Study On The Seasonal Differences Of Testicular Submicrostructure And MCM7,Bcl-2,Bax,p21,p53 Gene Expression Of Artificially Domesticated Tree Shrews

Objective:To investigate whether there are seasonal differences in the microstructure and submicrostructure of the seminiferous tubules during the occurrence of artificially domesticated adult tree shrews;To investigate whether there are seasonal differences in the mRNA and protein expression of MCM7,Bax,Bcl-2,p53,and p21 genes during the spermatogenesis of artificially domesticated adult tree shrews.Methods:Using 12 groups of adult male tree shrew testis tissues of different months as materials,the microstructure of testis tissue was observed by HE staining technique;the submicroscopic structure of testis tissue was observed by transmission electron microscope;Immunohistochemical detection of MCM7,Bax,Bcl-2,p53 p21 protein and DNA-mRNA in situ hybridization detection of MCM7,Bax,Bcl-2,p53,p21 genes were performed on the paraffin tissue sections.Results:H.E staining results showed that in January,the artificially domesticated tree shrew testicle deformed sperm in the seminiferous tubules and the residual amount of sperm deformed and shed less.During the period from February to April,the number of deformed sperm and the amount of sperm deformed and shedding residues tend to increase month by month,large amounts of deformed sperm cells and shedding residues that almost filled the lumen From May to August.In September,the number of spermatogenic cells,deformed sperm and sperm deformed residues were significantly reduced compared with August.In October,secondary spermatocytes,round sperm cells and deformed sperm cells increased compared with September.From November to December,the situation is similar to that of January,and the number of deformed sperm cells and the shedding residues are less.Transmission electron microscopy results showed that in the 12 months of the year,The largest number of free sperm in the seminiferous tubules of artificially domesticated tree shrew testicles was from March to July.The number of free sperm in August was less than in March-July,but more than in September-December and January-February.Apoptosis of some spermatogenic cells was observed in the seminiferous tubules of artificially domesticated tree shrew testis tissues during August-December and January.shell-like nucleoli appeared occasionally in primary spermatocytes in tree shrew testes In January and February.Malformed sperm were observed in tree shrew testicles from January to April and October to December.Immunohistochemical results showed that MCM7 protein had immunopositive signal in the seminiferous tubules of the tree shrew testes in each months of the year,mainly expressed in spermatogonia and primary spermatocytes,and there is a seasonal difference in the rate of positive cells;Bcl-2 protein had no immunopositive signal in spermatogenic cells at all levels of the seminal tubules of the tree shrews in January,mainly in spermatogonia and primary spermatocytes from February to October,and mainly in spermatogonia from November to December With a small number of primary spermatocytes,and there is a seasonal difference in the rate of positive cells;Bax protein has immunopositive signals in spermatogenic cells at all levels of the seminiferous tubules from January to April and November to December,and mainly in spermatogonia,primary spermatocytes,secondary spermatocytes and circles from May to October,and there is a seasonal difference in the rate of positive cells;The p21 protein and p53 protein had no immunopositive signals on spermatogenic cells at all levels in the tubules of tree shrews in each month.The results of in situ hybridization showed that in the artificially domesticated tree shrew testis tissues from January to December,the positive signal of MCM7 gene hybridization was mainly detected in spermatogonia and primary spermatocytes in the tubule;The positive signal of Bcl-2 gene hybridization was detected in the spermatogonia and primary spermatocytes of the seminal tubules of the tree shrew testis from January to December;The positive hybridization signal of Bax gene was detected in spermatogenic cells at all levels in February,in spermatogonia and primary spermatocytes in January,March,September and October,and in spermatogenic cells at all levels from April to August None detected,detected in spermatogonia,primary spermatocytes,secondary spermatocytes and round spermatids from November to December;The positive signal of p21 gene hybridization was detected in spermatogenic cells at all levels in the seminal tubules of the tree shrew testes in January and February,and the positive hybridization signal was detected in the spermatozoa in the seminal tubules from March to May.and positive hybridization signal was detected in spermatogonia and primary spermatocytes from October to December;The positive signal of p53 gene hybridization was detected in the seminiferous tubules of the tree shrew testicles from February to April and August to November,and no positive was detected in the spermatogenic cells of the seminiferous tubules from May to July For hybridization signals,weaker positive hybridization signals were detected in spermatogenic cells at all levels of Seminiferous tubules in December and January.Conclusions:1.During spermatogenesis of artificially domesticated tree shrews,there are seasonal differences in the microstructure of the testicular seminiferous tubules;2.During spermatogenesis of artificially domesticated tree shrews,there are seasonal differences in the submicroscopic structure of the seminiferous tubules of the testes;3.MCM7,Bcl-2 and Bax are involved in regulating the spermatogenesis of artificially domesticated tree shrew testes,and there are seasonal differences in their expression in spermatogenic cells;4.The p21 and p53 may not be involved in the regulation of spermatogenesis in artificially domesticated tree shrews at the protein level.