Expression And Distribution Of Sex Hormone Receptor And Related Factor On Hypothalamic-pituitary-testicular Axis In Bactrian Camel Cryptorchidism

Bactrian camel is an important dominant animal species in China,but its reproductive rate is low.Cryptorchidism,as one of the reproductive disorders of Camelidae,has a great impact on its reproductive function.As the site of hormone secretion and sperm production,testis plays an important role in the reproduction of Bactrian camel.Cryptorchidism was due to not normal testis decreased,the local temperature,gonad axis hormones,the serious influence of mammalian sperm and semen quality.The aim of this study is to investigate the localization and gene and protein expression distribution of AR,ERβ,LHR,and LC λ in the hypothalamus,pituitary gland,and testis of normal and cryptorchidism Bactrian camel,and to explore the mechanism of cryptorchidism,and to provide reference for the testicular pathology and reproductive physiology of Bactrian camel.In this experiment,a variety of special staining methods were used to observe the change characteristics of protein and glycogen in normal and cryptorchidism Bactrian camel testis.Meanwhile,routine tissue section staining H&E method was used to observe the changes of hypothalamus,pituitary and testis tissue morphological characteristics of normal and cryptorchidism Bactrian camel.Secondly,immunohistochemistry(IHC)was used to identify the distribution and expression of sex hormone receptors and inflammatory factors in the hypothalamus,pituitary gland and testis of normal and cryptorchidism Bactrian camels.Then,quantitative reverse transcription PCR(RT-q PCR)and Western blot(WB)were used to compare the expression levels of gene and protein.The results show that:1.The expression of protein and glycogen in cryptorchidism of Bactrian camels was abnormal.Compared with normal testes,the distribution of total protein in cryptorchidism was weakened,the content of basic protein was decreased,and the distribution of acid and neutral mucin was decreased,suggesting that the microenvironment of spermatogenesis and the distribution of glycogen and other nutrients were abnormal during cryptorchidism.2.Compared with normal Bactrian camel,there was no significant difference in the morphology of hypothalamus and pituitary in cryptorchidism Bactrian camel,but there was a significant change in the morphology of testis,which showed a decrease in the lumen diameter of cryptorchidism seminiferous tubules,a decrease in the number of spermatogenic epithelial cell layers,a delay in the development of testicular Sertoli cells and various types of spermatogenic cells,and abnormal morphology of cryptorchidism.3.The results of IHC showed that AR,ERβ and LHR were mainly expressed in nerve cells and granulosa cells in normal and cryptorchidism Bactrian camel hypothalamus tissue,suggesting that AR,ERβ and LHR may play a role in brain physiology and reproductive behavior.LC λ was only positively expressed in granule cells,suggesting that hypothalamic granule cells may play a role in mediating the development of inflammation.Normal and cryptorchidism Bactrian camel pituitary tissues,AR mainly locate in eosinophilic cells;ERβ was positively expressed in eosinophilic cells,basophilic cells and chromophobes.LHR was mainly expressed in eosinophilic cells and basophilic cells.LC λ was positively expressed only in chromophobe cells.In normal and cryptorchidism testis,AR was strongly expressed in Sertoli cells,Leydig cells and spermatogonia.ERβ was expressed in all spermatogenic cells,and was strongly positive in Leydig cells,primary spermatocyte and peritubular myoid cells.LHR was expressed in Leydig cells and other spermatogenic cells,but not in peritubular myoid cells.LC λ was expressed only in Sertoli cells in normal testis,but in all kinds of cells in cryptorchidism.4.RT-q PCR and Western-blot results showed that compared with the normal Bactrian camel hypothalamic-pituitary-testis axis,the expression levels of AR gene and protein in the cryptorchidism Bactrian camel were significantly down-regulated,while the expression of LHR and LC λ was significantly up-regulated.There was no significant difference in the expression of ERβ.In conclusion,the hypothalamus and pituitary gland of cryptorchidism Bactrian camel have little difference in histomorphology compared with normal Bactrian camel,but the secretion and tissue morphology of cryptorchidism glycogen and related proteins are abnormal,affecting reproductive function.In the hypothalamic-pituitarytestis axis of cryptorchidism,the secretion of hypothalamic and pituitary AR and LHR is abnormal,and the expression of AR in testicular Leydig cells is significantly different.LHR is closely related to the development of Sertoli cells in the testis,LC λ is closely related to the oxidative stress and inflammation in the testis during the occurrence of cryptorchidism,while ERβ has a weak effect on the testis.