Lactic acidosis in vivo: Testing the link between lactate generation and H+ accumulation in ischemic mouse muscle
Department of Radiology, Box 357115, University of Washington Medical Center, Seattle, WA 98195-7115, USA. Journal of Applied Physiology
(Impact Factor: 3.06).
06/2010; 108(6):1479-86. DOI: 10.1152/japplphysiol.01189.2009
The link between lactate generation and cellular acidosis has been questioned based on the possibility of H+ generation, independent of lactate production during glycolysis under physiological conditions. Here we test whether glycolytic H+ generation matches lactate production over a physiological pH and lactate range using ischemia applied to the hindlimb of a mouse. We measured the H+ generation and ATP level in vivo using 31P-magnetic resonance spectroscopy and chemically determined intracellular lactate level in the hindlimb muscles. No significant change was found in ATP content by chemical analysis (P>0.1), in agreement with the stoichiometric decline in phosphocreatine (20.2+/-1.2 mM) vs. rise in Pi (18.7+/-2.0 mM), as measured by 31P-magnetic resonance spectroscopy. A substantial drop in pH from 7.0 to 6.7 and lactate accumulation to 25 mM were found during 25 min of ischemia. The rise in H+ generation closely agreed with the accumulation of lactate, as shown by a close correlation with a slope near identity (0.98; r2=0.86). This agreement between glycolytic H+ production and elevation of lactate is confirmed by an analysis of the underlying reactions involved in glycolysis in vivo and supports the concept of lactic acidosis under conditions that substantially elevate lactate and drop pH. However, this link is expected to fail with conditions that deplete phosphocreatine, leading to net ATP hydrolysis and nonglycolytic H+ generation. Thus both direct measurements and an analysis of the stoichiometry of glycolysis in vivo support lactate acidosis as a robust concept for physiological conditions of the muscle cell.
Available from: Alan R Prescott
- "Ischaemia results in a pH decline in muscle tissue  and the translocation to the Z- and I-bands of resident sHSPs [96,97], which is the location of the desmin intermediate filaments. Therefore, the data we have presented evidence the importance of pH changes to the interaction of CRYAB with desmin. "
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ABSTRACT: CRYAB (αB-crystallin) is expressed in many tissues and yet the R120G mutation in CRYAB causes tissue-specific pathologies, namely cardiomyopathy and cataract. Here, we present evidence to demonstrate that there is a specific functional interaction of CRYAB with desmin intermediate filaments that predisposes myocytes to disease caused by the R120G mutation. We use a variety of biochemical and biophysical techniques to show that plant, animal and ascidian small heat-shock proteins (sHSPs) can interact with intermediate filaments. Nevertheless, the mutation R120G in CRYAB does specifically change that interaction when compared with equivalent substitutions in HSP27 (R140G) and into the Caenorhabditis elegans HSP16.2 (R95G). By transient transfection, we show that R120G CRYAB specifically promotes intermediate filament aggregation in MCF7 cells. The transient transfection of R120G CRYAB alone has no significant effect upon cell viability, although bundling of the endogenous intermediate filament network occurs and the mitochondria are concentrated into the perinuclear region. The combination of R120G CRYAB co-transfected with wild-type desmin, however, causes a significant reduction in cell viability. Therefore, we suggest that while there is an innate ability of sHSPs to interact with and to bind to intermediate filaments, it is the specific combination of desmin and CRYAB that compromises cell viability and this is potentially the key to the muscle pathology caused by the R120G CRYAB.
Available from: Norbert Maassen
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ABSTRACT: During and after two successive incremental cycle ergometer tests (tests A and B), plasma potassium concentration ([K+]p), plasma pH (pHp), plasma partial pressure of carbon dioxide, blood lactate concentration ([Lac-]b) and ventilation (VE) were measured. While there was a good correlation between the increase in [K+]p and VE or pHp, respectively, in test A, in test B a close correlation was found only between the increase in VE and [K+]p (r greater than 0.9 for nearly all single cases; r was 0.84 and 0.89 for all (pooled) cases in tests A and B, respectively; the correlation coefficients between changes in pHp and VE in tests A and B were r = 0.74 and r = 0.28, respectively, and r = 0.89 and r = 0.10 between the changes in [Lac-]b and VE in tests A and B). The close relationship for individuals between VE and [K+]p in tests A and B supported the hypothesis that the extracellular increase in [K+] may contribute to the ventilatory drive during exercise. The comparison of the results of tests A and B further indicated that the relationship between pHp and VE was dependent on the experimental design, and that pHp and VE changes are unlikely to be cause and effect.
Available from: George A Brooks
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