CAA: Relationship Between Fatty Acid-binding Proteins and Neurotransmitter Systems in Developing Brain

9509168赛尔纳这个项目解决了一种假设，即心脏亚型脂肪酸结合蛋白(H-FABP)的发育表达与大脑中神经递质系统的同时发育有关，即伽马酸。这一假说基于1)免疫组织化学对蛋白质的相似定位，2)当H-FABP也存在时大脑中特定路径/区域的已知分化时间，3)显示游离脂肪酸对GABA转运体和GABA-a受体的影响的数据，以及4)体外研究表明当外源H-FABP加入时突触体对GABA的吸收增强。这一假设将通过使用既定的方案，从新生小鼠小脑开发细胞培养系统来解决。将使用针对合成肽制备的抗血清和合成的寡核苷酸探针来检测培养物中H-FABP的表达。GABA转运体的存在将通过测量培养物对~3H-GABA的摄取来验证。接下来，我们将研究外源添加脂肪酸对H-FABP水平和GABA摄取率的影响。最后，在培养物中加入反义寡核苷酸以抑制H-FABP的表达，并确定对GABA摄取的影响。本提案中描述的活动旨在获得足够的试点数据，以保证提交一项重要的赠款提案。***

9509168 Sellner This project addresses the hypothesis that the developmental expression of the heart subtype of fatty acid-binding protein (H- FABP) is related to the simultaneous development of neurotransmitter systems in the brain, namely GABA. The hypothesis is based on 1) similar localization of the proteins by immunohistochemistry, 2) the documented timing of differentiation of particular pathways/regions in the brain at a time when H-FABP is also present, 3) data showing effects of free fatty acids on both the GABA transporter and the GABA-a receptor, and 4) in vitro studies showing an enhanced uptake of GABA by synaptosomes when exogenous H-FABP is added. The hypothesis will be addressed by developing a cell culture system from neonatal mouse cerebellum, using established protocols. The cultures will be examined for their expression of H-FABP using antiserum prepared against synthetic peptides and using synthetic oligonucleotide probes. The presence of the GABA transporter will be verified by measuring 3H- GABA uptake by the cultures. Next, the efdects of exogenously added fatty acids on the levels of H-FABP and on rates of GABA uptake will be studied. Finally, antisense oligonucleotides will be added to the cultures to inhibit the expression of H-FABP, and effects on GABA uptake will be determined. The activities described in this proposal are designed to obtain sufficient pilot data to warrant submission of a major grant proposal. ***