Study of crustacean humoral substances released during agressive behavior

我之前发现，当龙虾受到压力刺激时，美国龙虾（Homarus americanusu）会向血液中释放未知的压力物质，这是用微透析- hlpc技术分析血液成分时发现的。例如，在一种多刺龙虾（Panuliurus japonicus）甚至人类身上都检测到了这种特殊物质。目前的检测系统分析的不是单胺类物质。未知峰不是尖峰。它是广泛的，仅在施加应力刺激后的前5分钟收集的微透析液中发现。用高效液相色谱法测定了甲壳类动物的各种候选肽激素，并对其进行了检测。其中包括红色素浓缩激素、proctolin、胰岛素样肽等。结果发现，只有proctolin是主要候选物质。采用TOFMAS（飞行时间质谱法）分析小龙虾（龙虾替代品）心脏窦血中是否含有proctolin。当溶液浓度为1 mM时，TOFMAS可检测到标准的直肠球蛋白，但血液中未检出直肠球蛋白。血液中Proctolin浓度可能较低，低于质谱法的检测限。为了将未知物质定性为proctolin，有必要在分析仪测试前进行分离和浓缩。

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.