Study for the effects of neuroactive peptides newly isolated

新分离的神经活性肽的作用研究

• 批准号: 04044075
• 负责人: TAKEUCHI Hiroshi
• 金额: $ 3.07万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04044075/
• 关键词: Neuroactive peptides; Achatin-I; D-Amino acid residue; Neurotransmitter; Neuromodulator; Intracellular signal transduction system; Molluscan giant neurones; African giant snail (Achatina fulica Ferussac); 神経ペプチド; 興奮性神経伝達物質; 巨大神経細胞; 軟体動物; フィリピン

This year, we studied comprehensively effects of biologically-active peptides on the identifiable giant neurones of an African giant snail (Achatina fulica Ferussac) by the electrophysiological methods. The Achatina giant neurones were insensitive to the mammalian neurotransmitter peptides, such as substance P,Met-enkephalin, neurotensin, etc., except for oxytocin. On the other hand, these neurones were sensitive to the neuroactive peptides isolated from invertebrates. These findings were different from those of small molecule putative neurotransmitters, such as dopamine, serotonin and GABA.With these results, it was considered that one peptide neurotransmitter acts on the neurones of the less extent of animal species than the small molecule putative neurotransmitter. Achatina giant neurones were excited by achatin-I (Gly-D-Phe-Ala-Asp), isolated from Achatina ganglia by the collaboration of us, Suntory Institute for Bioorganic Research and University of Santo Tomas. APGW-amide (Ala-Pr … More o-Gly-Trp-NH_2), isolated also from Achatina ganglia by us, were inhibited Achatina giant neurones. These two peptides were proposed to be neurotransmitters for Achatina neurones. Besides, we demonstrated that Achatina cardio-excitatory peptide-1 (ACEP-1) (Ser-Gly-Gln-Ser-Trp-Arg-Pro-Gln-Gly-Arg-Phe-NH_2) isolated from Achatina ganglia by another group showed excitatory effects on Achatina giant neurones, and that fulicin (Phe-D-Asn-Gln-Phe-Val-NH_2) isolated from Achatina ganglia by another group had inhibitory effects on these neurones. We demonstrated also that FMRFFamide (Phe-Met-Arg-Phe-NH_2) isolated from the clam ganglia, AF1 (Lys-Asn-Glu-Phe-Ile-Arg-phe-NH_2), a related compound of FMRFamide isolated from an Ascaris, and [Ser^2]-Mytilus inhibitory peptide ([Ser^2]-MIP) (Gly-Ser-Pro-Met-Phe-Val-NH_2) isolated from Mytilus, had inhibitory effects on these neurones.The intracellular signal transduction systems for the excitatory effects of achatin-I and the inhibitory effects of fulicin were studied electrophysiologically using the inhibitors for these systems. H-89 (protein kinase (PK) A inhibitor) and W-7 (calmodulin inhibitor) inhibited the achatin-I effects, and IBMX (cyclic AMP phosphodiesterase inhibitor) facilitated the same effects, indicating that achatin-I acted via cyclic AMP-PKA system, and calmodulin participated in this system. On the other hand, KT-5823 (PKG inhibitor), calphostin C (PKC inhibitor) and W-7 suppressed the inhibitory effects of fulicin, indicating that fulicin acted via cyclic GMP-PKG system and/or IP_3 and DG-PKC systems, and calmodulin participated in these systems. IBMX suppressed the inhibitory effects, the mechanism of which cannot be explained at present.We demonstrated that APGW-amide, as well as achatin-I,acted not only as a neurotransmitter but also as a neuromodulator for Achatina neurones. Less

今年，我们通过电生理学方法综合研究了生物活性肽对非洲巨蜗牛（Achatina fulica Ferussac）可识别巨型神经元的影响。除催产素外，Achatina巨神经元对P物质、蛋氨酸脑啡肽、神经降压素等哺乳动物神经递质肽不敏感。另一方面，这些神经元对从无脊椎动物中分离出的神经活性肽敏感。这些发现与小分子推定神经递质如多巴胺、血清素和GABA的结果不同。根据这些结果，人们认为一种肽神经递质作用于比小分子推定神经递质更小范围的动物物种的神经元。乙酰胆碱酯酶巨神经元被我们、三得利生物有机研究所和圣托马斯大学合作从乙酰胆碱酯神经节中分离出来的乙酰胆碱酯酶 I (Gly-D-Phe-Ala-Asp) 激发。 APGW-酰胺（Ala-Pr … 更多 o-Gly-Trp-NH_2），也是我们从 Achatina 神经节中分离出来的，可以抑制 Achatina 巨神经元。这两种肽被认为是蜗牛神经元的神经递质。此外，我们还证明了另一组从Achatina神经节中分离出的Achatina心脏兴奋肽-1（ACEP-1）（Ser-Gly-Gln-Ser-Trp-Arg-Pro-Gln-Gly-Arg-Phe-NH_2）对Achatina巨神经元具有兴奋作用，并且从Achatina神经节中分离出的fulicin（Phe-D-Asn-Gln-Phe-Val-NH_2） 另一组的Achatina神经节对这些神经元有抑制作用。我们还证明了从蛤神经节中分离出的 FMRFFamide (Phe-Met-Arg-Phe-NH_2)、从蛔虫中分离出的 FMRFamide 的相关化合物 AF1 (Lys-Asn-Glu-Phe-Ile-Arg-phe-NH_2) 和 [Ser^2]-Mytilus 抑制肽 ([Ser^2]-MIP) 从贻贝中分离出的（Gly-Ser-Pro-Met-Phe-Val-NH_2）对这些神经元具有抑制作用。使用这些系统的抑制剂，对细胞内信号转导系统的achatin-I的兴奋作用和fullicin的抑制作用进行了电生理学研究。 H-89（蛋白激酶（PK）A抑制剂）和W-7（钙调蛋白抑制剂）抑制achatin-I的作用，IBMX（环AMP磷酸二酯酶抑制剂）促进相同的作用，表明achatin-I通过环AMP-PKA系统发挥作用，钙调蛋白参与该系统。另一方面，KT-5823（PKG抑制剂）、calphostin C（PKC抑制剂）和W-7抑制了fulicin的抑制作用，表明fulicin通过环GMP-PKG系统和/或IP_3和DG-PKC系统发挥作用，钙调素参与了这些系统。 IBMX 抑制了这种抑制作用，其机制目前还无法解释。我们证明，APGW-酰胺以及阿契丁-I 不仅可以作为神经递质，而且可以作为阿契蒂纳神经元的神经调节剂。较少的

项目成果

TAKEUCHI,H.,Kim,K.H.,Liu,G.J.,YASUDAKAMATANI,Y.,Mi: "Achatin-I,an excitatory neurotransmitter having a D-phenylalanine residue of Achatina giant neurones" In″Invertebrate Neurobiology″,Akademiai Kiudo,Budapest,

TAKEUCHI,H.,Kim,K.H.,Liu,G.J.,YASUDAKAMATANI,Y.,Mi：“Achatin-I，一种兴奋性神经递质，具有 Achatina 巨型神经元的 D-苯丙氨酸残基”，《无脊椎动物神经生物学》，Akademiai Kiudo，布达佩斯，

Santos,D.E.,Takeuchi,H.: "Influence of the drugs for membrane excitability modification on the excitation caused by achatin-I" Comparative Biochemistry and Physiology.

Santos，D.E.，Takeuchi，H.：“膜兴奋性修饰药物对 achatin-I 引起的兴奋的影响”比较生物化学和生理学。

Santos D.E.and Takeuchi H.: "Influence of drugs for membrane excitability modification on the excitation caused by achatin-I." Comparative Biochemistry and Physiology. 105C. 185-188 (1993)

Santos D.E. 和 Takeuchi H.：“膜兴奋性修饰药物对 achatin-I 引起的兴奋的影响。”

Liu,G.J.,Takeuchi,H.: "Effects of cyclic AMP,cyclic GMP and IP_3 intracellularly injected into the identifiable Achatina giant neu.." Comparative Biochemistry and Physiology.

Liu，G.J.，Takeuchi，H.：“细胞内注射环 AMP、环 GMP 和 IP_3 到可识别的 Achatina 巨神经元中的效果..”比较生物化学和生理学。

Takeuchi H., Araki Y., Emaduddin M., Zhang W., Han X.Y., Salunga T.L.and Wong S.M.: "Identifiable giant neurones : their localizations in ganglia, axonal pathways and pharmacological features." Gen.Pharmacol.(in press).

Takeuchi H.、Araki Y.、Emaduddin M.、Zhang W.、Han X.Y.、Salunga T.L. 和 Wong S.M.：“可识别的巨型神经元：它们在神经节、轴突通路和药理学特征中的定位。”