A monocarboxylate transporter MCT1 is located at the basolateral pole of rat jejunum.
We have functionally expressed and identified a monocarboxylate transporter (MCT1) from rat jejunal enterocyte and we provide evidence for its basolateral localization. Poly(A)+ RNA isolated from rat jejunum was injected into Xenopus laevis oocytes and expression of a proton-lactate symporter was investigated by means of L-[14C]lactate uptake. The existence of an endogenous capacity for L-lactate transport was demonstrated; when, however, oocytes were injected with jejunal mRNA, an expressed L-lactate uptake was seen which differed from the endogenous transporter since it was significantly pH dependent. After sucrose density gradient fractionation, the highest expression of the pH-dependent lactate uptake was detected with the mRNA size fraction of about 2-3 kb in length. The substrate specificity, stereoselectivity and sensitivity to pCMBS (an organomercurial thiol reagent that modifies cysteine residues) of the expressed transport were in good agreement with results previously obtained using isolated jejunal basolateral membranes. Using the reverse transcriptase-polymerase chain reaction, the presence of mRNA coding for the MCT1 isoform was demonstrated in jejunal enterocytes. These data, together with previous results, suggest that MCT1 is a major route for lactate efflux across the basolateral membrane of rat jejunum; this is in contrast to current opinion which restricts the presence of MCT1 to the apical membrane of the whole small intestine.
Orsenigo
M N
MN
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, via Celoria 26, I-20133 Milano, Italy. novella.orsenigo@unimi.it
Tosco
M
M
Bazzini
C
C
Laforenza
U
U
Faelli
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007583:Jejunum / Q000378:metabolism*; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D008845:Microinjections; D008969:Molecular Sequence Data; D027501:Monocarboxylic Acid Transporters; D009865:Oocytes / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014982:Xenopus laevis; D020245:p-Chloromercuribenzoic Acid / Q000494:pharmacology
D002352:Carrier Proteins; D027501:Monocarboxylic Acid Transporters; D012333:RNA, Messenger; D019344:Lactic Acid; D020245:p-Chloromercuribenzoic Acid