Abstract
Neuroendocrine substances play essential roles in regulating the normal physiological functions of testicles. The purpose of this study is to explore the localization and effects of four neuroendocrine markers (NSE, SP, NFH and DβH) in normal and cryptorchid testes of Bactrian camels using western blotting, transmission electron microscopy, immunohistochemistry, and immunofluorescence methods. The results showed that cryptorchidism caused a reduction in layers of spermatogenic epithelium and decreased glycogen positivity in the basement membrane. The ultrastructure revealed that macrophages were always found around the Leydig cells, crowded with swelling mitochondria in cryptorchidism. Expression of NSE in the Leydig cells of cryptorchidism was significantly weakened compared to that in the normal group(p<0.01). We found that SP was always distributed along the nerve fibers in normal testes and was expressed in the Leydig cells of cryptorchidism. However, expression of NFH in the cryptorchidic tissue was strongly positive in the spermatogenic epithelium, with limited expression in Leydig cells and no expression in peritubular myoid cells. Therefore, the expression of DβH in the Sertoli cells was comparatively strong in both the normal and cryptorchidism groups. NFH and DβH expression was significantly increased in the cryptorchidism group compared with the normal group (p<0.01). These findings indicated that the underdeveloped seminiferous epithelium and pathological changes in cryptorchid tissue in Bactrian camels were potentially related to a disorder in glycoprotein metabolism. Our results suggest that NSE and SP could help judge the pathological changes of cryptorchidism. The present study provides the first evidence at the protein level for the existence of NFH and DβH in Sertoli and Leydig cells in Bactrian camel cryptorchidism and provides a more in-depth understanding of neuroendocrine regulation is crucial for animal cryptorchidism.
Highlights
Gametogenesis in mammals is a very complex process in which paracrine and autocrine mechanisms in addition to the hypothalamic-pituitary-testicular endocrine axis influence are intimately involved in complicated feedback loops (Plant, 2015; Khanehzad et al, 2021)
We detected glycogen by PAS staining and distinct purple-red glycogen-positive bands in the lamina propria and interstitial capillary walls were observed (Fig. b); Acid mucins was clearly visible in the interstitial capillary wall and lamina propria as demonstrated by AB staining (Fig. c); Examination of the H&E images revealed that cryptorchidism causes a reduction in layers of spermatogenic epithelium, the Leydig cells lacked distinct structural characteristics, and the nucleus was round, elliptical, or irregular, with relatively large cell bodies (Fig. d); We examined whether r a decrease in glycoprotein metabolism caused the reduction in seminiferous tubule cross-section
The sulfated glycoproteins protect the normal activity of sperm (Liu, 2016).Our studies revealed that the cryptorchidism decreased the diameter of the Leydig cell (p< 0.05) and induced glucose metabolism disorders in the surrounding connective tissue, which may contribute to the abnormal distribution of local neurohormones in testis
Summary
Gametogenesis in mammals is a very complex process in which paracrine and autocrine mechanisms in addition to the hypothalamic-pituitary-testicular endocrine axis influence are intimately involved in complicated feedback loops (Plant, 2015; Khanehzad et al, 2021). Expression of NSE,SP,NFH and DβH in Cryptorchidism distribution and localization of PGP9.5,neuropeptide Y, neuron-specific enolase (NSE), substance P(SP), dopamine-β-hydroxylase (DβH), and tyrosine hydroxylase-related peptides in the testicles of mammals, such as cats (Suburo et al, 2002), donkeys (Wrobel and Moustafa, 2000), ruminants (Maddocks et al, 1995), dromedaries (Saleh et al, 2002) and humans (Gong et al, 2009) were studied by immunohistochemistry. Results showed that such neuroendocrine substances play essential roles in regulating the normal physiological functions of the testicles. These results are necessary to clarify the capacity of Leydig cells to synthesize these substances and establish their functional significance in reproduction
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