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研究了生物活性宠物对非洲巨型蜗牛（Achatina funica Ferussac）的鉴定巨型神经元的彻底影响。 Achatina巨型神经元对哺乳动物神经递质的宠物不敏感，例如PESSS P，Met-Enkephalin，Neurotensin等，除催产素外。另一方面，这些神经元对从无脊椎动物分离的神经活性宠物敏感。这些发现与小分子假定的神经递质的发现不同，例如多巴胺，5-羟色胺和GABA。与这些结果相比，与小分子假性神经递质相比，一种辣椒神经递质作用于动物种类较小的神经元的神经元。 Achatin-I（Gly-D-Phe-Ala-Asp）激发了Achatina巨型神经元，该神经元被美国合作，圣诞节生物有机研究研究所和桑托·托马斯大学（University of Santo Tomas）从Achatina神经节中分离出来。 APGW-Amide（Ala-pr…更多的O-Gly-gly-trp-NH_2）也抑制了Achatina巨型神经元，也从Achatina神经节中分离出来。认为这两个宠物是Achatina神经元的神经递质。此外，我们证明了Achatina心脏优异肽-1（ACEP-1）（ser-gly-gln-ser-ser-ser-trp-arg-arg-arg-pro-gln-gly-gly-arg-arg-arg-phe-phe-nh_2）由另一组从阿帕蒂娜神经节中分离出来，显示了另一组由另一组从阿帕蒂娜神经节中分离出来的另一组的神经节对这些神经元有抑制作用。我们还证明了FMRFFAMIDE（phe-met-arg-phe-nh_2）从蛤laglia（Af1）（lys-asn-asn-asn-glu-phe-ile-arg-phe-nh_2）中分离出来（Gly-ser-pro-met-phe-val-nh_2）从mytilus分离出来，对这些神经元具有抑制作用。细胞内信号转导系统，用于achatin-i的兴奋性效应和fulicin by fulicin by fulicin by fulicin的抑制作用，并使用这些系统的抑制剂进行了抑制作用。 H-89（蛋白激酶（PK）抑制剂）和W-7（钙调蛋白抑制剂）抑制了achatin-I效应，IBMX（环状AMP磷酸二酯酶抑制剂）支持了相同的效果，表明achatin-i通过环状AMP-PKA系统和Calmododudulin Comented Comented Consecy cons and Achatin-I作用。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通过环状GMP-PKG系统和/或IP_3和DG-PKC系统以及Calmodulin参与这些系统。 IBMX抑制了抑制作用，目前无法解释其机制。我们证明，APGW-酰胺和achatin-I不仅充当神经递质，而且还作为achatina神经元的神经调节剂。

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.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。”

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。”

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。

Liu G.J.,Santos D.E.and Takeuchi H.: "Sensitivities of Achatina giant neurones to peptides isolated from Mollusca." Acta Biologica Hungarica. 44. 25-28 (1993)

Liu G.J.、Santos D.E. 和 Takeuchi H.：“Achatina 巨型神经元对从软体动物中分离出的肽的敏感性”。