Isolation And Characterization Of Genes Involved Before And After Puberty In Xiao Meishan Pig's Hypothalamus

At present, domestic and foreign scholars on a variety of animal-related puberty the regulator has done a wide range of research, explained the steroid hormones, growth factors, neurotransmitters, body fat and other metabolic signals of neurons in the hypothalamus of the impact of the release of GnRH. However, apart from the above mentioned factors on the regulation and control pigs puberty to start the adjustment, there are still unknown whether the impact of regulatory genes in pigs start puberty is unclear. The purpose of this study by RT-PCR, differential display PCR (DDRT-PCR), Reverse Northern, quantitative fluorescence molecular biology techniques, such as the beginning of the situation with a view to the unknown gene screening, identification, and their developmental stages in different physiological expression linked to changes in the law, to explore the situation of these genes in the early period of the role of the mechanism will be activated in order to enrich the mechanism of animal reproduction and breeding early maturing varieties and provide a theoretical basis for animal selection.hypothalamus samples were collected from 3 pigs aged new born(0d),puberty(70d),sexual maturity(150d).Two RNA pool groups from 9 pigs, each consisted of 3 samples, of new born,puberty (â… group) and sexual maturity,puberty(â…¡group), were selected to compare the differentially expressed genes between the groups. DDRT-PCR reaction was performed by using 100 primers combinations of 10 anchored primers coupled with 10 arbitrary primers. Twelve Expressed Sequence Tags (ESTs) were gained and sequenced. The sequences were compared with GenBank database. Four out of twelve EST sequences are known genes. They were designated as MS01(355 bp), MS02 (391 bp), MS03 (241 bp) and MS04 (281 bp). MS01 sequence shows high homology (98%) with pig Brain-specific angiogenesis inhibitor 3(bpai 3), GenBank ID was XM₀01925295. MS02 is high homologous with pig brain-type fatty acid-binding protein (93%), GenBank ID was DQ000160.MS03 is homologous to cloneCH242-158N8 from chromosome 13 of pig (86%) , GenBank ID was CU633348. MS04 is homologous with human Splice variant of sigma-1 (93%) GenBank ID was DQ644568. Other ESTs were no homologou sequences in GenBank and other Databases.Finally, Reverse Northern, real time PCR detection of the qualitative and quantitative level of gene expression. Quantitative results showed that: pig hypothalamus in three developmental stages, bpai mRNA expression level showed a downward trend, and new-born at the time of gene expression with puberty and sexual maturity compared to a significant difference (P <0.05) , the result suggested that the hypothalamus birth to sexual maturity in the development process, this factor involved for the early period of the current situation hypothalamic neurons pulsatile-release to provide access to security, plays a leading role in the rapid development pipeline. fabp7 mRNA in three different developmental stages of pig hypothalamus, the expression level of the trend was upward, the expression of no significant difference (P > 0.05). The result suggested that fatty acid regulator of local pig breeds in China in its expression with the increase in the growth of day and gradually increased accumulation of a process, the factor as a fatty acid receptor, and enhance the affinity of fatty acids and their derivatives, to promote fatty acid uptake, suggesting the factor to further increase the accumulation of body fat pig, and the further accumulation of fat a direct impact on the results of the nutritional status of pigs, when the pig fat accumulation reached puberty start when the required level of body fat, and coordination other regulatory factors involved pigs start puberty.