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2.
Neuroreport ; 12(7): 1437-41, 2001 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-11388425

RESUMO

We show by immunocytochemistry in frog retina that most members of the Kir subfamily are expressed in specific neuronal compartments. However, Kir 6.1, the pore-forming subunit of K(ATP) channels, is expressed exclusively in glial Müller cells. Müller cell endfeet display strong Kir 6.1 immunolabel throughout the retina, whereas the somata are labeled only in the retinal periphery. This spatial pattern is similar to that of Kir 4.1, of the ratio of inward to outward K+ currents, and of spermine/spermidine immunoreactivity. We suggest that the co-expression of Kir 4.1 and Kir 6.1 subunits may enable the cells to maintain their high K+ conductance and hyperpolarized membrane potentials both at high ATP levels (Kir 4.1) and during ATP deficiency (Kir 6.1).


Assuntos
Potenciais da Membrana/fisiologia , Neuroglia/metabolismo , Canais de Potássio Corretores do Fluxo de Internalização , Canais de Potássio/metabolismo , Rana pipiens/metabolismo , Retina/metabolismo , Visão Ocular/fisiologia , Animais , Especificidade de Anticorpos , Canais de Potássio Corretores do Fluxo de Internalização Acoplados a Proteínas G , Imuno-Histoquímica , Neuroglia/citologia , Potássio/metabolismo , Rana pipiens/anatomia & histologia , Retina/citologia
3.
Brain Res ; 843(1-2): 145-60, 1999 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-10528121

RESUMO

Outwardly rectifying K(+) channels determine the membrane conductance and influence the proliferation rate of glial progenitor cells. To analyze the molecular identity and the functional role of K(+) channels in glial progenitors of mouse brain, expression of shaker-type Kv1 genes was studied at three levels: (1) presence of Kv1 mRNAs, (2) biosynthesis of channel proteins and (3) electrophysiological and pharmacological properties of K(+) currents. mRNA expression of Kv1.1 to Kv1.6 genes was studied by single-cell reverse transcription-mediated polymerase chain reaction (RT-PCR) using degenerate primers to amplify the six Kv1 transcripts. Most cells expressed several mRNA combinations simultaneously. In more than half of the cells, messages for Kv1.2, Kv1.5 and Kv1.6 were found, while Kv1.1, Kv1.3 and Kv1.4 were detected in only a minority of cells. In contrast, at the level of protein expression - employing immunocytochemistry with subtype-specific antibodies - Kv1. 2 and Kv1.3 were undetectable (<2%), while almost all cells expressed Kv1.4 (85%), Kv1.5 (99%) and Kv1.6 (99%). Kv1.1 was present in a minor cell population (10%). Functional contribution of Kv1 proteins to progenitor membrane conductance was determined by analyzing the voltage-dependence of K(+) current activation and inactivation as well as their current sensitivities to the subtype-preferring blockers and toxins tetraethylammonium (TEA), 4-aminopyridine (4-AP), charybdotoxin (CTX), alpha-dendrotoxin (DTX) and mast-cell degranulating peptide (MCDP). From these results, it is concluded: first, glial progenitor cells can express all transcripts of the six Kv1 genes, but do not express all proteins; second, Kv1.4, Kv1.5 and Kv1.6 proteins are most abundant and were found in the majority of cells; and third, K(+) currents flow predominantly either through heteromeric channel complexes or through homomeric Kv1.5 ion pores, but not through homomeric Kv1.4 or Kv1.6 channels.


Assuntos
Neuroglia/fisiologia , Oligodendroglia/fisiologia , Canais de Potássio de Abertura Dependente da Tensão da Membrana , Canais de Potássio/genética , Células-Tronco/fisiologia , Animais , Células Cultivadas , Charibdotoxina/farmacologia , Canais de Potássio de Retificação Tardia , Venenos Elapídicos/farmacologia , Embrião de Mamíferos , Canal de Potássio Kv1.1 , Canal de Potássio Kv1.2 , Canal de Potássio Kv1.3 , Canal de Potássio Kv1.4 , Canal de Potássio Kv1.5 , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos , Neuroglia/citologia , Oligodendroglia/citologia , Fenotiazinas/farmacologia , Canais de Potássio/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células-Tronco/citologia , Tetraetilamônio/farmacologia , Transcrição Gênica
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