Expression of amino acid transport systems in Xenopus oocytes injected with mRNA of rat small intestine and kidney.
Xenopus and Cynops oocytes were injected with exogenous mRNA prepared from rat small intestine and kidney and their electrical responses to amino acids were measured by both the current clamped and the voltage clamped methods. Oocytes injected with mRNA of rat small intestine showed a depolarization response to several neutral and basic amino acids, and almost no response to acidic amino acids. The responses to amino acids increased with incubation time after injection of mRNA, and followed Michaelis-Menten type kinetics. The responses were dependent on both Na+ concentration and membrane potential, and were inactivated by a sulfhydryl reagent, 5,5-dithiobis(2-nitrobenzoate). These results are interpreted as due to the expression of Na+/amino acid cotransporter(s) in oocytes injected with rat small intestine mRNA. On the other hand, the oocyte injected with rat kidney mRNA showed a hyperpolarization response to neutral amino acids, a depolarization response to basic ones, and almost no response to acidic ones in frog Ringer solution. These responses were independent of Na+ concentration and followed Michaelis-Menten type kinetics. These amino acid response characteristics in oocytes injected with rat kidney mRNA are interpreted as due to the expression of facilitated diffusion carrier protein(s) (uniporter) of amino acids in the oocyte.
Aoshima
H
H
Department of Chemistry, Faculty of Liberal Arts, Yamaguchi University, Japan.
Tomita
K
K
Sugio
S
S
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D002021:Buffers; D005260:Female; D007421:Intestine, Small / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008564:Membrane Potentials; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D012455:Salamandridae; D013439:Sulfhydryl Reagents; D014982:Xenopus laevis
D000596:Amino Acids; D002021:Buffers; D012333:RNA, Messenger; D013439:Sulfhydryl Reagents