Dissociated histaminergic neuron cultures from the tuberomammillary nucleus of rats: culture methods and ghrelin effects
Dissociated histaminergic neuron cultures from the tuberomammillary nucleus of rats: culture methods and ghrelin effects
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DOI：
10.1016/j.jneumeth.2003.09.008
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发表时间：
2004-01-30
影响因子：
3
通讯作者：
Nakajima, Y
中科院分区：
文献类型：
--
作者：
Bajic, D;Hoang, QV;Nakajima, Y
下丘脑的结节乳头核（TMN）是大脑中组胺的唯一来源。这个核通过支配大脑的各个区域，在诸如觉醒和食欲等重要功能中起着重要作用。我们从出生后（3日龄和10日龄）大鼠的颞叶神经中分离出原代组胺能神经元培养物。腺苷脱氨酶（adenosine deaminase， AD）和组胺免疫细胞化学结果显示，我们培养的颞叶神经网络中超过50%的神经元具有组胺能。在大神经元（直径，bbb22）中，组胺能神经元超过88%。这些大神经元（平均直径26.5 μ m）用于电生理。利用大约2个月大的TMN培养物，我们研究了胃饥饿素的作用，胃饥饿素是一种最近发现的刺激食欲的内源性肽。在装载gtpgammas的神经元中，生长素（3mum）抑制了先前由抑制性神经肽nociceptin激活的电流。胃饥饿素平均抑制电流为471 +/- 128 pA （s.e.m., n = 7）。I-V关系揭示了ghrelin抑制的电流是向内整流的，在E-K附近有一个反转电位。这些结果表明ghrelin抑制TMN神经元的G蛋白偶联内向整流K+通道（Kir3， GIRK），并且我们的TMN培养有助于研究脑组胺能神经元的生理特性。(C) 2003 Elsevier B.V.版权所有
The tuberomammillary nucleus (TMN) in the hypothalamus is the sole source of histamine in the brain. This nucleus, by innervating various brain regions, plays an important role for vital functions such as arousal and appetite. We have developed dissociated primary histaminergic neuron cultures from TMN of postnatal (3 and 10-day-old) rats. More than 50% of our cultured neurons from the TMN were histaminergic as revealed by adenosine deaminase (AD) as well as histamine immunocytochemistry. Among large neurons (diameter, >22 mum), more than 88% were histaminergic. Such large neurons (mean diameter, 26.5 mum) were used for electrophysiology. Using about 2-month-old TMN cultures, we investigated the effects of ghrelin, a recently discovered appetite-stimulating endogenous peptide. In GTPgammaS-loaded neurons, ghrelin (3 muM) suppressed currents that had previously been activated by an inhibitory neuropeptide, nociceptin. The mean current suppression by ghrelin was 471 +/- 128 pA (S.E.M., n = 7). The I-V relationship revealed that the ghrelin-suppressed current was inwardly rectifying with a reversal potential around E-K. These results suggest that ghrelin inhibits G protein-coupled inward rectifier K+ channels (Kir3, GIRK) of TMN neurons and that our TMN cultures are useful for investigating physiological properties of brain histaminergic neurons. (C) 2003 Elsevier B.V. All rights reserved.