MOLECULAR DESCRIPTION OF THE CORTEX AND HIPPOCAMPUS

我们发展了消减杂交技术作为一种 用于分离受限制的mRNA克隆的强大工具 解剖学上定义的神经元子集。使用这种技术，我们有 分离出rds神经元变性基因的产物。我们建议 测试模型作为腔内粘连功能的研究 分子，以提高我们对rds处置的理解。 蛋白质在外段盘膜上，证明失活 的rds蛋白是rds表型的唯一决定因素， 充分了解rds诱变事件和突变体的性质 蛋白质表达。我们还分离了编码a基因的mRNAs克隆。 端脑富含钙调蛋白结合底物的蛋白激酶 C和一种端脑富含钙离子的钙调蛋白 激活剂。为了解释每一个的生理意义，我们将 绘制它们的基因图谱，以了解它们是否是 已知的基因，突变体已经被表征，我们还将 产生受其表达影响的刻意突变的小鼠。对这件事 最后，我们将开发核酶和同源重组基因 淘汰法。我们将生产重组蛋白用于 对功能的生化评估，包括证明 真的有这种活性并寻找它的天然底物。我们 将研究将蛋白质定向到树突的氨基酸序列。

We have developed the technique of subtractive hybridization as a powerful tool for isolating clones of mRNAs that are restricted to anatomically-defined subsets of neurons. Using this technique we have isolated the product of the rds neuronal degeneration gene. We propose studies to test a model for its function as an intralumenal adhesion molecule, to improve our understanding of the disposition of the rds protein in the outer segment disc membrane, to prove that inactivation of the rds protein is solely responsible for the rds phenotype, and to understand fully the nature of the rds mutagenic event and mutant protein expression. We have also isolated clones of mRNAs encoding a telencephalon-enriched, calmodulin-binding substrate for protein kinase C and a telencephalon-enriched Ca++, calmodulin-dependent protein kinase. To interpret the physiological significance of each, we will map their genes so as to learn whether they are the products of any known gene for which mutants have been characterized and we will also produce deliberate mouse mutants affected in their expression. To this end, we will develop ribozyme and homologous recombination gene knock-out methodologies. We will produce recombinant proteins for biochemical assessments of function, including proof that the kinase truly has that activity and a search for its natural substrates. We will investigate amino acid sequences that direct proteins to dendrites.