MOLECULAR REGULATION OF NEURONAL GAP JUNCTIONS DURING DEVELOPMENT AND INJURY

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。 在哺乳动物神经系统中，神经元通过间隙连接（GJ;电突触）的耦合在胚胎和/或出生后早期发育期间增加，并且在许多发育事件中起重要作用，包括神经元分化、细胞死亡、突触发生和神经回路形成。GJ耦合随后减少，并保持在成人低，仅限于特定的神经元子集。在成熟的神经系统中，GJ耦合在许多病理条件下增加，包括创伤性和缺血性损伤。然而，在发育过程中神经元GJ耦合增加的机制， 受伤情况不明。将在大鼠下丘脑神经元培养物的拟议研究中对其进行研究。首先，我们将检验这样一个假设，即在发育过程中，神经元GJ偶联的增加是由GABA-A受体和II组代谢型谷氨酸受体通过CREB依赖性调节的。 调节主要的神经元连接蛋白，连接蛋白36（Cx 36）。这将使用分子生物学、染料偶联、蛋白质印迹、北方印迹和活神经元染色进行检测。第二、 我们将检验这样的假设，即负责神经元间隙连接的发育上调的机制也负责神经元损伤期间间隙连接的上调。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。这项研究可能有助于了解神经元损伤的机制，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.