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Study of crustacean humoral substances released during agressive behavior

甲壳动物攻击行为期间释放的体液物质的研究

基本信息

批准号：
14540633
负责人：
YAZAWA Toru
金额：
$ 2.11万
依托单位：
Tokyo Metropolitan University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14540633/
关键词：
crustacea blood stress hormone lobster mass peptide ペプチド HPLC イセエビ行動

项目摘要

I have previously found taht the American lobster, Homarus americanusu, releases unknown stress-substance into blood stream when the lobster receives stress stimulation when blood ingredients were analyzed with microdialysis-HLPC technique. For example, the specific substance was detected in a spiny lobster species Panuliurus japonicus, and even a human. It was not monoamine substances that was analyzed by the present detection system. The unknown peak was not sharp peak. It was broad and only found in the microdialysate collected during the first 5 min after applying the stress stimulation. Various candidate peptide hormones of crustacean animals were tested with HPLC whether or not any hormone can be detected with the present amine-HLPC. Those include red pigment concentrating hormone, proctolin, insulin-like peptide, and so on. As the results, only proctolin was found to be the primary candidate. The blood taken from the cardiac sinus of crayfish (replacement of lobster) was analyzed whether or not it contains proctolin using TOFMAS (time of flight mass spectrometry). Standard proctolin was detectable with TOFMAS when solution concentration was 1 mM. But, no proctolin was found in the blood. Proctolin concentration in the blood might be low and less than detection limit of the mass spectrometry. To characterize the unknown substance as proctolin, it is necessary to separate and concentrate before testing by the analyzer.

本研究采用微透析-高效液相色谱技术分析了美洲龙虾（Homarusamericanusu）的血液成分，发现龙虾在受到应激刺激时，会向血液中释放未知的应激物质。例如，在一种多刺龙虾物种Panuliurus paclas，甚至是人类身上都检测到了这种特定物质。本检测系统分析的不是单胺类物质。那未知的山峰，并不是尖锐的山峰。它是广泛的，仅在施加应力刺激后的前5分钟内收集的微透析液中发现。用高效液相色谱法检测甲壳动物的各种候选肽类激素，以确定本胺-高效液相色谱法是否能检测到任何激素。包括红色素浓集素、原胆固醇、胰岛素样肽等，结果发现原胆固醇是主要的候选者。使用TOFMAS（飞行时间质谱法）分析从小龙虾（龙虾的替代物）的心窦采集的血液是否含有直肠菌素。当溶液浓度为1 mM时，TOFMAS可检测到标准品直肠菌素，但血液中未检测到直肠菌素。血液中的Proctolin浓度可能较低，低于质谱法的检测限。为了将未知物质表征为proctolin，在通过分析仪进行检测之前，有必要进行分离和浓缩。