CALCINEURIN IN CALCIUM CHANNEL REGULATION DURING AGING.

描述：(申请人描述) 电压敏感的钙通道(VSCCs)的增加促成了这种变化 在衰老过程中的神经元功能，并可能使易感性 神经退行性变。最近的证据表明，细胞内的 蛋白磷酸酶，钙调神经磷酸酶(CaN)，在海马区增加 年事已高。此外，抑制剂是否能降低培养的VSCC电流 海马细胞。因此，拟议的实验将探索这部小说 假设CAN和VSCC的年龄相关变化在功能上是相关的。 在第一组研究中，部分分离的(拉链)切片 结合细胞附着式膜片钳技术的准备将是 用于确定CaN抑制剂是否对海马VSCC有不同的影响 老年和年轻成年大鼠的活动。这一准备工作还允许 提取单个的、具有生理特征的神经元，主要 树突状突起完整，用于后续的RT-PCR分析。通过这种方式，可以 同一细胞中的mRNA水平和VSCC活性可用于确定是否可以 (或CaN调节因子)和VSCCs随年龄的变化方向相同。 最后，将全细胞记录和钙离子成像应用于培养的 海马细胞确定是否可以调节细胞内钙信号 通过调制VSCCS。总之，这些研究应该会增加我们的理解 CaN在钙调节和脑老化中的作用。

DESCRIPTION: (Applicant's Description) An increase in voltage-sensitive Ca2+ channels (VSCCS) contributes to changes in neuronal function during aging, and may render vulnerability to neurodegeneration. Recent evidence indicates that cytosolic levels of the protein phosphatase, calcineurin (CaN), increase in the hippocampus with advanced age. Moreover, CaN inhibitors reduce VSCC currents in cultured hippocampal cells. As such, the proposed experiments will explore the novel hypothesis that age-related changes in CaN and VSCCs are functionally related. In the first set of studies, the partially dissociated ("zipper") slice preparation in combination with the cell-attached patch clamp technique will be used to determine whether CaN inhibitors differentially affect hippocampal VSCC activity in aged and young adult rats. This preparation also permits the extraction of individual, physiologically-characterized neurons, with major dendritic processes intact, for subsequent RT-PCR analysis. In this manner, CaN MRNA levels and VSCC activity in the same cell can be used to determine if CaN (or CaN-regulated factors) and VSCCs change in the same direction with age. Finally, whole-cell recording and Ca2+ imaging will be applied to cultured hippocampal cells to determine if CaN regulates intracellular Ca2+ signalling by modulating VSCCS. Together, these studies should increase our understanding of the role of CaN in Ca2+ regulation and brain aging.