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
基本信息
- 批准号:6290144
- 负责人:
- 金额:--
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:
- 资助国家:美国
- 起止时间:至
- 项目状态:未结题
- 来源:
- 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
项目摘要
The neurotrophic and growth-stimulating actions of vasoactive intestinal peptide (VIP) are mediated indirectly through secreted, glia-derived substances. Our focus is to characterize these VIP-mediators and study their mechanism of action. A principal glial mediator of VIPs neuroprotective actions is activity dependent neurotrophic factor (ADNF). Previous studies indicated that ADNF exhibited neuroprotection at femtomolar concentrations and had a structural similarity to heat shock protein 60 (hsp60). However, our recent studies indicate that ADNF preparations tested over a very broad range of concentrations exhibited two peaks of survival-promoting activity when tested in cerebral cortical cultures co-treated with tetrodotoxin. The EC50s of these activities differed by 100,000-fold. Although no ADNF heterogeneity was evident with five chromatographic techniques, analysis of purified ADNF by capillary electrophoresis (CE) demonstrated two peaks, each exhibiting survival-promoting activity, only one of which was blocked by antiserum to hsp60. Amino acid sequence data was obtained after trypsin digestion of ADNF. A total of 15.8 kDa was sequenced, with none of the peptides matching any known sequence. Antisera generated against one of the digest peptides produced neuronal cell death when incubated with cerebral cortical cultures and blocked the survival-promoting activity of ADNF and the non-hsp60 peak isolated by CE. These data suggest that ADNF is more complex than previously recognized and may consist of two active components, one sharing an epitope with hsp60, the other being a unique protein/subunit. To further explore the protective actions of VIP-related proteins, a peptide associated with ADNF and another peptide obtained from a related protein (activity dependent neuroprotective protein) were used to prevent embryonic death and growth restriction in a model of fetal alcohol syndrome. Furthermore, the protective actions of the peptides were effective even one hour after the administration of alcohol. VIP has a dramatic effect on mouse embryonic growth at the time of neural tube closure. In further investigations of the mediators of VIP-stimulated growth, isolated neural tube from embryonic day 9.5 mice were incubated with VIP and the conditioned medium (CM) studied. As analyzed by CE, 35 peaks were detected in CM. In this complex mixture, two cytokines were identified: interleukin-1 and interleukin-6. These cytokines have been shown to have proliferative and neurotrophic roles in the developing CNS. - VIP, PACAP, astrocytes, cytokines, stress proteins, neuroprotection, neural tube, fetal alcohol syndrome, trophic factors
血管活性肠肽(VIP)的神经营养和生长刺激作用是通过分泌的胶质源性物质间接介导的。我们的重点是表征这些VIP介质,并研究其作用机制。VIP神经保护作用的主要胶质介质是活性依赖性神经营养因子(ADNF)。先前的研究表明,ADNF在飞摩尔浓度下表现出神经保护作用,并且与热休克蛋白60(hsp 60)具有结构相似性。然而,我们最近的研究表明,ADNF制剂测试在一个非常广泛的浓度范围内表现出两个峰值的生存促进活动时,在大脑皮层文化共同处理河豚毒素。这些活性的EC 50相差100,000倍。虽然没有ADNF的异质性是明显的五个色谱技术,纯化的ADNF的毛细管电泳(CE)分析表明,两个峰,每个表现出生存促进活性,只有其中之一被阻断的hsp 60的抗血清。胰蛋白酶消化ADNF后获得氨基酸序列数据。对总共15.8kDa的肽进行测序,没有肽匹配任何已知序列。产生的抗血清对消化肽之一产生神经元细胞死亡时,与大脑皮层培养物孵育,并阻止ADNF的生存促进活性和非热休克蛋白60峰分离CE。这些数据表明,ADNF比以前认识到的更复杂,可能由两个活性成分组成,一个与HSP 60共享表位,另一个是独特的蛋白质/亚基。为了进一步探索VIP相关蛋白的保护作用,使用与ADNF相关的肽和从相关蛋白(活性依赖性神经保护蛋白)获得的另一种肽来防止胎儿酒精综合征模型中的胚胎死亡和生长受限。此外,肽的保护作用甚至在给予酒精后一小时也是有效的。VIP在神经管闭合时对小鼠胚胎生长具有显著影响。在VIP刺激生长的介质的进一步调查中,从胚胎9.5天的小鼠分离的神经管与VIP和条件培养基(CM)一起孵育。通过CE分析,CM中检测到35个峰。在这种复杂的混合物中,鉴定了两种细胞因子:白细胞介素-1和白细胞介素-6。这些细胞因子已被证明在发育中的CNS中具有增殖和神经营养作用。- VIP、PACAP、星形胶质细胞、细胞因子、应激蛋白、神经保护、神经管、胎儿酒精综合征、营养因子
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Douglas Eric Brenneman其他文献
Douglas Eric Brenneman的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Douglas Eric Brenneman', 18)}}的其他基金
Development of KLS-13019 for Neuropathic Pain
开发用于治疗神经性疼痛的 KLS-13019
- 批准号:
10326595 - 财政年份:2021
- 资助金额:
-- - 项目类别:
Development of KLS-13019 for Neuropathic Pain
开发用于治疗神经性疼痛的 KLS-13019
- 批准号:
10493291 - 财政年份:2021
- 资助金额:
-- - 项目类别:
Development of KLS-13019 for Neuropathic Pain
开发用于治疗神经性疼痛的 KLS-13019
- 批准号:
10704175 - 财政年份:2021
- 资助金额:
-- - 项目类别:
Novel Thioderivatives as Neuroprotective Anticonvulsants
作为神经保护抗惊厥药的新型硫代衍生物
- 批准号:
7745751 - 财政年份:2009
- 资助金额:
-- - 项目类别:
相似海外基金
Development of non-contact autoradiography technology using magnetic field
利用磁场的非接触式放射自显影技术的开发
- 批准号:
19K15947 - 财政年份:2019
- 资助金额:
-- - 项目类别:
Grant-in-Aid for Early-Career Scientists
Ultra High Resolution Brain PET Scanner for in-vivo Autoradiography Imaging
用于体内放射自显影成像的超高分辨率脑 PET 扫描仪
- 批准号:
9791189 - 财政年份:2018
- 资助金额:
-- - 项目类别:
Ultra High Resolution Brain PET Scanner for in-vivo Autoradiography Imaging
用于体内放射自显影成像的超高分辨率脑 PET 扫描仪
- 批准号:
10117728 - 财政年份:2018
- 资助金额:
-- - 项目类别:
Ultra High Resolution Brain PET Scanner for in-vivo Autoradiography Imaging
用于体内放射自显影成像的超高分辨率脑 PET 扫描仪
- 批准号:
10237144 - 财政年份:2018
- 资助金额:
-- - 项目类别:
Ultra High Resolution Brain PET Scanner for in-vivo Autoradiography Imaging
用于体内放射自显影成像的超高分辨率脑 PET 扫描仪
- 批准号:
10436281 - 财政年份:2018
- 资助金额:
-- - 项目类别:
Development of autoradiography technique capable of sequential imaging -Toward visualization of radioactive cesium dynamics in botanical bodies-
开发能够连续成像的放射自显影技术 -实现植物体中放射性铯动力学的可视化 -
- 批准号:
17K05109 - 财政年份:2017
- 资助金额:
-- - 项目类别:
Grant-in-Aid for Scientific Research (C)
application of high resolution arufa-autoradiography for analysing micro-distribution of B-compund in tissues and its BNCT effects
应用高分辨率阿鲁法放射自显影分析B化合物在组织中的微观分布及其BNCT效应
- 批准号:
26670557 - 财政年份:2014
- 资助金额:
-- - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Visualization of the change in pain by the autoradiography and calcium imaging in the brain and spinal cord
通过放射自显影和大脑和脊髓钙成像显示疼痛变化
- 批准号:
24659294 - 财政年份:2012
- 资助金额:
-- - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Digital Autoradiography of an Irradiated Fuel Pellet
辐照燃料颗粒的数字放射自显影
- 批准号:
370568-2008 - 财政年份:2008
- 资助金额:
-- - 项目类别:
University Undergraduate Student Research Awards