van Golen F A, Li K W, de Lange R P, Jespersen S, Geraerts W P
Graduate School Neurosciences Amsterdam, Research Institute Neurosciences Vrije Universiteit, Faculty of Biology, The Netherlands.
J Biol Chem. 1995 Nov 24;270(47):28487-93. doi: 10.1074/jbc.270.47.28487.
An innovative method, direct peptide profiling of small samples of nervous tissue by matrix-assisted laser desorption ionization mass spectrometry, in combination with peptide characterization, immunocytochemistry in conjunction with specific neuronal labeling by backfilling of the penis nerve, and bioassay of peptides was used to study the intrinsic neuronal expression patterns of distinct sets of related FMRFa peptides and their significance for the organization of male copulation behavior in the mollusk, Lymnaea stagnalis. Previous studies indicate that the sets of FMRFa-related and GDPFLRFa-related peptides are encoded by two alternatively spliced transcripts of the single FMRFa gene. Direct mass spectrometry revealed that both FMRFa-related and GDPFLRFa-related peptides are present in the penis nerve, the sole nerve that innervates the penis complex. Accordingly, authentic FMRFa, GDPFLRFa, and related peptides were purified from the penis complex. The loci of synthesis of FMRFa and related peptides could be traced to the right cerebral ventral lobe, those of GDPFLRFa and related peptides to the B group neurons in the right parietal ganglion and to a few unidentified neurons in the right pleural ganglion. Notwithstanding their related structures, the two sets of peptides have distinctly different actions on the penis retractor muscle.
一种创新方法，即通过基质辅助激光解吸电离质谱对神经组织小样本进行直接肽谱分析，并结合肽表征、免疫细胞化学以及通过阴茎神经回填进行特定神经元标记，以及肽的生物测定，用于研究相关FMRFa肽不同组的内在神经元表达模式及其对软体动物椎实螺雄性交配行为组织的意义。先前的研究表明，FMRFa相关肽组和GDPFLRFa相关肽组由单个FMRFa基因的两个可变剪接转录本编码。直接质谱分析显示，FMRFa相关肽和GDPFLRFa相关肽都存在于阴茎神经中，阴茎神经是支配阴茎复合体的唯一神经。因此，从阴茎复合体中纯化出了 authentic FMRFa、GDPFLRFa及相关肽。FMRFa及相关肽的合成位点可追溯到右大脑腹叶，GDPFLRFa及相关肽的合成位点可追溯到右顶神经节中的B组神经元以及右胸膜神经节中的一些未鉴定神经元。尽管这两组肽结构相关，但它们对阴茎收缩肌的作用却明显不同。
