Study Of ChM-I And VCAM-1 Gene Sequential Expression In The Tibial Dyschondroplasia Of Broiler

Tibial dyschondroplasia (TD), is a common disease of skeletal in broiler, the characteristic is accumulat growth plate cartilage into metaphyseal region of the tibiotarsus so that formated neither vascularize nor mineralize cartilage. The development of TD is involved in many factors such as genotype, the calcium/phosphorus ratio and acid-base balance of the diet, the TD has made huge loss of economy in poultry industry ,although scientists have made a lot of research about the case of TD in recent years, the detailes etiologic process of TD are not completely understand.ChM-I (chondromodulin-I) was a cartilage-derived molecules found in recent years,the ChM-I can stimulateing the growth of cartilage cellsand and osteoblasts ,but it can inhibit vascular endothelial cell growth,so ChM-I play an important role in the process of avessels. VCAM-1 is an important adhesion molecules,which is concerned with immune cell adhesion and migration,and participate in the growth of hematopoietic cells, VCAM-1 can assist the development and adhesion of hematopoietic cells located in the micro-environment,participate in hematopoietic cells and stromal cells in the formation of aggregates,by the way ,it also can participate in hematopoietic cell homing and migration, so VCAM-1 in cartilage differentiation and the process of vessels play an important role.Application of suppression subtractive hybridization (SSH) technology have selected 10 different expressed genes before this research, including the ChM-I and VCAM-1, but we have not know about the difference broilers expression at different stages of TD,for this reasone, this study use realtime q-PCR,situ hybridization and immunohistochemistry respectively, research ChM-I and VCAM-1 gene expression differences in timing at gene and protein levels on the occurrence of TD .The obtained results are as follows:The results of realtime q-PCR and in situ hybridization display that the ChM-I gene express upward or downward at different times of TD, Specific performance is show that: ChM-I gene expressed upward at the 1 st and 4th samplings and downward at the 2nd and 3rd samplings when give animal feeds with the thiram; VCAM-1 gene express downward at the 1st, 2nd , 3rd and 4th samplings.The ChM-I of express vector PGEX-KG system was succesful found, the result of SDS-PAGE and Western blot indicated that the molecular weight of ChM-I was about 41 Kda,and exist in inclusive body. Uesing the ChM-I expressed in the E.coli as antigen to immuntied New Zealand rabbit, rabbit anti-ChM-IIGg has been maded and purified. Rabbit anti-ChM-I IGg was used as the frist antibody in immunohistochemistry. The results show that the quantity of ChM-I has changed in different period in TD,but it is not very consistent with the results of realtime q-PCR and in situ hybridization.This study show that ChM-I and VCAM-1 both in gene level and in protein level are diversity , analysis of these changes and the reasons can provid new basis theory and ways for the pathogenesis study of TD.