Study On The Structure And Function Of Male Accessory Gland Of Eriocheir Sinensis

1. Histology of the accessory gland of Eriocheir sinensisThis paper described the conformation and structure of the accessory gland in Eriocheir sinensis, which were studied with anatomy, morphological and tissue section. The accessory gland of Eriocheir sinensis opening from the wall of seminal vesicle, consisting of cords of gland, each gland was composed of mang dendritic pipes, assuming radiation. There were abundant of contents in the cavity of accessory gland. Development of the accessory glands were exhibited in the length of gland, the volume of the cavity and the number of dendritic pipes. Standard histological techniques revealed: the wall of accessory gland was consisted of connective tissue, muscular tissue, membrane of connective tissue and epithelial tissue; outermost of the wall was of connective tissue, the shape of cells was shuttle, nucleuses were large and oblate; and interior of connective tissue was a layer of annular muscular. But the epithelium was different in the unitary gland.2. Ultrastructure of the accessory gland of Eriocheir sinensisWe used the TEM and SEM to investigate the ultrastructure of the accessory gland of Eriocheir sinensis, the results show that: there were affluent in mitochondria in basal membrane of the epithelium. The epithelium was riched in rough endoplasmic reticulum, which consisted in the top of cells, around nucleolus and in the basal of cells. Ribosome was affluent in the cytoplasm. The dissociation of epithelium cells has large numbers of Golgi complex; there were many of vacuole with different shape and volume in the top of cells. There were abundant of contents in the cavity of accessory gland, include vesicle secretions of electron-dense structure, there were granule secretions of electron-dense and many small vesicle secretions of electron- lucent between them. There were also many spermaphores in the cavity of accessory gland, which were surrounded by a thin layer of the spermatophore wall. There were many dense vesicle secretions of electron- lucent and granule secretions of electron-dense between spermaphores.3. Primary studies of the accessory gland secretions in Eriocheir sinensisWe used the histochemistry to investigate the component and the function of accessory gland secretions, the results of epithelium cells of accessory gland show that: periodic acid schiff (PAS) was positive, there were many glucide in accessory gland secretions; Alcian-PAS was moderately positive, showed that there were less acid mucopolysaccharide in epithelium cells of accessory gland; the reaction of propylene glycol Sudan IV was positive, showed that the secretions also contented little lipid; the reaction of Ninhydrin-Schiff was intensely positive, the mainly component of secretion is protein. The mensuration of biochemical composition showed that: the content of protein in accessory gland secretions was more than others. The results of SDS-PAGE showed that: proteins between 66kDa and 97kDa were more expressed in accessory gland of mature male.4. Primary study of function the accessory gland in Eriocheir sinensisWe used A23187 to abduct the acrosomal reaction of sperm, the results showed that: sperm in accessory gland that brought acrosomal reaction were distinct more than that in seminal vesicle. The results of electrophoresis showed that: proteins of accessory gland near 20kDa, 31 kDa and 66 kDa were not expressed in seminal vesicle, especially proteins of 34kDa, 67kDa, 72.5kDa and 74kDa in accessory gland were riched than seminal vesicle and ejaculatory duct. The protein brand of ejaculatory duct almost involved protein brands of seminal vesicle and accessory gland. The results of microscope showed that: the structures of spermatophores in seminal vesicle were integrity, and the configuration and edge of spermatophore wall were distinctness, the semen was made up of dense vesicle substance. The structures of spermatophores in the cavity of accessory gland were laxated, and the spermatophore wall also easy to break. The content of accessory gland was consisted of large number of nummular vesicle substance with different size, big vesicle substance with different shape and many dissociative sperm. 5. Study of digest the spermatophore of Eriocheir sinensis with accessory gland proteinsWe take out the accessory gland and used Ca²⁺-FASW for homogenization, Lowry method mensurated the concentration of accessory gland proteins was 15mg/ml.The purpose of this paper was to find a suitable method of acquiring free sperms from spermatophores of Chinese mitten-handed crab under two factors with different levels including a series of temperature (17 °C, 22°C, 27°C, 32°C and 37 °C) ,different concentrations in accessory gland proteins solution (15mg/ml, 7.5mg/ml, 3.75 mg/ml, 1.875 mg/ml, 0.9375 mg/ml, 0.46875 mg/ml and 0.234375 mg/ml), which to digest the same passel of spermatophores. The results showed that: the viability of free sperm was especially high, and little sperm died in 6 hours; at the same temperature the time of digestion of spermatophores were inverse ratio with concentration of accessory gland proteins, but spermatophores would be difficult to digest when the concentration were too low. At 32°C and 15mg/ml the time of digest was shortest. Basing on this, we compared the accessory gland proteins with trypsinase digest method, the results of FCM showed that accessory gland proteins were better than trypsinase digest method, which had much less impurity.