Vilim F S, Aarnisalo A A, Nieminen M L, Lintunen M, Karlstedt K, Kontinen V K, Kalso E, States B, Panula P, Ziff E
Howard Hughes Medical Institute, Department of Biochemistry, New York University Medical Center, New York, New York, USA.
Mol Pharmacol. 1999 May;55(5):804-11.
Neuropeptides FF (NPFF), AF (NPAF), and SF (NPSF) are homologous amidated peptides that were originally identified on the basis of similarity to the molluscan neuropeptide FMRF-amide. They have been hypothesized to have wide-ranging functions in the mammalian central nervous system, including pain modulation, opiate function, cardiovascular regulation, and neuroendocrine function. We have cloned the NPFF gene from human, bovine, rat, and mouse, and show that the precursor mRNA encodes for all three of the biochemically identified peptides (NPFF, NPAF, and NPSF). We demonstrate that NPFF precursor mRNA expression by Northern analysis and map sites of expression by in situ hybridization. We confirm the validity of the in situ hybridization by showing that its distribution in the brain and spinal cord matches the distribution of NPFF and NPSF immunoreactivity. We go on to show that the mRNA levels (as measured by in situ hybridization) in the spinal cord can be up-regulated by a model for inflammatory pain (carrageenan injection), but not by a model for neuropathic pain (lumbar nerve ligation). Our results confirm the evolutionary conservation of NPFF, NPAF, and NPSF neuropeptide expression in mammalian brain. They also provide a context for the interpretation of the pain-sensitizing effects of injections of these peptides that have been previously reported. Our results support a model for the role of these peptides in pain regulation at the level of the spinal cord.
神经肽FF（NPFF）、AF（NPAF）和SF（NPSF）是同源酰胺化肽，最初是根据与软体动物神经肽FMRF酰胺的相似性鉴定出来的。据推测，它们在哺乳动物中枢神经系统中具有广泛的功能，包括疼痛调节、阿片样物质功能、心血管调节和神经内分泌功能。我们已经从人、牛、大鼠和小鼠中克隆了NPFF基因，并表明前体mRNA编码所有三种经生化鉴定的肽（NPFF、NPAF和NPSF）。我们通过Northern分析证明了NPFF前体mRNA的表达，并通过原位杂交确定了表达位点。我们通过显示其在脑和脊髓中的分布与NPFF和NPSF免疫反应性的分布相匹配，证实了原位杂交的有效性。我们接着表明，脊髓中的mRNA水平（通过原位杂交测量）可被炎症性疼痛模型（角叉菜胶注射）上调，但不能被神经性疼痛模型（腰神经结扎）上调。我们的结果证实了NPFF、NPAF和NPSF神经肽在哺乳动物脑中表达的进化保守性。它们还为解释先前报道的这些肽注射的疼痛致敏作用提供了背景。我们的结果支持了这些肽在脊髓水平的疼痛调节中作用的模型。
