MOLECULAR REGULATION OF NEURONAL GAP JUNCTIONS DURING DEVELOPMENT AND INJURY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在哺乳动物神经系统中，神经元通过缝隙连接(GJS；电突触)连接 在胚胎和/或出生后早期发育期间增加，并在 发育事件的数量，包括神经元分化、细胞死亡、突触形成和 神经回路的形成。在受限的成人中，GJ偶联随后降低并保持在低水平 特定的神经元亚群。在成熟的神经系统中，GJ偶联在 多种病理情况，包括创伤性和缺血性损伤。然而， 在发育和发育过程中神经元GJ偶联增加的机制 受伤情况尚不清楚。它们将在拟议的大鼠下丘脑神经元研究中进行研究。 文化。首先，我们将检验在发育过程中神经元GJ偶联增加的假设 受GABA-A受体和第二组代谢性谷氨酸受体通过CREB依赖调节 主要神经元连接蛋白Cx36的调节。这将使用以下工具进行测试 分子生物学、染料偶联、Western印迹、Northern印迹和活神经元染色。第二, 我们将检验这样一种假设，即负责发育上调的机制 神经元缝隙连接也是神经元损伤过程中缝隙连接上调的原因。 因此，GABA-A受体、第二类代谢性谷氨酸受体和CREB在 将测试神经元损伤介导的缝隙连接偶联增加。低渗性休克 将其作为体外神经元损伤模型。拟议的研究可能有助于理解 研究神经元损伤的机制，并开发治疗损伤的新方法。

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.