BIOCHEMICAL STUDIES OF NEURONS AND OTHER CELL TYPES

神经元和其他细胞类型的生物化学研究

• 批准号: 6107965
• 负责人: Douglas Eric Brenneman
• 金额: --
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6107965
• 关键词: HIV envelope protein gp120; autoradiography; cell cycle; complementary DNA; cyclins; developmental neurobiology; in situ hybridization; insulinlike growth factor; molecular cloning; neural plate /tube; neurons; neuroprotectants; neurotrophic factors; neutralizing antibody; nucleic acid sequence; tissue /cell culture; vasoactive intestinal peptide

The neurotrophic properties of a nine amino acid core peptide (ADNF-9) derived from activity dependent neurotrophic factor was investigated in cell cultures produced from the rat cerebral cortex. Comparative studies of ADNF-9 action in mixed (glia plus neurons) versus glia-depleted neuronal cultures indicated that ADNF-9 can act directly on neurons, although the potency of the peptide was 10,000-fold greater in mixed cultures. Kinetic studies showed that exposure to ADNF-9 for only two hours was sufficient to produce a four day protection against the cell killing action of tetrodotoxin. Treatment with bafilomycin A1 for two hours prevented this ADNF-9-mediated neuroprotection. Antiserum to ADNF increased the number of apoptotic neurons in cerebral cortical cultures and decreased the activity of choline acetyltransferase in spinal cord cultures. ADNP, a cloned protein with shared epitopes and biological activity with ADNF, was localized in the embryonic and adult CNS on the mouse by in situ hybridization. ADNP mRNA was detected in the trophoblast and the placenta, with intense labeling in the floor plate. In adult brain, ADNP mRNA was especially intense in the olfactory bulb, thalamus, hippocampus and cerebellum. Administration of anti-ADNF to pregnant mice significantly inhibited embryonic growth and produced microcephaly. Progeny from anti-ADNF-treated mothers exhibited significant decreases in performance on an incremental DRL schedule learning paradigm when tested as adults. VIP is a secretagogue of ADNF. In the pregnant mouse, a transient increase in mRNA for VIP was found in the trophoblast at E6 and E7. VIP binding sites were abundant in the E6 trophoblast and in the embryonic neuroepithelium and decidua throughout pregnancy. VIP mRNA was not detected in the mouse embryo until E11-12. Early pregnancy factor (EPF) mRNA was detected throughout the brains of adult mice, but this greater intensity in the thalamic nuclei and cortex during pregnancy. EPF mRNA was also present in the embryonic neuroepithelium and the surrounding amniotic and trophoblastic membranes from E6 to E12. An active peptide from EPF was shown to prevent the growth-inhibitory effects of gp120, the envelope protein from the human immunodeficiency virus.

九个氨基酸核心肽 (ADNF-9) 的神经营养特性 源自活性依赖性神经营养因子的研究 从大鼠大脑皮层产生的细胞培养物。比较研究 ADNF-9 在混合（神经胶质细胞加神经元）与神经胶质细胞耗尽中的作用 神经元培养表明 ADNF-9 可以直接作用于神经元， 尽管混合肽的效力高出 10,000 倍 文化。 动力学研究表明，仅暴露于 ADNF-9 两次 小时足以产生四天的细胞保护作用 河豚毒素的杀伤作用。 巴弗洛霉素 A1 治疗两人 数小时阻止了 ADNF-9 介导的神经保护作用。 ADNF 抗血清 增加大脑皮层培养物中凋亡神经元的数量 并降低脊髓胆碱乙酰转移酶的活性 文化。 ADNP，一种具有共享表位和生物学特性的克隆蛋白 ADNF 的活性，定位于胚胎和成人 CNS 小鼠通过原位杂交。在滋养层中检测到 ADNP mRNA 和胎盘，底板上有强烈的标签。 成人时 在大脑中，ADNP mRNA 在嗅球、丘脑、 海马体和小脑。 给怀孕小鼠施用抗 ADNF 显着抑制胚胎生长并产生小头畸形。 经抗 ADNF 治疗的母亲的后代表现出显着下降 测试时增量 DRL 计划学习范式的性能 作为成年人。 VIP 是 ADNF 的促分泌剂。 在怀孕的老鼠身上， 在 E6 时滋养层中发现 VIP mRNA 短暂增加 和E7。 VIP 结合位点在 E6 滋养层和 整个怀孕期间的胚胎神经上皮和蜕膜。 VIP mRNA 为 直到 E11-12 才在小鼠胚胎中检测到。 早孕因素 (EPF) mRNA 在成年小鼠的大脑中被检测到，但这 怀孕期间丘脑核和皮质的强度更大。 EPF mRNA 也存在于胚胎神经上皮和 从 E6 到 E12 周围的羊膜和滋养层。 一个 EPF 的活性肽被证明可以防止生长抑制 gp120（人类免疫缺陷的包膜蛋白）的影响 病毒。