MOLECULAR REGULATION OF NEURONAL GAP JUNCTIONS DURING DEVELOPMENT AND INJURY

该子项目是利用 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。 在哺乳动物神经系统中，神经元通过缝隙连接（GJ;电突触）的耦合。 在胚胎和/或出生后早期发育期间增加，并在 许多发育事件，包括神经元分化、细胞死亡、突触发生和 神经回路的形成GJ耦合随后降低，并在成人中保持较低水平， 到特定的神经元子集。在成熟的神经系统中，GJ耦合在发育过程中增加 许多病理状况，包括创伤性和缺血性损伤。但 在发育过程中负责神经元GJ耦合增加的机制， 受伤情况不明。它们将在大鼠下丘脑神经元的拟议研究中进行研究 cultures.首先，我们将测试在发育过程中神经元GJ耦合增加的假设， 由GABA-A受体和II组代谢型谷氨酸受体通过CREB依赖性调节 调节主要的神经元连接蛋白，连接蛋白36（Cx 36）。这将通过以下方式进行测试： 分子生物学、染料偶联、蛋白质印迹、北方印迹和活神经元染色。第二、 我们将检验这一假设，即负责发育上调的机制 在神经元损伤过程中，神经元缝隙连接的表达也是缝隙连接上调的原因。 因此，GABA-A受体、II组代谢型谷氨酸受体和CREB在 将测试神经元损伤介导的间隙连接偶联的增加。低渗休克 将用作体外神经元损伤模型。拟议的研究可能有助于了解 神经元损伤的机制，并开发新的方法来治疗损伤。

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 the mammalian nervous system, coupling of neurons by gap junctions (GJs; electrical synapses) increases during embryonic and/or early postnatal development and plays an important role in a number of developmental events, including neuronal differentiation, cell death, synaptogenesis and neural circuit formation. GJ coupling subsequently decreases and remains low in the adult, confined to specific subsets of neurons. In the mature nervous system, GJ coupling increases during a number of pathological conditions, including traumatic and ischemic injuries. However, the mechanisms that are responsible for increases in neuronal GJ coupling during development and injuries are unknown. They will be studied in the proposed research in rat hypothalamic neuronal cultures. First, we will test the hypothesis that increase in neuronal GJ coupling during development is regulated by GABA-A receptors and group II metabotropic glutamate receptors via CREBdependent regulation of the main neuronal connexin, connexin 36 (Cx36). This will be tested using molecular biology, dye coupling, western blots, northern blots, and staining for live neurons. Second, we will test the hypothesis that the mechanisms that are responsible for developmental up-regulation of neuronal gap junctions also are responsible for gap junction up-regulation during neuronal injury. Therefore, the role of GABA-A receptors, group II metabotropic glutamate receptors, and CREB in the neuronal injury-mediated increases in gap junction coupling will be tested. Hypoosmotic shock will be used as an in vitro neuronal injury model. The proposed research may help to understand the mechanisms of neuronal injuries and to develop new approaches for the treatment of injuries.