NEUROPEPTIDE FF (FLFQPQRF-NH2) IN BRAIN AND SPINAL CORD

大脑和脊髓中的神经肽 FF (FLFQPQRF-NH2)

• 批准号: 5203692
• 负责人: H-Y T YANG
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF MENTAL HEALTH
• 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/5203692
• 关键词: brain; brain mapping; drug addiction; drug interactions; drug tolerance; endogenous opioid; experimental brain lesion; high performance liquid chromatography; laboratory rat; molecular cloning; neuropeptide receptor; neuropeptides; neuropharmacology; neurotransmitters; opioid receptor; protein metabolism; protein sequence; protein structure function; spinal cord

Neuropeptide FF (FLFQPQRFamide) was originally isolated from the bovine brain and found to have opiate modulating activity. To gain a better understanding of the mechanisms involved in the opioid modulatory role of NPFF, a study was initiated to isolate proNPFF specific cDNA. Initially, degenerate 27-mer oligonucleotides deduced from NPFF was used to screen rat hypothalamic cDNA library and was found to be difficult. To facilitate the cloning, it may be desirable to have multiple oligonucleotide probes (deduced from different parts of NPFF precursor protein). Thus in this study, NPFF-like peptides which are distinct from NPFF were isolated from the rat CNS and the purified peptides were then characterized. One of the purified peptide was characterized as SLAAPQRFamide. The peptide sequence was determined by sequencing and the specificity of the NPFF antiserum. The peptide sequence was further confirmed by comparing the endogenous peptide with the syntesized SLAAPQRFamide in a reverse phase HPLC. The synthetic peptide SLAAPQRFamide was found to bind to the NPFF receptor with a high affinity indicating that NPFF and SLAAPQRFamide may have the same precursor. We were able to partially sequence one other peptide and this peptide was found to have Pro-Ser-Leu-Ala at its n-terminal end. These additional peptide sequences will be used to prepare additional oligonucleotide probes to aid in the isolation of proNPFF specific cDNA. NPFF is highly localized within discrete central autonomic regions including nucleus of the solitary tract (NTS) and the lateral parabrachial nucleus. The possible role of NPFF in the autonomic regulation of cardiovascular responses in these regions was studied by examining the activation of NTS NPFF neurons, using Fos immunoreactivity. The results indicated that NPFF neurons in the NTS respond to specific cardiovascular stimulation produced by hypotension, but not to other stressful stimuli such as centrally-generated opiate withdrawal.

神经肽FF（FLIPQRF酰胺）最初从牛中分离， 大脑，并发现有阿片调节活性。更好地 了解阿片调节作用的机制 为了分离proNPFF特异性cDNA，开始了一项研究。 最初，使用从NPFF推导的简并27-mer寡核苷酸 筛选大鼠下丘脑cDNA文库，发现困难。 为了便于克隆，可能期望具有多个 寡核苷酸探针（从NPFF前体的不同部分推导 蛋白质）。因此，在这项研究中，NPFF样肽不同于 从大鼠CNS中分离NPFF，然后将纯化的肽段 表征了纯化的肽之一被表征为 SLAAPQRF酰胺。通过测序确定肽序列，并将肽序列与肽序列进行比较。 NPFF抗血清的特异性。肽序列进一步被 通过比较内源性肽和合成的肽证实了这一点。 SLAAPQRF酰胺在反相HPLC中。合成肽 发现SLAAPQRF酰胺以高亲和力结合NPFF受体 表明NPFF和SLAAPQRF酰胺可能具有相同的前体。我们 能够对另一种肽进行部分测序， 发现在其N-末端具有Pro-Ser-Leu-Ala。这些额外 肽序列将用于制备另外的寡核苷酸 探针以帮助分离proNPFF特异性cDNA。 NPFF高度 位于不连续的中央自主神经区域内，包括 孤束（NTS）和臂旁外侧核。的 NPFF在心血管自主调节中的可能作用 在这些地区的反应进行了研究，通过检查激活 NTS NPFF神经元，使用Fos免疫反应性。结果表明 NTS中的NPFF神经元对特定的心血管刺激作出反应 由低血压引起，而不是由其他应激刺激引起 等 中枢性阿片类药物戒断反应