WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Neurokinin B activates synchronized intracellular Ca2+ oscillations in KNDy neurons obtained from the hypothalamic arcuate nucleus of Kiss1–GFP transgenic mice
Kana Ikegami 1 , Shiori Minabe 1 , Nahoko Ieda 1 , Teppei Goto 1 , Hitomi Abe 1 , Makoto Sanbo 2 , Masumi Hirabayashi 2 , Kei-ichiro Maeda 3 , Hiroko Tsukamura 1 & Yoshihisa Uenoyama 1
1Nagoya University, Nagoya, Japan; 2National Institute for Physiological Science, Okazaki, Japan; 3The University of Tokyo, Bunkyo-ku, Japan.
Pulsatile secretion of GnRH/LH is indispensable for puberty onset and normal reproductive functions in mammalian species. A cohort of neurons expressing three neuropeptides, kisspeptin, neurokinin B (NKB), and dynorphin (KNDy neurons), localized in the hypothalamic arcuate nucleus (ARC), are considered to be a source of GnRH pulse generator. A synchronous discharge of KNDy neurons might be obligatory for pulsatile GnRH secretion. The present study aimed to determine whether NKB–NK3R signaling is required for synchronized activities in KNDy neurons.KNDy–GFP cells taken from the ARC of the Kiss1–GFP transgenic mice embryos at days 17–18 of embryonic age were cultured on a glass-base dish for 3–6 weeks. Intracellular Ca2+ concentrations were measured in individual KNDy–GFP cells, which were superfused with/without 1 μM senktide, a NKB receptor agonist, using the fluorescent Ca2+ indicator Fura-PE3. Frequent and synchronized intracellular Ca2+ oscillations were observed in cultured KNDy–GFP cells with senktide, whereas few Ca2+ oscillations were found without senktide. The senktide-induced Ca2+ oscillations were synchronized in the neighboring KNDy–GFP cells. Such Ca2+oscillations were abolished by chelating extracellular Ca2+ with EGTA, suggesting that Ca2+ oscillations are caused by an influx of extracellular Ca2+ through the calcium channels in KNDy–GFP cells. These results indicate that NKB–NK3R signaling facilitates synchronized activities in neighboring KNDy neurons. This work was supported in part by the Research Program on Innovative Technologies for Animal Breeding, Reproduction, and Vaccine Development.
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ISSN 2052-1472 (online)
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