Expression Of MAPK In The Gynogenetic Diploids Crucian Carp

Mitogen-activatedproteinkinases(MARK) signaling pathway plays an important role in gene expression, proliferation, differentiation, apoptosis, metabolism and other physiological process of eukaryotic cells. Different ploidy commercial fishes have been successfully developed by the technology of intersectional crossing and gynogenesis. Unreduced gamete generated from the diploid gynogenetic progeny and the formations of unreduced gametes might be contributed by endoreduplication and fusion of germ cells. To study the generation of unreduced gamete, this experiment used the diploid gynogenetic progeny of allotetraploid hybrids as materials, investigated the expression profile of ERK(extracellular regulated protein kinases) and JNK(c-Jun N-terminal kinase) at diploid gvnogenetic progeny gonad early development. This work lays a foundation of further study on molecular mechanism regulation of the generation process of unreduced gamete. The main points of this dissertation are as follows:(1) According to the erk1 genes cDNA sequence in known species of zebrafish, cloned the intermediate fragment sequence of erk1 genes in red crucian carp, triploid crucian carp, allotetraploid hybrids and diploid gynogenetic progeny. The intermediate sequence of 709 bp have been obtained and the results indicate that the diploid gynogenetic progeny erk1 genes identities to diploid red carp, triploid crucian carp, allotetraploid hybrids, zebrafish erk1 genes in intermediate fragment sequence were 99.7%、99.7%、96.3%、91.9%, respectively.(2) Detected expression analysis of JNK, p-JNK, ERK in gonadal tissues with diploid gynogenetic progeny and different ploidy level fish by used the technology of Western-blot. Results showed that JNK and ERK had high amount expression in fish gonad tissues, but both of JNK and ERK expression didn’t existed significant differences in the ovary and testis with diploid gynogenetic progeny and different ploidy level fish. It was worthy of note that the p-JNK expression in diploid gynogenetic progeny gonad tissues was much higher than what was in triploid and tetraploid’s ovary.(3) The diploid gynogenetic progeny early gonad tissues observation showed that 6-10 months fish gonadal in the stage of I ovary development, and spermatogonium has a vigorous division in 8-10 months diploid gynogenetic progeny. Western-blot have been used in detected JNK and ERK expression, the results indicated that JNK expression in 10 months diploid gynogenetic progeny early gonad tissues were higher than that in 6 and 8 months, and ERK expression had a downward trend in 6, 8, 10 months diploid gynogenetic progeny early gonad tissues.(4) Immunohistochemical has been used to detect the expression of JNK and ERK in diploid gynogenetic progeny early gonad tissues. The results showed that both JNK and ERK were positive expressed in the spermatogonium of different months diploid gynogenetic progeny early gonad tissues. Compared with 6 and 8 months fish, the fish intensity of positive reaction of JNK were higher in10 months, but the results of ERK were reverse. The results were consistent with what were detected by Western-blot technology.In conclusion, MAPK signal transduction pathway, especially the function of JNK and ERK the diploid gynogenetic progeny gonad, has been researched in this paper. The results provided the evidence that JNK pathway may play an important role in the regulation of the diploid gynogenetic progeny produce unreduced gamete.