| Chinese mitten crab, Eriocheir sinensis, commonly known as crab. It is a unique freshwater crabs in China and has become one of the major global economic species because of its delicious and nutritious in recent years. Healthy growth is closely related to the digestive physiology of crustaceans, but the study about the digestive system is few. Serotonin( 5-hydroxytryptamine, 5-HT) is a kind of biogenic amine that may regulate the digestion, absorption and metabolism of crustaceans. 5-HT is widely distributed in the digestive tract of crustaceans and closely related to the digestive function. It is important to research that how 5-HT functions in the digestive system. In order to study that how 5- HT plays a role in the process of crustaceans activities life. This experiment studied the distribution of the 5-HT related receptors and the neurotransmitter related to 5-HT through adopts the method of immunohistochemistry and if the content of 5-HT changes in different tissues in 24 hours. By injection of 5-HT, we observe whether that it influencese to food running time. Cajal interstitial cells(ICC) is a special kind of mesenchymal cells, that distributes in the digestive system, participates in the activities of the smooth muscle of the pacemaker and takes parts in regulation of the neurotransmitter, promotes the spread of electrical activity. Eriocheir sinensis can survive with lack of food for a long time. Hunger can affect digestive system, till now, the research about the change of crustaceans of the digestive system after hunger is more and more, but if hunger will change ICC hasn’t been research. This study observed the change of the ICC after hunger for the first time, to further investigate the the impact of hunger of crustaceans physiological state.This research mainly divided into three parts, respectively is:1. Study on the content of 5-HT of eriocheir sinensis mainly organization(eye stalkand brain, thoracic ganglia) and the study of the relevant receptors of the gut 1) This experiment by liquid phase measurement determinated the content of 5-HT in eriocheir sinensis serum, hepatopancreas, gut and thoracic ganglia, and by ELISA method determinated the content of 5-HT in eye stalk and the brain, the results are as follows: a Within 24 hours, as time goes on, a significant reducted in the content of 5- HT in serum; b Within 24 hours, as time changes, the content of 5-HT in the brain presented down rising trend. And the same time inside the content of 5-HT in the eye stalk is lower than the levels in the brain. c In the same feeding status,the content of 5-HT in hepatopancreas is higher than the other tissues. d In different feeding status, the content of 5-HT in hepatopancreas has significant difference, at the same time, the content in the other tissues don’t have significant difference. 2) The hepatopancreas, by immunohistochemical method to determinate the distribution of 5- HT1 A, 5- HT2 A, ACHE, NOS1 and NOS3 in the gut, the results are as follows: a 5-HT1 A can be detected by immunolization in the hindgut ampulla, and was widely distributed in peripheral connective tissue. 5-HT2 A can be detected by immunolization in the whole gut and the hepatopancreas, in the midgut, it is mainly distributed in the nucleus of mucosal epithelial cells and adventitia. In the hindgut ampulla, and was widely distributed in peripheral connective tissue and the nucleus of mucosal epithelial. In the hindgut,5-HT2 A is mainly found in the nucleus of mucosal epithelial cells and adventitia. In the hepatopancreas,it is mainly found in the four cell nuclei(E- cells, B cells, R- cells and F- cells), the F- also have positive distribution in the cytoplasm of cells. b ACHE can be detected by immunolization in the whole gut. In the midgut, it is mainly distributed in the nucleus of mucosal epithelial cells and adventitia. In the hindgut ampulla, was widely distributed in peripheral connective tissue and the nucleus of mucosal epithelial. In the hindgut, it is mainly found in the nucleus of mucosal epithelial cells and adventitia.c NOS1 and NOS3 can be detected by immunolization in the whole gut. In the midgut, it is mainly distributed in the nucleus of adventitia. In the hindgut ampulla, was widely distributed in peripheral connective tissue. In the hindgut,it is mainly found in the adventitia. 3) By argentation to determinate the distribution of 5-HT cells in the intestinal a The study appraisal eriocheir sinensis by the altered De Grandi silver nitrate dyeing method and Grimelius silver nitrate method, silver cells were not found in the intestines, by Grimelius silver nitrate method silver cells were found in groups or single silver cells the hepatopancreas.2 The effect of 5- HT of eriocheir sinensis to food running time, and the content of ACH and NO in the intestines 1) This experiment has uesed the camera to observe by injection of 5- HT and its agonists and antagonists of eriocheir sinensis to get the food running time. a This study found that 5- HT can shorten food running time in eriocheir sinensis in the gastrointestinal tract. b Eye stalk after resection, 5-HT can extended the food eriocheir sinensis in the gastrointestinal tract. c a- methyl- 5- HT(5- HT2 A agonist)can be further shorten the time of the food in the gastrointestinal tract. 2) In this experiment by liquid phase method at different feeding condition after 5-HT injection to detect the change of eriocheir sinensis hepatopancreas, intestines, and thoracic ganglia, method by Nanjing jiancheng under different feeding status after 5- HT injection to detect of the change of the content of ACH and NO in the gut. a After injection of 5- HT,the content of 5-HT increased in intestinal of eriocheir sinensis after defecation. b After injection of 5- HT, 5-HT2 A receptor has reduced in the hindgut ampulla and the hindgut after defecation c Exogenous 5- HT can reduce the content of ACH and NO in the feeding, especially the content of NO.3 Preliminary study on morphology of ICC that closely related to intestinalcontraction of the intestinal stromal 1) By transmission electron microscope to observe eriocheir sinensis intestinal structure and ICC form, the results are as follows: a A preliminary observation on eriocheir sinensis intestinal tissue structure, for the mucosa from inner to outer, submucosa, muscular layer and adventitia, observe the epithelial cells, granular cell and the electron structure of the ICC; 2) Under the condition of hunger, through ZIO method and SEM method to observe the change of eriocheir sinensis ICC cells in the gut, and the results are as follows: a By ZIO, along with a longer duration of hunger, the distribution of ICC on intestinal submucosa is on the rise. b After starvation, intestinal epithelial cells change from a single nucleus to multiple nuclei; The content of organelles in ICC decreased significantly in hindgut; with the starvation time passed by, muscles apparent to melt. |
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