RESUMO
Previous studies in retinal glial (Müller) cells have suggested that (1) the dominant membrane currents are mediated by K(+) inward-rectifier (Kir) channels (Newman and Reichenbach, Trends Neurosci 19:307-312, 1996), and (2) rectification of these Kir channels is due largely to a block of outward currents by endogenous polyamines such as spermine/spermidine (SPM/SPD) (Lopatin et al., Nature 372:366-369, 1994). In frog Müller cells, the degree of rectification of Kir-mediated currents is significantly higher in the endfoot than in the somatic membrane (Skatchkov et al., Glia 27:171-181, 1999). This article shows that in these cells there is a topographical correlation between the local cytoplasmic SPM/SPD immunoreactivity and the ratio of inward to outward K(+) currents through the surrounding membrane area. Throughout the retina, Müller cell endfeet display a high SPM/SPD immunolabel (assessed by densitometry) and a large inward rectification of K(+) currents, as measured by the ratio of inward to outward current produced by step changes in [K(+)](o). In the retinal periphery, Müller cell somata are characterized by roughly one-half of the SPM/SPD immunoreactivity and K(+)-current rectification as the corresponding endfeet. In the retinal center, Müller cell somata are virtually devoid of both SPM/SPD immunolabel and K(+)-current inward rectification. Comparing one region of the retina with another, we find an exponential correlation between the local K(+) rectification and the local SPM/SPD content. This finding suggests that the degree of inward rectification in a given membrane area is determined by the local cytoplasmic polyamine concentration.
Assuntos
Neuroglia/metabolismo , Canais de Potássio/fisiologia , Retina/metabolismo , Espermidina/metabolismo , Espermina/metabolismo , Animais , Condutividade Elétrica , Eletrofisiologia , Imuno-Histoquímica/métodos , Rana pipiens , Retina/citologia , Coloração e Rotulagem , Distribuição TecidualRESUMO
The radial glial cells that span the retina, described by Müller in 1851, have a remarkable distribution of ion channels in adult amphibia that mediate extracellular K+ spatial buffering. 94% of the total membrane conductance of these cells resides in inward rectifier K+ channels in the endfoot processes apposed to the vitreous humour. We now report that this regional specialization is found in Müller cells isolated from adult (>120 day old) bullfrogs but to a far less extent in those from 10-20 day old tadpoles (stages 34-36). Using the cell attached configuration of the patch-clamp technique, we found, in agreement with previous studies in salamanders, that the endfoot of adult cells had 19.2+/-2.4 (mean +/- S.E., n = 81) channels/patch, whereas the soma had 1.81+/-0.28 (n = 21) channels/patch. In the tadpole, the respective values were 4.29+/-0.26 (n = 79) for the endfoot and 2.26+/-0.24 (n = 27) for the soma. The slope conductance of the inward rectifier K+ channel in 115 mM K+, 19.2+/-0.25 pS (n = 205), channel kinetics and the resting membrane potential (-69+/-2.7 mV, n = 224) were similar at both the endfoot and soma of both adults and embryos. We conclude that during development, the K+ conductance of the Müller cell endfoot, but not of the soma, increases due to a selective clustering of inwardly rectifying K+ channels in that specific region of the cell membrane. The properties of the channels change little during the transformation from tadpole to adult bullfrog.
Assuntos
Neuroglia/metabolismo , Canais de Potássio/metabolismo , Retina/citologia , Retina/metabolismo , Animais , Técnicas In Vitro , Cinética , Larva/fisiologia , Potenciais da Membrana/fisiologia , Condução Nervosa/fisiologia , Neuroglia/ultraestrutura , Técnicas de Patch-Clamp , Rana catesbeiana , Retina/crescimento & desenvolvimentoRESUMO
Macroscopic voltage-dependent currents were recorded from the surface of the intact optic nerve of Rana pipiens using the loose patch clamp technique. Depolarizing steps of more than 40 mV produced sodium-dependent TTX sensitive inward currents and a 4-AP and sodium sensitive fast outward current in addition to a slower outward current. Since the surface of the nerve is a glia limitans, it appears that the membranes of these astrocytes contain both voltage-sensitive sodium and potassium channels.