Carnosine uptake in rat choroid plexus primary cell cultures and choroid plexus whole tissue from PEPT2 null mice.
PEPT2 is functionally active and localized to the apical membrane of rat choroid plexus epithelial cells. However, little is known about the transport mechanisms of endogenous neuropeptides in choroid plexus, and the role of PEPT2 in this process. In the present study, we examined the uptake kinetics of carnosine in rat choroid plexus primary cell cultures and choroid plexus whole tissue from wild-type (PEPT2(+/+)) and null (PEPT2(-/-)) mice. Our results indicate that carnosine is preferentially taken up from the apical as opposed to basolateral membrane of cell monolayers, and that basolateral efflux in limited. Transepithelial flux of carnosine was not distinguishable from that of paracellular diffusion. The apical uptake of carnosine was characterized by a high affinity (K(m) = 34 microM), low capacity (V(max) = 73 pmol/mg protein/min) process, consistent with that of PEPT2. The non-saturable component was small (K(d) = 0.063 microL/mg protein/min) and, under linear conditions, was only 3% of the total uptake. Studies in transgenic mice clearly demonstrated that PEPT2 was responsible for over 90% of carnosine's uptake in choroid plexus whole tissue. These findings elucidate the unique role of PEPT2 in regulating neuropeptide homeostasis at the blood-cerebrospinal fluid interface.
Teuscher
Nathan S
NS
Department of Pharmaceutical Sciences, College of Pharmacy, and Upjohn Center for Clinical Pharmacology, The University of Michigan, Ann Arbor, Michigan 48109, USA.
Shen
Hong
H
Shu
Cathaleen
C
Xiang
Jianming
J
Keep
Richard F
RF
Smith
David E
DE
D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002336:Carnosine / Q000378:metabolism / Q000493:pharmacokinetics*; D002478:Cells, Cultured; D002831:Choroid Plexus / Q000166:cytology / Q000378:metabolism*; D015194:Diffusion Chambers, Culture; D004151:Dipeptides / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D051379:Mice; D018345:Mice, Knockout; D051381:Rats; D027981:Symporters / Q000172:deficiency / Q000235:genetics*
D000596:Amino Acids; D004151:Dipeptides; D027981:Symporters; C093694:hydrogen-coupled oligopeptide transporter PepT2; D002336:Carnosine