title: Expression of nucleoside transporter in freshly isolated neurons and astrocytes from mouse brain.
abstract: Nucleoside transporters comprise equilibrative ENT1-4 and concentrative CNT1-3. CNTs transport against an intracellular/extracellular gradient and are essential for transmitter removal, independently of metabolic need. ENT1-4 mediate transport until intracellular/extracellular equilibrium of the transported compound, but are very efficient, when the accumulated nucleoside or nucleobase is rapidly eliminated by metabolism. Most nucleoside transporters are membrane-bound, but ENT3 is mainly intracellular. This study uses freshly isolated neurons and astrocytes from two adult mouse strains. In one transgenic strain the neuronal marker Thy1 was associated with a compound fluorescing at one wavelength, and in the other the astrocytic marker GFAP was associated with a compound fluorescent at a different wavelength. Highly purified astrocytic and neuronal populations (as determined by presence/absence of cell-specific genes) were obtained from these mice by fluorescence-activated cell sorting. In each population mRNA analysis was performed by reverse-transcription polymerase chain reaction. CNT1 was absent in both cell types; all other nucleoside transporters were expressed to at least a similar degree (in relation to applied amount of RNA and to a house-keeping gene) in astrocytes as in neurons. Astrocytic ENT3 enrichment was dramatic, but it was not up-regulated after fluoxetine-mediated increase in DNA synthesis. A comparison with results obtained in cultured astrocytes shows that the latter are generally compatible with the present findings and suggests that many observations obtained in intact tissue, mainly by in situ hybridization (which also determines mRNA expression) may underestimate astrocytic nucleoside transporter expression.
authors.affiliation: Department of Clinical Pharmacology and Institute of Pathology and Pathophysiology, China Medical University, No. 92 Beier Road, Heping District, Shenyang, People's Republic of China.
authors.forename: B
authors.initials: B
authors.lastname: Li
authors.forename: L
authors.initials: L
authors.lastname: Gu
authors.forename: L
authors.initials: L
authors.lastname: Hertz
authors.forename: L
authors.initials: L
authors.lastname: Peng
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D033721:Equilibrative Nucleoside Transporter 1 / Q000096:biosynthesis; D005260:Female; D005473:Fluoxetine / Q000494:pharmacology; D008297:Male; D026901:Membrane Transport Proteins / Q000096:biosynthesis; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D033703:Nucleoside Transport Proteins / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D033721:Equilibrative Nucleoside Transporter 1; D005473:Fluoxetine; D026901:Membrane Transport Proteins; D033703:Nucleoside Transport Proteins; D012333:RNA, Messenger; C501425:Slc29a3 protein, mouse; C498476:SLC29A1 protein, mouse; C065171:cif nucleoside transporter
references.citation: Cereb Cortex. 2012 May;22(5):1203-14
references.pmid: 21810784
references.citation: Neuropharmacology. 2009 Feb;56(2):429-37
references.pmid: 18957298
references.citation: J Neurosci. 2008 Jan 2;28(1):264-78
references.pmid: 18171944
references.citation: Acta Otolaryngol. 2008 Aug;128(8):936-40
references.pmid: 18607998
references.citation: Cereb Cortex. 2014 Jan;24(1):67-80
references.pmid: 22997174
references.citation: Neuroscience. 2006 Sep 29;142(1):125-37
references.pmid: 16859834
references.citation: Proc Natl Acad Sci U S A. 2012 Apr 17;109(16):6265-70
references.pmid: 22421436
references.citation: J Biol Chem. 2005 Apr 22;280(16):15880-7
references.pmid: 15701636
references.citation: Neurochem Int. 2012 Dec;61(8):1325-32
references.pmid: 23017600
references.citation: Fundam Clin Pharmacol. 2014 Apr;28(2):217-20
references.pmid: 23130601
references.citation: PLoS Genet. 2010 Feb 05;6(2):e1000833
references.pmid: 20140240
references.citation: Neurosci Lett. 1996 Aug 9;213(3):153-6
references.pmid: 8873137
references.citation: J Neurosci. 2004 Jan 21;24(3):692-701
references.pmid: 14736855
references.citation: Pharmacol Ther. 2007 Dec;116(3):401-16
references.pmid: 17884172
references.citation: J Neurosci. 2000 Dec 15;20(24):9104-10
references.pmid: 11124987
references.citation: Biochem Biophys Res Commun. 2013 Aug 16;438(1):145-51
references.pmid: 23876310
references.citation: Neurochem Res. 2012 Nov;37(11):2480-95
references.pmid: 22711334
references.citation: Neuroscience. 2007 May 25;146(3):1193-211
references.pmid: 17408864
references.citation: J Neurosci. 1998 Nov 1;18(21):8794-804
references.pmid: 9786986
references.citation: J Neurosci Res. 2006 Sep;84(4):801-8
references.pmid: 16862552
references.citation: Brain Res Bull. 1987 Oct;19(4):473-84
references.pmid: 3690362
references.citation: Leukemia. 2000 Dec;14(12):2118-27
references.pmid: 11187901
references.citation: Psychopharmacology (Berl). 2009 Nov;207(1):1-12
references.pmid: 19662385
references.citation: J Physiol. 2007 Jul 1;582(Pt 1):137-51
references.pmid: 17446223
references.citation: J Biol Chem. 2001 Jan 26;276(4):2914-27
references.pmid: 11032837
references.citation: Neuropharmacology. 2009 Sep;57(4):375-85
references.pmid: 19596362
references.citation: J Neurosci Res. 1989 Oct;24(2):260-7
references.pmid: 2585550
references.citation: Front Biosci (Elite Ed). 2011 Jun 01;3:896-900
references.pmid: 21622101
references.citation: Glia. 2005 Oct;52(1):25-35
references.pmid: 15892125
references.citation: Ann N Y Acad Sci. 2009 Jan;1152:30-42
references.pmid: 19161374
references.citation: J Biol Chem. 2004 Nov 26;279(48):50042-9
references.pmid: 15448143
references.citation: Prog Neuropsychopharmacol Biol Psychiatry. 1984;8(4-6):487-93
references.pmid: 6152337
references.citation: Biochem Biophys Res Commun. 2005 Sep 9;334(4):1343-50
references.pmid: 16043124
references.citation: Psychopharmacology (Berl). 2008 Dec;201(3):443-58
references.pmid: 18758753
references.citation: Curr Top Med Chem. 2011;11(8):948-72
references.pmid: 21401500
references.citation: Neurochem Res. 2002 Feb;27(1-2):113-20
references.pmid: 11930908
references.citation: Eur Arch Psychiatry Clin Neurosci. 2007 Aug;257(5):300-3
references.pmid: 17401725
references.citation: Methods Mol Biol. 2012;814:305-23
references.pmid: 22144315
references.citation: J Neurosci. 2007 Nov 7;27(45):12255-66
references.pmid: 17989291
references.citation: Methods. 1998 Nov;16(3):293-310
references.pmid: 10071068
references.citation: Epilepsia. 2011 Sep;52(9):1645-55
references.pmid: 21635241
references.citation: Drug Metab Dispos. 2004 Dec;32(12):1455-61
references.pmid: 15371301
references.citation: Neurochem Res. 2012 Nov;37(11):2351-63
references.pmid: 22528833
references.citation: Neurochem Res. 2010 Sep;35(9):1434-44
references.pmid: 20577800
references.citation: Nat Cell Biol. 2007 Aug;9(8):945-53
references.pmid: 17618272
references.citation: Neurochem Res. 2012 Nov;37(11):2569-88
references.pmid: 22926576