MOLECULAR BIOLOGY OF EGG LAYING, A STEREOTYPED BEHAVIOR

这项建议描述了旨在理解控制 神经细胞中的基因表达。以前孤立的和具有特征的 腹型海兔神经肽基因在神经元亚群中的表达 Ganglion将被用作模型系统。体外重组体将是 细菌结构基因B-半乳糖苷酶(B-GAL)之间的构建 或氯霉素乙酰转移酶(CAT)和神经肽基因 推动者包括独联体控制要素。到那时，杂交基因将 海马神经细胞内微量注射BE及其受体表达的研究 评估的B-半乳糖酶或过氧化氢酶。各种各样的神经元，正常情况下 或不表达内源基因将被调查，以 确定显微注射表达的细胞特异性 基因。包括缺失和替换在内的体外突变将是 构建并重新评估了表达的特异性，从而定义了CIS 调节基因表达的作用元件。我们将进一步 显微注射神经肽基因在不同组织中的表达 生理状态。电活动和兴奋性疾病的影响 抑制性神经递质将被研究。体外诱变将 再次用于定义负责响应的独联体作用DNA元件 以改变生理状态。潜在的细胞内信使 将对调解这些反应的因素进行调查。在这些候选人中 信使有cAMP、钙等离子。 上述研究将确定分子遗传机制。 在发育过程中被大脑利用，还将有助于产生 在分子遗传水平上对神经可塑性的理解。这 研究将进一步加深我们对各种神经元疾病的理解 从儿童的学习障碍到阿尔茨海默病 老年人。

This proposal describes experiments aimed at understanding the control of gene expression in nerve cells. Previously isolated and characterized neuropeptide genes expressed in subsets of neurons in the Aplysia abdominal ganglion will be used as a model system. In vitro recombinants will be constructed between the bacterial structural genes B-galactosidase (B-gal) or chloramphenicol acetyl transferase (CAT) and the neuropeptide gene promoters including CIS controlling elements. The hybrid genes will then be microinjected into Aplysia neurons in organ culture and expression of the B-gal or CAT enzymes assessed. A variety of neurons which normally do or do not express the endogenous gene will be investigated in order to ascertain the cellular specificity of the expression of microinjected genes. In vitro mutations including deletions and substitutions will be constructed and the specificity of expression reassessed thus defining CIS acting elements regulating expression of the genes. We will further investigate expression of the microinjected neuropeptide genes in different physiological states. The effects of electrical activity and excitatory or inhibitory neurotransmitters will be studied. In vitro mutagenesis will again be used to define CIS acting DNA elements responsible for responding to alternate physiological conditions. Potential intracellular messengers which mediate the responses will be investigated. Among these candidate messengers are cAMP, calcium and other ions. The studies described above will define molecular genetic mechanisms utilized by the brain during development and will also help generate an understanding of neuroplasticity at the molecular genetic level. This research will further our understanding of diverse neuronal disorders ranging from learning disabilities in children to Alzheimer's disease in the aged.