.0-7.7 s) plus monoexponential rise to SS (time constant = 9.4 +/- 1.5 s, range 5.2-14.9 s). Furthermore, in two additional myocytes, Vo(2) increased progressively as contraction frequency increased (ramp protocol). This technique for measuring Vo(2) in isolated, single skeletal myocytes represents a novel and powerful investigative tool for gaining mechanistic insight into mitochondrial function and Vo(2) dynamics without potential complications of the circulation and other myocytes.

    View details for DOI 10.1152/japplphysiol.00559.2002

    View details for Web of Science ID 000179815700045

    View details for PubMedID 12391049

  • Preconditioning improves function and recovery of single muscle fibers during severe hypoxia and reoxygenation AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY Kohin, S., Stary, C. M., Howlett, R. A., Hogan, M. C. 2001; 281 (1): C142-C146


    Reperfusion following prolonged ischemia induces cellular damage in whole skeletal muscle models. Ischemic preconditioning attenuates the deleterious effects. We tested whether individual skeletal muscle fibers would be similarly affected by severe hypoxia and reoxygenation (H/R) in the absence of extracellular factors and whether cellular damage could be alleviated by hypoxic preconditioning. Force and free cytosolic Ca2+ ([Ca2+]c) were monitored in Xenopus single muscle fibers (n = 24) contracting tetanically at 0.2 Hz during 5 min of severe hypoxia and 5 min of reoxygenation. Twelve cells were preconditioned by a shorter bout of H/R 1 h before the experimental trial. In preconditioned cells, force relative to initial maximal values (P/P(o)) and relative peak [Ca2+]c fell (P < 0.05) during 5 min of hypoxia and recovered during reoxygenation. In contrast, P/P(o) and relative peak [Ca2+]c fell more during hypoxia (P < 0.05) and recovered less during reoxygenation (P < 0.05) in control cells. The ratio of force to [Ca2+]c was significantly higher in the preconditioned cells during severe hypoxia, suggesting that changes in [Ca2+]c were not solely responsible for the loss in force. We conclude that 1) isolated skeletal muscle fibers contracting in the absence of extracellular factors are susceptible to H/R injury associated with changes in Ca2+ handling; and 2) hypoxic preconditioning improves contractility, Ca2+ handling, and cell recovery during subsequent hypoxic insult.

    View details for Web of Science ID 000169384100017

    View details for PubMedID 11401836

  • Recovery of force during postcontractile depression in single Xenopus muscle fibers AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY Howlett, R. A., Stary, C. M., Hogan, M. C. 2001; 280 (5): R1469-R1475


    This study examined the relationship between force and cytosolic free calcium concentration ([Ca2+]c) in different fiber types from Xenopus before, during, and after cells underwent postcontractile depression (PCD). During a standardized fatigue run, force in the two fast fatiguing (FF) fiber types (types 1 and 2, n = 10) fell more quickly (5.8 vs. 8.1 min) and to a greater degree [0.36 vs. 0.51 of initial (P(o))] than in the slow fatiguing (SF) fiber type (type 3, n = 11). After the initial fatigue run, both FF and SF experienced a drop in force to <15% P(o) (PCD) at a similar time (20.6 vs. 21.4 min). A second stimulation period, undertaken during PCD, produced significant recovery of force in both groups, but significantly more so in SF than FF (64 +/- 7 vs. 29 +/- 2% P(o)). This force recovery during PCD was accompanied by a significant increase in peak [Ca2+]c, particularly in SF. However, despite the significant recovery of force during stimulation while in PCD, the amount of force produced for a given peak [Ca2+]c was significantly lower in both groups during PCD than at any other point in the experiment. A final stimulation period, initiated when all fibers had recovered from PCD, demonstrated a recovery of both force and peak [Ca2+]c in both groups, but this recovery was significantly greater in SF vs. FF. These data demonstrate that with continuous electrical stimulation, it is possible to produce a significant recovery of force production during the normally quiescent period of PCD, but that it occurs with a decreased muscle force production for a given peak [Ca2+]c. This suggests that factors other than structural alterations of the sarcoplasmic reticulum are likely the cause of PCD in these fibers.

    View details for Web of Science ID 000167973400029

    View details for PubMedID 11294770

  • Impairment of Ca2+ release in single Xenopus muscle fibers fatigued at varied extracellular Po-2 JOURNAL OF APPLIED PHYSIOLOGY Stary, C. M., Hogan, M. C. 2000; 88 (5): 1743-1748


    We tested the hypothesis that the mechanisms involved in the more rapid onset of fatigue when O(2) availability is reduced in contracting skeletal muscle are similar to those when O(2) availability is more sufficient. Two series of experiments were performed in isolated, single skeletal muscle fibers from Xenopus laevis. First, relative force and free cytosolic Ca(2+) concentrations ([Ca(2+)](c)) were measured simultaneously in single fibers (n = 6) stimulated at increasing frequencies (0.25, 0.33, 0.5, and 1 Hz) at an extracellular PO(2) of either 22 or 159 Torr. Muscle fatigue (force = 50% of initial peak tension) occurred significantly sooner (P < 0.05) during the low- (237 +/- 40 s) vs. high-PO(2) treatments (280 +/- 38 s). Relative [Ca(2+)](c) was significantly decreased from maximal values at the fatigue time point during both the high- (72 +/- 4%) and low-PO(2) conditions (78 +/- 4%), but no significant difference was observed between the treatments. In the second series of experiments, using the same stimulation regime as the first, fibers (n = 6) exposed to 5 mM caffeine immediately after fatigue demonstrated an immediate but incomplete relative force recovery during both the low- (89 +/- 4%) and high-PO(2) treatments (82 +/- 3%), with no significant difference between treatments. Additionally, there was no significant difference in relative [Ca(2+)](c) between the high- (100 +/- 12% of prefatigue values) and low-PO(2) treatments (108 +/- 12%) on application of caffeine. These results suggest that in isolated, single skeletal muscle fibers, the earlier onset of fatigue that occurred during the low-extracellular PO(2) condition was modulated through similar pathways as the fatigue process during the high and involved a decrease in relative peak [Ca(2+)](c).

    View details for Web of Science ID 000086985000034

    View details for PubMedID 10797138

  • Phosphorylating pathways and fatigue development in contracting Xenopus single skeletal muscle fibers AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY Stary, C. M., Hogan, M. C. 2000; 278 (3): R587-R591


    To investigate the differential contribution of oxidative and substrate-level phosphorylation to force production during repetitive, maximal tetanic contractions, single skeletal muscle fiber performance was examined under conditions of high-oxygen availability and anoxia. Tetanic force development (P) was measured in isolated, single type-1 muscle fibers (fast twitch; n = 6) dissected from Xenopus lumbrical muscle while being stimulated at increasing frequencies (0.25, 0.33, and 0.5 Hz), with each frequency lasting 2 min. Two separate work bouts were conducted, with the perfusate PO(2) being either 0 or 159 mmHg. No significant (P < 0. 05) difference was found in the initial peak tensions (P(0)) between the high (334 +/- 57 kPa) and the low (325 +/- 41 kPa) PO(2) treatment. No significant difference in P was observed between the treatments during the first 50 s. However, a significant difference in force production was observed between the high (P/P(0) = 0.96 +/- 0.02) and the low PO(2) condition (P/P(0) = 0.92 +/- 0.02) by 60 s of work. After 60 s, steady-state force production was maintained during the high compared with the low PO(2) condition until stimulation frequency was increased, at which point developed tension during the high PO(2) condition began to decline. Time to fatigue (P/P(0) = 0.3) was reached significantly sooner during the low (250 +/- 16 s) than the high PO(2) condition (367 +/- 28 s). These results demonstrate that during the first 50 s of 0.25-Hz contractions, substrate-level phosphorylation has the capacity to maintain force and ATP hydrolysis when oxidative phosphorylation is absent. This period was followed by an oxygen-dependent phase in which force generation was maintained during the high PO(2) condition (but not during the low PO(2) condition) until the onset of a final fatiguing phase, at which a calculated maximal rate of oxidative phosphorylation was reached.

    View details for Web of Science ID 000085808000006

    View details for PubMedID 10712276

  • Rapid force recovery in contracting skeletal muscle after brief ischemia is dependent on O-2 availability JOURNAL OF APPLIED PHYSIOLOGY Hogan, M. C., Kohin, S., Stary, C. M., Hepple, R. T. 1999; 87 (6): 2225-2229


    We tested the hypothesis that contracting skeletal muscle can rapidly restore force development during reperfusion after brief total ischemia and that this rapid recovery depends on O(2) availability and not an alternate factor related to blood flow. Isolated canine gastrocnemius muscle (n = 5) was stimulated to contract tetanically (isometric contraction elicited by 8 V, 0.2-ms duration, 200-ms trains, at 50-Hz stimulation) every 2 s until steady-state conditions of muscle blood flow (controlled by pump perfusion) and developed force were attained (3 min). While maintaining the same stimulation pattern, muscle blood flow was then reduced to zero (complete ischemia) for 2 min. Normal blood flow was then restored to the contracting muscle; however, two distinct conditions of oxygenation (at the same blood flow) were sequentially imposed: deoxygenated blood (30 s), blood with normal arterial O(2) content (30 s), a return to deoxygenated blood (30 s), and finally a return to normal arterial O(2) content (90 s). During the ischemic period, force development fell to 39 +/- 6 (SE)% of normal (from 460 +/- 40 to 170 +/- 20 N/100 g). When muscle blood flow was restored to normal by perfusion with deoxygenated blood, developed force continued to decline to 140 +/- 20 N/100 g. Muscle force rapidly recovered to 310 +/- 30 N/100 g (P < 0.05) during the 30 s in which the contracting muscle was perfused with oxygenated blood and then fell again to 180 +/- 30 N/100 g when perfused with blood with low PO(2). These findings demonstrate that contracting skeletal muscle has the capacity for rapid recovery of force development during reperfusion after a short period of complete ischemia and that this recovery depends on O(2) availability and not an alternate factor related to blood flow restoration.

    View details for Web of Science ID 000084182000031

    View details for PubMedID 10601171

  • Pulmonary gas exchange during exercise in pigs JOURNAL OF APPLIED PHYSIOLOGY Hopkins, S. R., Stary, C. M., Falor, E., Wagner, H., Wagner, P. D., McKirnan, M. D. 1999; 86 (1): 93-100


    Increased ventilation-perfusion (VA/Q) inequality is observed in approximately 50% of humans during heavy exercise and contributes to the widening of the alveolar-arterial O2 difference (A-aDO2). Despite extensive investigation, the cause remains unknown. As a first step to more direct examination of this problem, we developed an animal model. Eight Yucatan miniswine were studied at rest and during treadmill exercise at approximately 30, 50, and 85% of maximal O2 consumption (VO2 max). Multiple inert-gas, blood-gas, and metabolic data were obtained. The A-aDO2 increased from 0 +/- 3 (SE) Torr at rest to 14 +/- 2 Torr during the heaviest exercise level, but arterial PO2 (PaO2) remained at resting levels during exercise. There was normal VA/Q inequality [log SD of the perfusion distribution (log) = 0.42 +/- 0.04] at rest, and moderate increases (log = 0.68 +/- 0.04, P < 0.0001) were observed with exercise. This result was reproducible on a separate day. The VA/Q inequality changes are similar to those reported in highly trained humans. However, in swine, unlike in humans, there was no inert gas evidence for pulmonary end-capillary diffusion limitation during heavy exercise; there was no systematic difference in the measured PaO2 and the PaO2 as predicted from the inert gases. These data suggest that the pig animal model is well suited for studying the mechanism of exercise-induced VA/Q inequality.

    View details for Web of Science ID 000080009600014

    View details for PubMedID 9887118

  • Faster adjustment of O-2 delivery does not affect Vo(2) on-kinetics in isolated in situ canine muscle JOURNAL OF APPLIED PHYSIOLOGY Grassi, B., Gladden, L. B., Samaja, M., Stary, C. M., Hogan, M. C. 1998; 85 (4): 1394-1403


    The mechanism(s) limiting muscle O2 uptake (VO2) kinetics was investigated in isolated canine gastrocnemius muscles (n = 7) during transitions from rest to 3 min of electrically stimulated isometric tetanic contractions (200-ms trains, 50 Hz; 1 contraction/2 s; 60-70% of peak V(O2)). Two conditions were mainly compared: 1) spontaneous adjustment of blood flow (Q) [control, spontaneous Q (C Spont)]; and 2) pump-perfused Q, adjusted approximately 15 s before contractions at a constant level corresponding to the steady-state value during contractions in C Spont [faster adjustment of O2 delivery (Fast O2 Delivery)]. During Fast O2 Delivery, 1-2 ml/min of 10(-2) M adenosine were infused intra-arterially to prevent inordinate pressure increases with the elevated Q. The purpose of the study was to determine whether a faster adjustment of O2 delivery would affect V(O2) kinetics. Q was measured continuously; arterial (Ca(O2)) and popliteal venous (Cv(O2)) O2 contents were determined at rest and at 5- to 7-s intervals during contractions; O2 delivery was calculated as Q x Ca(O2), and V(O2) was calculated as Q x arteriovenous O2 content difference. Times to reach 63% of the difference between baseline and steady-state VO2 during contractions were 23.8 +/- 2.0 (SE) s in C Spont and 21.8 +/- 0.9 s in Fast O2 Delivery (not significant). In the present experimental model, elimination of any delay in O2 delivery during the rest-to-contraction transition did not affect muscle V(O2) kinetics, which suggests that this kinetics was mainly set by an intrinsic inertia of oxidative metabolism.

    View details for Web of Science ID 000076306700029

    View details for PubMedID 9760333

  • Peripheral O-2 diffusion does not affect Vo(2) on-kinetics in isolated in situ canine muscle JOURNAL OF APPLIED PHYSIOLOGY Grassi, B., Gladden, L. B., Stary, C. M., Wagner, P. D., Hogan, M. C. 1998; 85 (4): 1404-1412


    To test the hypothesis that muscle O2 uptake (V(O2)) on-kinetics is limited, at least in part, by peripheral O2 diffusion, we determined the V(O2) on-kinetics in 1) normoxia (Control); 2) hyperoxic gas breathing (Hyperoxia); and 3) hyperoxia and the administration of a drug (RSR-13, Allos Therapeutics), which right-shifts the Hb-O2 dissociation curve (Hyperoxia+RSR-13). The study was conducted in isolated canine gastrocnemius muscles (n = 5) during transitions from rest to 3 min of electrically stimulated isometric tetanic contractions (200-ms trains, 50 Hz; 1 contraction/2 s; 60-70% peak V(O2)). In all conditions, before and during contractions, muscle was pump perfused with constantly elevated blood flow (Q), at a level measured at steady state during contractions in preliminary trials with spontaneous Q x Adenosine was infused intra-arterially to prevent inordinate pressure increases with the elevated Q x Q was measured continuously, arterial and popliteal venous O2 concentrations were determined at rest and at 5- to 7-s intervals during contractions, and V(O2) was calculated as Q x arteriovenous O2 content difference. PO2 at 50% HbO2 saturation (P50) was calculated. Mean capillary PO2 (Pc(O2)) was estimated by numerical integration. P50 was higher in Hyperoxia+RSR-13 [40 +/- 1 (SE) Torr] than in Control and in Hyperoxia (31 +/- 1 Torr). After 15 s of contractions, Pc(O2) was higher in Hyperoxia (97 +/- 9 Torr) vs. Control (53 +/- 3 Torr) and in Hyperoxia+RSR-13 (197 +/- 39 Torr) vs. Hyperoxia. The time to reach 63% of the difference between baseline and steady-state V(O2) during contractions was 24.7 +/- 2.7 s in Control, 26.3 +/- 0.8 s in Hyperoxia, and 24.7 +/- 1.1 s in Hyperoxia+RSR-13 (not significant). Enhancement of peripheral O2 diffusion (obtained by increased PcO2 at constant O2 delivery) during the rest-to-contraction (60-70% of peak V(O2)) transition did not affect muscle V(O2) on- kinetics.

    View details for Web of Science ID 000076306700030

    View details for PubMedID 9760334

  • Bioenergetics of contracting skeletal muscle after partial reduction of blood flow JOURNAL OF APPLIED PHYSIOLOGY Hogan, M. C., Gladden, L. B., Grassi, B., Stary, C. M., Samaja, M. 1998; 84 (6): 1882-1888


    The purpose of this study was to examine the bioenergetics and regulation of O2 uptake (VO2) and force production in contracting muscle when blood flow was moderately reduced during a steady-state contractile period. Canine gastrocnemius muscle (n = 5) was isolated, and 3-min stimulation periods of isometric, tetanic contractions were elicited sequentially at rates of 0.25, 0.33, and 0.5 contractions/s (Hz) immediately followed by a reduction of blood flow [ischemic (I) condition] to 46 +/- 3% of the value obtained at 0.5 Hz with normal blood flow. The VO2 of the contracting muscle was significantly (P < 0.05) reduced during the I condition [6.5 +/- 0.8 (SE) ml . 100 g-1 . min-1] compared with the same stimulation frequency with normal flow (11.2 +/- 1.5 ml . 100 g-1 . min-1), as was the tension-time index (79 +/- 12 vs. 123 +/- 22 N . g-1 . min-1, respectively). The ratio of VO2 to tension-time index remained constant throughout all contraction periods. Muscle phosphocreatine concentration, ATP concentration, and lactate efflux were not significantly different during the I condition compared with the 0. 5-Hz condition with normal blood flow. However, at comparable rates of VO2 and tension-time index, muscle phosphocreatine concentration and ATP concentration were significantly less during the I condition compared with normal-flow conditions. These results demonstrate that, in this highly oxidative muscle, the normal balance of O2 supply to force output was maintained during moderate ischemia by downregulation of force production. In addition, 1) the minimal disruption in intracellular homeostasis after the initiation of ischemia was likely a result of steady-state metabolic conditions having already been activated, and 2) the difference in intracellular conditions at comparable rates of VO2 and tension-time index between the normal flow and I condition may have been due to altered intracellular O2 tension.

    View details for Web of Science ID 000073960000009

    View details for PubMedID 9609780

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