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Volume 16, Number 10, Issue of May 15, 1996 pp. 3287-3295
Copyright ©1996 Society for Neuroscience

Temporal Regulation of Shaker- and Shab-Like Potassium Channel Gene Expression in Single Embryonic Spinal Neurons during K⁺ Current Development

Received Oct. 6, 1995; revised Feb. 26, 1996; accepted Feb. 28, 1996.

Devorah Gurantz¹, Angeles B. Ribera², and Nicholas C. Spitzer¹

¹ Department of Biology and Center for Molecular Genetics, University of California San Diego, La Jolla, California 92093-0357, and ² Department of Physiology, University of Colorado Health Sciences Center, Denver, Colorado 80262

A developmental increase in density of delayed rectifier potassium current (I_Kv) in embryonic Xenopus spinal neurons shortens action potential durations and limits calcium influx governing neuronal differentiation. Although previous work demonstrates that maturation of I_Kv depends on general mRNA synthesis, it is not known whether increases in K⁺ channel gene transcripts direct maturation of the current. Accordingly, the developmental appearance of specific Kv potassium channel genes was determined using single-cell reverse transcription-PCR techniques after whole-cell recording of I_Kv during the period of its development. Detection of a coexpressed housekeeping gene along with the potassium channel gene controlled for successful aspiration of cellular mRNA and allowed scoring of cells in which Kv gene transcripts were not detected. Diverse types of Xenopus spinal neurons exhibit homogenous development of I_Kv both in vivo and in culture. In contrast, transcripts of two genes encoding delayed rectifier current, Kv1.1 (Shaker) and Kv2.2 (Shab), are expressed heterogeneously during the period in which the current develops. Kv1.1 mRNA achieves maximal appearance in ~30% of cells, while I_Kv is immature; Kv2.2 mRNA appears later in ~60% of mature neurons. Kv1.1 and 2.2 are thus candidates for generation of I_Kv, and spinal neurons are a heterogeneous population with respect to potassium channel gene expression. Moreover, correlation of gene expression with current properties shows that neurons lacking Kv2.2 have a characteristic voltage dependence of activation of I_Kv.

Key words: potassium current; RT-PCR; single cell gene expression; Kv genes; spinal neurons; Xenopus embryo

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