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Titolo:
CLONING, EXPRESSION, AND LOCALIZATION OF A MOUSE RETINAL GAMMA-AMINOBUTYRIC-ACID TRANSPORTER
Autore:
RUIZ M; EGAL H; SARTHY V; QIAN XJ; SARKAR HK;
Indirizzi:
BAYLOR COLL MED,DEPT MOLEC PHYSIOL & BIOPHYS,1 BAYLOR PLAZA HOUSTON TX 77030 BAYLOR COLL MED,DEPT MOLEC PHYSIOL & BIOPHYS HOUSTON TX 77030 NORTHWESTERN UNIV,SCH MED,DEPT OPHTHALMOL CHICAGO IL 60611
Titolo Testata:
Investigative ophthalmology & visual science
fascicolo: 12, volume: 35, anno: 1994,
pagine: 4039 - 4048
SICI:
0146-0404(1994)35:12<4039:CEALOA>2.0.ZU;2-8
Fonte:
ISI
Lingua:
ENG
Soggetto:
BRAIN GABA TRANSPORTER; CAT RETINA; GLUTAMIC-ACID; MESSENGER-RNA; RAT RETINA; NEUROTRANSMITTER TRANSPORTERS; FUNCTIONAL EXPRESSION; RABBIT RETINA; CELLS; DNA;
Keywords:
GABA TRANSPORTER; CLONING AND EXPRESSION; IN SITU LOCALIZATION; RETINAL NEUROTRANSMITTER;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
54
Recensione:
Indirizzi per estratti:
Citazione:
M. Ruiz et al., "CLONING, EXPRESSION, AND LOCALIZATION OF A MOUSE RETINAL GAMMA-AMINOBUTYRIC-ACID TRANSPORTER", Investigative ophthalmology & visual science, 35(12), 1994, pp. 4039-4048

Abstract

Purpose. To isolate a cDNA clone encoding a high-affinity gamma-aminobutyric acid (GABA) transporter from mouse retina, to examine its biochemical and pharmacologic properties, and to determine the sites of its mRNA expression in retinal cells. Methods. A mouse retinal cDNA library was screened using a fragment of a rat brain GABA transporter (GAT-1) cDNA as a probe. One homologous clone, mouse retinal GAT-1, was chosen for further characterization. RNA transcribed from mouse retinal GAT-1 was microinjected into Xaopus oocytes, and pharmacologic properties of the expressed transporter were determined. Sites of mouse retinal GAT-1 mRNA expression were examined by in situ hybridization. Results. The protein sequence deduced from the DNA sequence of mouse retinal GAT-1 cDNA was virtually identical to that of the rat and the mouse brain GAT-1. RNA transcribed from this clone induced a [H-3]-GABA uptake activity in microinjected Xenopus oocytes that was both sodium and chloride dependent. The apparent K, and V,, for the GABA uptake were 8.3 mu M and 40.0 pmol/egg per hour, respectively. The mouse retinal GAT-I induced GABA uptake was inhibited by L-diaminobutyric acid, guvacine, cis-4-hydroxynipecotic acid, nipecotic acid, and 4,5,6,7-tetrahydroisoxazolo [4,5c]-pyridin-3-ol with IC50 values of 320, 79, 71, 7.1, and 200 mu M, respectively. However, beta-alanine was unable to inhibitthe induced GABA uptake significantly (IC50 approximate to 2,500 mu M). In situ hybridization studies showed that mouse retinal GAT-1 mRNA was present in a subpopulation of amacrine, interplexiform, and displaced amacrine cells. Hybridization signal in the Muller cells was significantly lower, and GAT-1 transcripts were not detected in the bipolar, horizontal, or photoreceptor cells of mouse retina. Conclusions. Themouse retinal GAT-I cDNA encodes a Na+-dependent, high-affinity GABA transporter that is mainly expressed in a subset of mouse retinal inter neurons.

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Documento generato il 01/04/20 alle ore 20:56:45