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Am J Physiol Cell Physiol 267: C563-C569, 1994;
0363-6143/94 $5.00
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AJP - Cell Physiology, Vol 267, Issue 2 C563-C569, Copyright © 1994 by American Physiological Society

ARTICLES

Intercellular communication between mouse Leydig cells

W. A. Varanda and A. C. de Carvalho
Department of Physiology, Faculty of Medicine of Ribeirao Preto, Universidade de Sao Paulo, Brazil.

In this paper we describe the basic features of gap junctions in pairs of Leydig cells mechanically dissociated from mouse testes, studied with the double whole cell patch-clamp technique. These cells are extensively coupled with regard to dye injection and electrophysiological measurements. The mean junctional conductance (gj) measured in 61 pairs of cells was 10.6 +/- 1.5 (SE) nS. In most pairs gj was voltage dependent when transjunctional voltage exceeded +/- 50 mV. On imposition of a voltage gradient across the junction the transjunctional current decayed exponentially to a lower level, with a time constant of 3.3 s at 50 mV and 430 ms at 100 mV. As in other systems, octanol (600 microM final concentration) uncoupled the cells within approximately 2 min. In a few cell pairs, gj was low enough to permit recording of single channel currents without the use of uncoupling agents. Single channel conductance fluctuations measured using pipettes containing potassium aspartate were distributed mainly around three peaks, at 21, 39, and 60 pS, suggesting the presence of channels formed by connexin 43. Western blots of Percoll gradient purified Leydig cells using specific antibodies indicate that connexin 43 is indeed expressed in these cells, whereas connexin 26 and connexin 32 are not.


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