Expression of connexin 43 protein in testes, epididymides and prostates of stallions

Hejmej, Anna; Kotula–Balak, Małgorzata; Sadowska, J; Bilińska, Barbara

doi:10.2746/042516407x169393

Cited by 48 publications

(37 citation statements)

References 31 publications

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“…Of importance, in bison testes with impaired spermatogenesis, the Cx43 signal was significantly reduced or even absent. The results are consistent with earlier observations on pathological testes of mouse, horse, and human (Steger et al, '99;Batias et al, 2000;Hejmej et al, 2007;Kotula-Balak et al, 2007) in which a close correlation between the level of Cx43 expression and severity of seminiferous tubules degeneration …”

Section: Discussionsupporting

confidence: 82%

“…Our finding that the Cx43 localizes between Leydig cell plasma membranes is in agreement with previously published observations on rat, guinea pig, and equine testes (Risley et al, '92;Pelletier, '95;You et al, 2000;Hejmej et al, 2007). Of all the connexins tested so far in the testis, Cx43 is the only connexin detected in Leydig cells (reviewed by .…”

Section: Discussionsupporting

confidence: 82%

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Age‐dependent pattern of connexin43 expression in testes of European bison (Bison bonasus, L.)

et al. 2009

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Morphology and the expression of connexin43 (Cx43) was investigated in testes of immature, prepubertal, and adult European bison bulls by means of routine histology, immunohistochemistry, and Western blotting, respectively. Testes were collected from culled animals living in Bialowieza and Borecka Forests, Poland. Histological examination of testicular tissue of immature and prepubertal males revealed normal structure, whereas of adult individuals either normal testicular structure with advanced spermatogenesis or varying degrees of tubule and interstitial tissue abnormality were seen. Immunohistochemical studies revealed Cx43 signal mostly at Leydig cell membrane appositions. In testes of immature males heterogeneous staining was observed; its intensity markedly increased in prepubertal males reaching almost two times more intense staining in adults. Strong Cx43 signal between Leydig cells was also observed in testes with structural alterations; however, sporadically it was of weak linear pattern. In the tubules of the latter, the intensity of Cx43 staining was weak to moderate or it was even absent. Immunohistochemical qualitative analysis was confirmed by quantitative image analysis in which the staining intensity was expressed as relative optical density of diaminobenzidine deposits. Data from Western blot analyses confirmed the results obtained by immunohistochemistry; immunodetectable Cx43 protein as a band of 43 kDa was detected in all testes samples. Overall, the increase in Cx43 level in testes along the bison postnatal development may be capable of a better exchange of metabolites and coordinating Leydig cell activity during maturation. A relationship between homozygosity occurring in European bison and altered intercellular communication is suggested.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionsupporting

confidence: 82%

Age‐dependent pattern of connexin43 expression in testes of European bison (Bison bonasus, L.)

et al. 2009

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“…In the stallion, the cryptorchid results in a significant decrease of Cx43 expression (Hejmej et al, 2007). Recently, Dubé et al (2012) have reported the regulation of Cx43 expression by epidermal growth factor in the human epididymis.…”

Section: Direct Intercellular Communication Throughmentioning

confidence: 99%

The Expression Patterns of Connexin Isoforms in the Rat Caput Epididymis During Postnatal Development

Han¹,

Lee²

2013

Journal of Animal Science and Technology

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Intercellular interactions are important for the proper development and regulation of tissue function. This is especially necessary in the epididymis, a part of male reproductive tract where sperm become mature and acquire their fertilizing capacity. The caput region of the epididymis consists of several types of cells, including principal, basal, and apical cells. Direct intercellular communication is thus required to precisely regulate the functions of the caput epididymis. In this regard, connexin (Cx) is a molecule that forms channels, which allow the direct exchange of small molecules between cells, enabling intercellular communication. In this study, the expression of Cx isoforms in the caput epididymis at different postnatal ages was determined by using quantitative real-time polymerase chain reaction analysis. Nine of 13 Cx isoforms were detected. The transcript levels of Cx30.3, 31, 31.1, 32, and 40 were highest at 45 days of age, while the expression of Cx43 and 45 gradually decreased with age. A substantial fluctuation of Cx26 expression was detected, with significant decreases before and during puberty, followed by a transient increase at adult-hood and rapid decreases at an old age. A significant increase in Cx37 transcript was observed at 25 days of age, followed by gradual decreases at adult and old ages. These results indicate the significant differential expression of various Cx isoforms in the caput epididymis during postnatal development. It further suggests that the functional regulation and developmental maturation of the caput epididymis are highly related to the postnatal age-related differential expression of Cx isoforms.

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“…Testes were washed twice with ice-cold saline, and proteins were then extracted in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Thermo Scientific, Inc., Rockford, IL, USA) supplemented with protease and phosphatase inhibitor cocktails (Sigma-Aldrich) as described previously (Hejmej et al, 2007). Aliquots were assayed for protein using the D C protein assay with bovine serum albumin as a standard (Bio-Rad Laboratories).…”

Section: Western Blot Analysismentioning

confidence: 99%

Flutamide alters the distribution of c-Src and affects the N-cadherin-β-catenin complex in the seminiferous epithelium of adult rat

et al. 2015

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Flutamide alters the distribution of c-Src and affects the N-cadherinb-catenin complex in the seminiferous epithelium of adult rat SUMMARYThis study was undertaken to explore interactions between c-Src kinase and the N-cadherin-b-catenin complex in seminiferous tubules of flutamide-treated rats. An anti-androgen flutamide (50 mg/kg bw) was injected daily into adult rats from postnatal days 82 to 88. Testes from 90-day-old control and flutamide-treated rats were used for experiments. Flutamide did not affect testis morphology, but impaired connexin43 immunoexpression between Sertoli cells at the blood-testis barrier (BTB) region, indicating the BTB as a sensitive target for flutamide. Real-time RT-PCR and western blot analyses revealed upregulation of N-cadherin at the mRNA and protein level after flutamide exposure (p < 0.05), whereas no changes in b-catenin and c-Src expression were observed. Notably, membranous b-catenin immunolocalization indicated its involvement in the cell adhesion complex rather than its contribution to the Wnt signaling pathway. As we used an exposure regime which avoided germ cell loss, it is likely that changes in the N-cadherin-b-catenin complex are a primary effect of androgen signaling disruption by flutamide. Immunohistochemistry revealed a diffusion of N-cadherin and b-catenin signals away from the BTB with concomitant disruption of c-Src staining pattern. As detected by immunofluorescence and coimmunoprecipitation, flutamide promoted disassembly of the N-cadherin-ß-catenin complex, induced N-cadherin to dissociate from c-Src at the BTB site, and altered interactions between the cell junction proteins and/or c-Src. Equally important, increased levels of p-N-cadherin-Tyr860 and p-b-catenin-Tyr654 (p < 0.05) pointed to a mechanism related to adhesion complex disassembly and suggested a potential role of c-Src in the control of the protein-protein dynamics. Overall, for the first time we have shown that flutamide alters the distribution of c-Src and affects N-cadherin-b-catenin interactions at the BTB. Understanding mechanism(s) by which anti-androgens can affect intercellular adhesion within the testis is relevant for predicting and preventing reproductive disorders affecting male fertility.

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Expression of connexin 43 protein in testes, epididymides and prostates of stallions

Abstract: Detection of Cx43 expression in equine testicular, epididymal, and prostatic cells is important for a better understanding of the role of intercellular membrane channels in direct cell communication within the reproductive tract of stallions.

Cited by 48 publications

References 31 publications

Age‐dependent pattern of connexin43 expression in testes of European bison (Bison bonasus, L.)

Age‐dependent pattern of connexin43 expression in testes of European bison (Bison bonasus, L.)

The Expression Patterns of Connexin Isoforms in the Rat Caput Epididymis During Postnatal Development

Flutamide alters the distribution of c-Src and affects the N-cadherin-β-catenin complex in the seminiferous epithelium of adult rat

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