STRESS ALLOSTASIS: CRF, SEROTONIN AND THE BNST

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在2007年，我们完成了我们的研究表明，多种5 HT受体表达在不同的亚群BNST神经元。我们不仅有生理数据显示单个BNST神经元可以表达一种以上的受体，在2007年，我们还开发了一种技术（单细胞RT-PCR），从而我们可以从单个神经元中提取mRNA并检查其遗传5 HT受体谱。 我们还完成了对单个BNST神经元表达的内在膜电流的检查。 我们在BNST中鉴定了至少三种不同的细胞类型（I - III型）。在电流钳模式下，这三种亚型显示响应于去极化电流注入的特征性动作电位放电模式，以及显示对超极化电流注入的不同电压响应。 电流钳数据表明，三种亚型的BNST神经元可以进一步区分基于其表达模式的特定的内在膜电流。 我们还开始研究形成BNST神经元放电模式的离子通道的遗传基础。 我们采用相同的单细胞RT-PCR技术来检测BNST神经元中表达的离子通道亚基。 到目前为止，我们已经确定了至少5个IA通道亚基，3个IT通道亚基和2个IH通道亚基。 这些亚基中的每一个具有不同的动力学特征和传导性，以及在细胞内的差异分布。 因此，这些不同的离子通道亚基的组合可以微妙地，但显着地影响这些神经元的放电模式。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In 2007 we completed our study demonstrating that multiple 5HT receptors are expressed in distinct subpopulations of BNST neurons. Not only do we have physiologic data showing that individual BNST neurons can express more than one receptor, in 2007 we also developed a technique (single cell RT-PCR) whereby we can extract the mRNA from individual neurons and examine their genetic 5HT receptor profile. We also completed our examination of the intrinsic membrane currents that are expressed by individual BNST neurons. We identified at least three distinct cell types in the BNST (Types I - III). In current-clamp mode, these three subtypes show characteristic action potential firing patterns in response to depolarizing current injection, as well as showing a distinct voltage response to hyperpolarizing current injection. The current clamp data suggested that the three subtypes of BNST neuron could be further differentiated based on their expression patterns for specific intrinsic membrane currents. We also began examination of the genetic underpinning for the ion channels that shape the firing pattern of BNST neurons. We employed the same single cell RT-PCR technique to examine the ion channel subunits that are expressed in BNST neurons. To date we have characterized at least 5 IA channel subunits, 3 IT channel subunits and 2 IH channel subunits. Each of these subunits has a different kinetic profile and conductance, as well as a differential distribution within the cell. Hence, the combination of these different ion channel subunits can subtly, but dramatically, influence the firing pattern of these neurons.