Immunohistochemical-Localization Of Opioid Growth Factor In Organs Of Thais Clavigera And Its Effect On The Development Of Thais Clavigera

Objective: Opioid growth factor (OGF), as a cytokine, has inhibitory effects on cell proliferation, however, there is little report about the effect on growth and development of Invertebrates. By using immunohistochemical techniques, we investigated the distribution of OGF in adult organs of Thais Clavigera, the reproductive system organs in different periods, embryos and larvae at various early stages. We also studied the law of Thais Clavigera's self-reproduction, and the injection of OGF and naloxone (Nal) into the oocysts in its early development period. At last we aggregated and discussed the effect of OGF on Thais Clavigera's growth and development, providing a theoretical basis for artificial propagation of Thais Clavigera and providing reference of heoretical and experimental for the impact of Opioid growth factor in animals ontogeny.Methods:(1)Position experiment: Immunohistochemical SABC technique was used to detect OGF's loci in adult and larvae of Thais Clavigera. And the provision is that the tan loci express is strong positive, yellow loci express is positive, light yellow loci express is weakly positive.(2)Injection of OGF and naloxone experiment: recording daily length growth of the larvae and calculating it's variational ratio, using statistical software SPSS11.5 to deal with experimental results.(3)Gonadal development cycle experiment: obtaining experimental materials regularly, observing with eyes and microscope, and recording the experimental results(.4)Reproduction experiment: observing reproduction behavior of Thais Clavigera for one year, recording the experimental results.Results: Based on the above fundamental research, we got several results of experiment: (1) The results of immunohistochemical-localization in adult showed that OGF was widely distributed in organs of Thais Clavigera. Furthermore, the nervous system and digestive system had more OGF positive loci than others; (2) The results of immunohistochemical-localization in embryo and larva demonstrated that embryo of 2-cell stage, 4-cell stage, 8-cell stage and16-cell stage had more OGF positive loci than veliger of medium-term and late, while gastrula, trochophora and veliger of early stage had less OGF positive loci; (3) The results of injection of OGF and naloxone into oocysts indicated that OGF slowed down the growth speed of laval while naloxone accelerated it. Moreover both effects showed dose effect. (4) The results of gonad development experiment displayed that there was certain regularity variety in the development of spermary and ovarian: gonad development cycle could be divided into five stages based on structural characteristics of gonad and maturity degree of sperm and ovum: proliferating stage (from September to February ), growing stage (from March to April ), maturing stage (May), spawning stage (from June to July) and resting stage (August). (5) The results of immunohistochemical-localization in organs of gonad demonstrated that there was a certain regularity in the distribution of OGF in gonad organs: spermary, ovarian and prostate had more OGF positive loci in proliferating stage, growing stage and resting stage than that in maturing stage and spawning stage. However, albumen gland, oocyst gland and male genital had more OGF positive loci in maturing stage and spawning stage than that in proliferating stage, growing stage and resting stage; (6) The results of Thais Clavigera's reproductive biology illustrated that Thais Clavigera had only one reproductive season each year: from June to July, in addition, peak of reproduction was from mid-June to mid-July. Thais Clavigera spawn 2-3 times each reproductive season, and it's hatchability (98.3%) is higher than others;Conclusions: (1) OGF was widely distributed in organs of Thais Clavigera. And the regularity of OGF's distribution in larval development and experiment of injecting OGF and naloxone demonstrated that OGF had an important role on Thais Clavigera's Development. (2) The OGF's distribution was regularity in gonad of various developmental stages.