ROLE OF GAP-43 IN OLFACTORY DEVELOPMENT AND LEARNING

GAP-43 在嗅觉发育和学习中的作用

• 批准号: 6108509
• 负责人: RACHAEL Lee NEVE
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6108509
• 关键词: RNA; animal developmental psychology; antisense nucleic acid; behavioral /social science research tag; developmental neurobiology; dopamine; early experience; gene expression; genetically modified animals; immunocytochemistry; in situ hybridization; isozymes; laboratory mouse; laboratory rat; learning; mutant; neuron component; neurotransmitters; northern blottings; olfactory lobe; olfactory stimulus; phosphoproteins; preference; protein kinase C; transfection

The neuronal growth associated protein GAP-43 has been implicated in the modulation of neural plasticity during development and learning. We propose to examine and manipulate the expression of GAP-43 in the olfactory bulb during early learning. First, we will examine the expression of GAP- 43 in the olfactory bulb during postnatal development by RNA blots, RNAse protection quantification, in situ hybridization, and immunocytochemistry. Because phosphorylation of GAP-43 by protein kinase C is critical for the function of this protein, and because recent data suggest that protein kinase C may stabilize GAP-43 mRNA, we will also analyze the expression of the protein kinase C isozymes in the bulb during development. These same analyses will be performed on the olfactory bulbs from animals that have undergone olfactory preference training. Extracellular dopamine has been shown to increase in the neonatal olfactory bulb during early olfactory preference acquisition. Data from our laboratory suggests that the release of dopamine is modulated by GAP-43. Hence, we postulate a link between GAP-43 expression and dopamine levels in the olfactory bulb. Proposed genetic intervention experiments to test for this relationship include injection of Herpes Simplex Virus One (HSV-1) GAP-43 recombinants in vivo into the olfactory bulb and the construction of transgenic mice expressing a GAP-43 mutant. We will use HSV-1 vectors expressing GAP-43 antisense RNA to decrease the expression of GAP-43 in dopaminergic glomerular-layer cells in the olfactory bulb during olfactory preference learning, and assay the injected rats by 2-deoxyglucose uptake, in vivo dialysis, and behavioral preference to determine the consequences of such genetic intervention. We will also use HSV-1 vectors to overexpress GAP-43 in the olfactory bulb after the sensitive period for olfactory preference learning, to determine whether this manipulation will allow expression of any of the neural or behavioral correlates of early odor preference acquisition. Finally, we will construct transgenic mice expressing, under the control of the tyrosine hydroxylase promoter, a mutant of GAP-43 in which the serine-41 that is normally phosphorylated by protein kinase C is replaced by a glycine residue. The aim is to cause deficiency in dopamine release in dopaminergic neurons expressing this mutant. We will examine the effects of such a mutation on early olfactory learning in transgenic mice.

神经元生长相关蛋白GAP-43参与了神经元的生长。 调节发育和学习过程中的神经可塑性。 我们 目的：检测和调控GAP-43在嗅觉神经元中的表达， 在早期的学习过程中。 首先，我们将检查GAP的表达- 43在出生后发育过程中嗅球中的表达 保护定量、原位杂交和免疫细胞化学。 由于蛋白激酶C对GAP-43的磷酸化对于细胞的生长至关重要， 这种蛋白质的功能，因为最近的数据表明，蛋白质 激酶C可以稳定GAP-43 mRNA，我们还将分析 鳞茎发育过程中蛋白激酶C同工酶的变化。 这些相同 将对来自具有以下特征的动物的嗅球进行分析： 进行嗅觉偏好训练。 细胞外多巴胺 显示在新生儿嗅球中的早期嗅球增加 偏好获取 我们实验室的数据表明， 多巴胺是由GAP-43调节的。 因此，我们假设 嗅球中GAP-43表达和多巴胺水平。 提出 测试这种关系的遗传干预实验包括 体内注射单纯疱疹病毒1（HSV-1）GAP-43重组体 导入嗅球并构建表达 GAP-43突变体 我们将使用表达GAP-43反义RNA的HSV-1载体 降低多巴胺能肾小球层细胞GAP-43的表达 在嗅觉偏好学习过程中， 注射大鼠2-脱氧葡萄糖摄取，体内透析，和行为 我们倾向于确定这种基因干预的后果。 我们 还将使用HSV-1载体在嗅球中过表达GAP-43 嗅觉偏好学习敏感期后， 这种操作是否会允许表达任何神经或 早期气味偏好获得的行为相关性。 最后我们 将构建转基因小鼠表达，在控制下， 酪氨酸羟化酶启动子，GAP-43的突变体，其中丝氨酸-41 通常被蛋白激酶C磷酸化的蛋白被一个 甘氨酸残基 目的是造成多巴胺释放不足， 表达这种突变体的多巴胺能神经元。 我们将检验 这种突变对转基因小鼠早期嗅觉学习的影响。