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Immortalized Gonadotropin-Releasing Hormone Neurons (GT1-7 Cells) Exhibit Synchronous Bursts of Action Potentials

Toshiya Funabashi a Kumiko Suyama a Tsuguo Uemura b Makiko Hirose b Fumiki Hirahara b Fukuko Kimura a
Neuroendocrinology , (2001); 73 :157-165

Departments of a) Physiology, and b) Gynecology and Obstetrics, Yokohama City University School of Medicine,Yokohama, Japan

Although it has been assumed that synchronized firing of gonadotropin-releasing hormone (GnRH) neurons is necessary for pulsatile GnRH secretion, there is no clear evidence for this. In the present study we simultaneously recorded spontaneous action potentials from multiple cells. Immortalized GnRH neurons (GT1-7 cells) were cultured on a multi-electrode dish (MED) and action potentials recorded by an extracellular recording method. One to two weeks after the beginning of culture, spontaneous action potentials appeared, exhibiting bursts composed of 5-10 action potentials. Burst activity was intermittent and periodic with mean burst intervals of 13.3 s. Further-more, burst activity was recorded almost simultaneously from several micro-electrodes, suggesting that electrical activities of GT1-7 cells were synchronized with each other. Periodic bursts were completely and reversibly blocked by 1-5 Ì M tetrodotoxin, indicating that voltage-dependent Na + channels are involved in their generation. Á -Aminobutyric acid (GABA) given at a 10-Ì M concentra-tion shortened inter-burst intervals, whereas 10 Ì M bicuculline lengthened them. Finally, the gap junctional blockers n-octyl alcohol (1 mM) and carbenoxolone (100 Ì M) reversibly blocked periodic burst activity. The present study provides direct evidence that the electrical activity of GT1-7 cells exhibits synchronous and periodic bursts composed of action potentials. In addition, endogenous GABA is involved in GT1-7 cells in determining burst frequency. Although the precise mechanism of synchronized burst activities needs to be clarified, gap junctional communications among GT1-7 cells are at least partially involved.

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