Relations between the stage of cell maturation and lactate transporter activities in rat neonatal muscle cells in culture.
Lactate transport was investigated in newborn rat muscle cells in culture. The aim was to study the lactate transport function at two stages of cell differentiation in culture: (i) during the proliferative phase characterized by myoblasts and myotubes (MyB/MyT2) obtained after 2-3 seedings, (ii) when myotubes (MyT1) grow old in culture after 8-9 seedings. In both developmental stages MyB/MyT2, lactate was carried following a saturable and sigmoidal velocity curve: the Hill and the Scatchard plot analyses confirmed an allosteric or multisite mechanism of lactate transport with two classes of carriers: one of low and one of high affinity i.e., 8.6 and 0.95 mm, respectively, which are associated with high and low transport capacities (V(m)) i.e., 9.1 and 0.67 nm/min/mg, respectively. With MyT1, the velocity curve of lactate transport presented a hyperbolic profile, and the Hill plot analysis gave a Hill number near one suggesting that for cell aging in culture the decrease in cooperativity shows that lactate transport essentially occurs through the low affinity transport system. Inhibitor effects also contributed to evidence for at least two systems of transport. Results obtained from primary cells give evidence for the early activity of lactate transport system at the Myb/MyT2 stage and its evolution during cell aging in culture (MyT1). Sarcolemmal lactate transport in primary cultures of myocytes is accomplished by multiple carriers, neither of which are MCT1 or MCT2 as confirmed by immunoblots.
Beaudry
M
M
Laboratoire de Physiologie des Adaptations UFR MEDECINE COCHIN. Université René Descartes, 24, Rue du faubourg Saint-Jacques 75014-Paris, France.
Mouaffak
N
N
Darribere
T
T
El Abida
K
K
Rieu
M
M
Mengual
R
R
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000494:Allosteric Regulation; D000818:Animals; D000831:Animals, Newborn; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D016922:Cellular Senescence; D003373:Coumaric Acids / Q000494:pharmacology; D015151:Immunoblotting; D027501:Monocarboxylic Acid Transporters; D024510:Muscle Development; D018482:Muscle, Skeletal / Q000254:growth & development* / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
D002352:Carrier Proteins; D003373:Coumaric Acids; D027501:Monocarboxylic Acid Transporters; D013439:Sulfhydryl Reagents; C007175:alpha-cyano-4-hydroxycinnamate; D020245:p-Chloromercuribenzoic Acid; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid