Functional Astrocyte Leak Potassium Channels

星形胶质细胞是一种星形细胞，长期以来被认为在功能上为中枢神经系统（CNS）中的神经元提供结构、内务和代谢支持。这一假设是基于星形胶质细胞不能像神经元那样通过电脉冲传递信号这一事实。然而，星形胶质细胞确实表现出通过特征不佳的钾通道携带的膜电流。虽然这些水流在40多年前就被发现了，但这些通道的性质在很大程度上仍然未知。该提案的重点是确定星形胶质细胞电流是否通过新发现的钾通道家族进行，称为双孔结构域钾通道（2PKs）。本项目将进一步探讨星形胶质细胞2PKs的变化如何调节星形胶质细胞的稳态功能，进而影响神经元功能。星形胶质细胞的膜离子通道电流将在急性大鼠海马切片中测量。潜在的星形胶质细胞2PKs将在基因水平上通过单细胞RT-PCR进一步探索，在蛋白水平上通过共聚焦免疫细胞化学特异性抗体标记后星形胶质细胞。星形细胞2PKs的功能影响将通过在器官型培养海马中使用RNAi沉默鉴定的2PKs来确定。这些实验将描述这些通道的作用，并阐明星形胶质细胞的一般功能。由于星形胶质细胞在神经系统对损伤或疾病的反应中起着不可或缺的作用，因此这些知识对人类健康以及对神经系统的基本理解具有重要意义。该项目将为本科生、研究生和博士后提供参与星形胶质细胞研究的机会。更广泛的影响被认为是强大的，特别是考虑到PI所在的学院没有现成的本科生通道。

Astrocytes are star-shaped cells that have long been functionally considered to provide structural, housekeeping and metabolic support to neurons in the central nervous system (CNS). This supposition has been based on the fact that astrocytes are not able to relay signals by means of electric impulses, as do neurons. However, astrocytes do exhibit membrane currents that are carried through poorly characterized potassium channels. Although these currents were discovered over 40 years ago, the nature of these channels remains largely unknown. This proposal is focused on determining whether astrocyte currents are carried through a a newly discovery potassium channel family, called two-pore domain potassium channels (2PKs). This project will further explore how changes in astrocyte 2PKs could modulate astrocyte homeostatic functions with consequent effect on neuronal function. Membrane ion channel currents from astrocytes will be measured in acute rat hippocampal slices measure. Potential astrocyte 2PKs will be further explored at the gene level by using single cell RT-PCR, and at the protein level by specific antibody labeling to post astrocytes using confocal immunocytochemistry. The functional impact of astrocytic 2PKs will be determined by silencing identified 2PKs using RNAi in the organotypic cultured hippocampus. These experiments will delineate the role of these channels and clarify the function of astrocytes in general. Since astrocytes play integral roles response of the nervous system to injury or disease, such knowledge has implications for human health as well as for the fundamental understanding of the nervous system. The PI will provide opportunities for undergraduate, graduate, and postdoctoral students to participate in research on astrocytes. The broader impacts were judged as strong, particularly considering the PI is in an institute without ready access to undergraduate students.