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

神经肽FF(FlFQPQRFamide)最初是从牛体内分离得到的。 并被发现具有阿片类药物的调节活性。为了获得更好的 对阿片类药物调节作用机制的认识 对于NPFF，我们开始了一项分离ProNPFF特异性基因的研究。 最初，使用从NPFF推导出的简并的27聚体寡核苷酸 筛选大鼠下丘脑cDNA文库，发现难度较大。 为了便于克隆，可能需要有多个 寡核苷酸探针(来源于NPFF前体的不同部分 蛋白质)。因此，在本研究中，NPF样多肽与 从大鼠中枢神经系统中分离出NPFF，纯化后的多肽 特色化的。其中一种纯化的多肽被鉴定为 SLAAPQRFAMIDE。通过测序确定了多肽序列，并对其进行了鉴定。 NPFF抗血清的特异性。该多肽序列进一步 通过比较内源性多肽和合成的 反相高效液相色谱法中的SlaapqRFamide。合成肽 SLAAPQRFamide被发现与NPFF受体具有高亲和力 这表明NPFF和SLAAPQRFamide可能具有相同的前体。我们 能够对另一种多肽进行部分测序，这种多肽 发现其n-末端有Pro-Ser-Leu-Ala。这些附加内容 多肽序列将用于制备额外的寡核苷酸 辅助分离ProNPFF特异性cDNAs的探针NPFF是高度 定位于离散的中枢自主神经区，包括丘脑核团 孤束和臂旁外侧核。这个 NPFF在心血管自主神经调节中的可能作用 通过检测这些区域的激活来研究这些区域的反应 NTS NPFF神经元，用Fos免疫反应。结果表明， NTS中的NPFF神经元对特定的心血管刺激作出反应 由低血压产生，但对其他应激刺激，如 中央产生的鸦片类药物戒断。

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.