Study On The Relationship Between Intermusc?lar Bone Development And Mechanical Stress In Zebrafish

Intermusc?Lar bones(IBs),also known as intermusc?Lar bones,are located in the muscle space on both sides of the vertebrae and are formed by the ossification of the connective tissue of the muscle septum.Intermusc?Lar bone is common in lower true bone fish,and there are three types of intermusc?Lar bone in most carps: med?Llary arch bone,vertebral bone,and vein arch bone.In recent years,most studies on fish intermusc?Lar bone have focused on the description of the morphological structure and development of intermusc?Lar bone,and less research has been done on the molec?Lar mechanism of intermusc ? Lar bone development.In vertebrates,there is evidence that mechanical forces affect the induction,growth,and maintenance of bones.Previous studies have reported a correlation between the fish's own movement pattern and the order of intermusc?Lar bone development.For example,fish such as catfish and eel use the muscle contraction in the front of the body to drive the head to swing,and the muscles of the fish that move the body forward are ossified from the head to the back;The intermusc ? Lar bones of fishes such as the cephalocephalus that use tail swing to drive the body forward are ossified from back to front.Zebrafish movement begins early in the individual's development,and mechanical stresses generated during these behaviors may affect subsequent bone development.In mice,the expression of the spliceosome ?2?FosB of FosB leads to bone sclerosis,and the expression of the osteoblast marker gene Runx2 is also increased.In zebrafish,microgravity reduced FosB expression,while moderate overweight res?Lted in increased FosB expression.These studies point to a central network of FOS-Jun factor members.The ossification of the FosB gene,especially its spliceo,? 2 ? FosB vertebrates,was induced by mechanical stress.The res?Lts of studies on skeletal development in mammals have given us inspiration.Does ?FosB have a similar role in the development of intermusc ? Lar bone in fish? Therefore,using the model animal zebrafish,we explored the hypothesis that mechanical forces may eventually lead to the production of intermusc?Lar bone by modifying the molec?Lar pathways of mechanical responses.This paper explores whether zebrafish skeletal development is affected by movement patterns and whether the expression of FosB gene changes under the condition of changing mechanical stress,so as to link mechanical stress with related FosB genes and jointly explore the zebrafish bone development.This study primarily explored the reg ? Latory mechanism of zebrafish FosB gene on bone development by recording the zebrafish development process,changes in exercise patterns,gene expression in various periods,immunofluorescence,and gene knockout.The main findings are as follows:First.This study used osteoblast-specific transcription factor green transgenic fluorescent protein(Osterix GFP)zebrafish strain(cy25)to observe the skeletal development process,and recorded the development of ribs and interspinous muscles in zebrafish under normal living environment.Second.Observe zebrafish bone development with changing water flow rate.It was found that under stress,the development of interspinous muscles and scales of the zebrafish was three days earlier than that of the control,and the development of scales was two days earlier.Third.RNA was extracted from each stage of zebrafish bone development,and the expression levels of the five spliceosomes of the FosB gene were determined by fluorescent quantitative PCR at each stage of bone development.The expression level of spines began to rise again in the middle stage of development.The analysis of the expression of Fos B gene in zebrafish under stress revealed that the expression of spliceosome increased significantly with the increase of exercise.Fourth.Take the zebrafish in the middle stage of intermusc?Lar spine development,and find the expression position of FosB gene protein by immunofluorescence and tissue section immunofluorescence on zebrafish whole fish,which are basically concentrated around the bones and on the scales.Fifth.FosB gene was knocked out by gene knockout technology,and the development of the ribs of the deleted heterozygote was found to be missing.This suggests that FosB gene plays a certain role in zebrafish bone development.Sixth.Using overexpression-related research to explore the related conditions of skeletal development,it was found that after injection of the overexpression plasmid,zebrafish showed different degrees of deformity and co?Ld not develop into a complete individual.In summary,from the comparative study of skeletal development under normal and mechanical stress conditions in this study,it was found that mechanical stress significantly affects ossification of zebrafish intermusc?Lar bone.And by analyzing the expression of FosB spliceosome at various stages,the expression of FosB gene in the zebrafish bone was determined by immunofluorescence detection,and the FosB gene was determined by gene knockout to affect the development of zebrafish bone.This study explored the effects of FosB gene on zebrafish bone development from the above aspects,laying a foundation for fish bone development and the development of interspinous spines,and providing a theoretical basis for bone development of teleost fish.