Studies On Spermathophore Dissolution And Sperm Acrosin Activity Of Eriocheir Sinensis

This paper was based on the propagate problems in breed aquatics and research status quo about the Chinese mitten-handed crabs (Eriocheir Sinensis) at home and abroad. The research, which circumfusing the changing process that from Spermatophores to sperms, worked on the spermatheca in female, accessory gland in male and other aspects hailed from these. On the one hand, it can contribute to natural science research by offering basic theoretics material and on the other hand, it can help to make the freshwater economic resourch of our country developed and utilized better.1. The spermatheca, which is an important organ in Eriocheir Sinensis female procreation system, plays vital function on sperm storage, ovulation and fecundation activities. Using histological, histochemical and transmission electron microscope(TEM) technology, basic research was operated on the spermatheca of the mature female Eriocheir Sinensis in order to reveal the main structure of the spermatheca wall and the chemical inclusion of spermatheca cells.The spermatheca is devided into two parts: vesicle wall and vesicle lumen. From the outside in, vesicle wall contains: Connective tissue(CT), Layler of muscle(LM), basement membrane cell(BM) and Columnar epithelium cell(CEC). These parts are not existed in the same place and some position of the spermatheca own only one or two parts. So, vesicle wall of Eriocheir Sinensis spermatheca is typical transitional epidermal.Result of TEM showed: many microvilli(MV) existed on the end of CEC and several different kinds of lumen secretion(LS) existed in the vesicle lumen. The function of this structure is hypothesized playing dissimilar action in procreation process.Analysing the main structure and chemical inclusion of cells of spermatheca to make sure that the spermatheca does a contribution to depositing nutrition when crabs are mating and breeding, therefore we can speculate the source of nutrition of the sperms after mating. The apical excreting cell contains lots of carbohydrate , some protein and fat, especially, it excrete much more carbohydrate than any other cells. The spermathecal epithelium cells provide nurishment to sperms keeping metabolism in spermatheca, so it can be assumed the carbohydrate from apical excreting cells be the main nutrients of sperms in spermatheca, and the protein and fat excreted from epithelium cells may participate in enzymatic reaction of the sperm.2. Spermatophores of the Eriocheir sinensis, which appeared in Proximal vasa deferentia, throughing vasa deferentia, sperm bundles, accessory gland and ejaculatory duct, entered into spermatheca with semen and other male sex gland exudates. Short time after the Spermatophores entered the spermatheca, it dissoluted and existed as single sperm until combining with germ cell. The aim of this experiment is to find out more information about the process from spermatophore to single sperm.Using man-made seawater to wash and dilute Spermatophores of the Chinese mitten-handed crabs (Eriocheir sinensis) to made the concentration of which was 1500/ ml. Three important reproductive apparatus: accessory gland, ovary and spermatheca were made to homogenization, and four different protein solutions were distilled from these three homogenate liquids and spermatic in the condition of refrigerant centrifugal machine, separately.Method of Coomassie Brilliant Blue was adopted to detect and balance the concentration of proteins to the same concentration. Five protein groups: contrast, spermatic, accessory gland, ovary and spermatheca were set up in the same condition and combined with Spermatophores. Put 30ul mixed reactive solution to slide in each group and at fixed time and taken count of the survival number of integrated Spermatophores. Results showed that four proteins: contrast, spermatic, overy and spermatheca have no prominent impact on dissolution of Spermatophores but accessory gland protein has marked function to accelerate the Spermatophores dissolve. Thus we can concluded that accessory gland protein play a vital role in dissolution of spermatophora.3. The accessory gland of Eriocheir sinensis located between sperm bundles and ejaculatory duct, and these three organ are communicate. The accessory gland is white, distribute on both side of the crab symmetrically. It has been proved that accessorygland protein make function on spermatophore dissolution.On the foundation that accessory gland protein can dissolute the spermatophore, by using scanning electron microscopy(SEM) and protein electrophoresis technology, more information about which kind of protein it is and where it situates can be researched futher.Using SEM sample 2 and 3 digesting Spermatophores reveals that the protein that can dissolute spermatophore exists in the accessory gland, but it is not excreted into the vesicle lumen at peacetime. Rusult of SEM showed: compared to integrate inclusion, the big vesicles become small and losing its regular rotundity, and what's more, the number of small vesicles is huge. The result of the protein electrophoresis showed: the number of proteins in small vesicles is remarkable more than that in big vesicles. It can be concluded that the protein in small vesicles of accessory gland play vital or full function on spermatophore dissolution.4. Acrosin, a sperm-specific arosomal proteinase, has an essential role in the fertilization process and is thought to be important in the acrosome reaction, sperm-zona binding and zona penetration. The spermatozoa acrosin activity and the accessory gland acrosin activity of Eriocheir sinensis are measured with a simplified clinical assay of total sperm acrosin activity. This method is based on the measurement of amidase activity of sperm suspensions. The acrosin activity assay has been used in many animals. Results of these studies indicated that use of this assay method could provide valuable information concerning semen quality.The accessory gland acrosin activity of Eriocheir sinensis is about 3.216μIU·10^-6 and 2.232μIU·10^-6,which is remarkable lower than the spermatheca gland acrosin activity: 239.090μIU·10^-6 and 219.297μIU·10^-6. It can be concluded that although the protein of accessory gland can dissolute the spermatophore, it have less relation to the high acrosin activity in spermatheca. The factor that effect the acrosin activity should come from spermatheca itself. Former experiment proves there are abundant secretion in spermatheca and these secretion maybe have some relation with improvement of the acrosin activity. This result indicates that the clinical assay of acrosin activity can be used for crab spermatozoa to evaluate the quality of crab semen.