Spontaneous and GABA-induced single channel currents in cultured murine spinal cord neurons

Abstract

Intracellular and patch clamp recordings were made from embryonic mouse spinal cord neurons growing in primary cell culture. Outside-out membrane patches obtained from these cells usually showed spontaneous single channel currents when studied at the resting potential (−56 ± 1.5 mV). In 18 out of 30 patches tested, spontaneous single channel activity was abolished by making Tris⁺ the major cation on both sides of the membrane. The remaining patches continued to display spontaneous single channel currents under these conditions. These events reversed polarity at a patch potential of 0 mV and displayed a mean single channel conductance of 24 ± 1.2 pS. Application of the putative inhibitory transmitter γ-aminobutyric acid (0.5–10 μM) to outside-out patches of spinal cord cell membrane induced single channel currents in 10 out of 15 patches tested. These channels had a primary conductance of 29 ± 2.8 pS in symmetrical 145 mM Cl solutions. Frequency distributions for the open times of these channels were well fit by the sum of a fast exponential term ("of") with a time constant τ_of = 4 ± 1.3 ms and a slow exponential term ("os") with a time constant τ_os = 24 ± 8.1 ms. Frequency distributions for channel closed times were also well fit by a double exponential equation, with time constants τ_cf = 2 ± 0.2 ms and τ_cs = 62 ± 20.9 ms.