• Journal of Animal Science and Technology
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• 제55권4호
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• pp.249-255
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• 2013

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.

• Journal of Animal Science and Technology
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• 제63권2호
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• pp.262-271
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• 2021

Communication among epididymal epithelial cells creates the best luminal condition where spermatozoa mature, transport and are stored. Vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) have been used as signal indicators for clear and basal cells of the epididymal epithelium, respectively, in mice, rats, bats, and pigs; however, these two markers have not yet been described in the epididymis of bulls. Here, we examined the presence and distribution of the B1 subunit of V-ATPase (B1-VATPase) and KRT5 in the distinct regions of adult bovine epididymides, specifically, the caput, corpus, and cauda. Immunofluorescence staining and confocal microscopy showed that narrow shaped-clear cells were placed in the caput and corpus regions of the bovine epididymis; however, they were absent in the cauda epididymis. In addition, B1-VATPase was highly expressed in the cauda spermatozoa; however, it was rarely detected in the caput spermatozoa. On the other hand, KRT5-positive cells, basal cells, were maintained beneath the basal lamina and they had the traditional form with a dome-shaped morphology from the caput to cauda region of the bovine epididymis. The co-expression of B1-VATPase and KRT5 was confined to basal cells placed in the basal region of the epithelium. In summary, 1) clear cells were present with region-specific localization, 2) B1-VATPase was present in the corpus and cauda spermatozoa but absent in the caput, 3) co-expressed cells with B1-VATPase and KRT5 were present in the adult bovine epididymis, and 4) B1-VATPase was not a specific marker for clear cells in the bovine epididymis. Therefore, the perfect epididymal luminal condition created by the specific expression and localization patterns of B1-VATPase might be necessary to obtain fertilizing capacity of spermatozoa in the bovine epididymis.

• Asian-Australasian Journal of Animal Sciences
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• 제5권1호
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• pp.165-171
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• 1992

The developmental process of the epididymis was investigated in Meishan boars from 1 to 364 days of age. Epididymal weight increased rapidly between 45 and 150-180 days of age. The diameter and epithelial area of the epididymal ducts greatly increased up to 105-120 days of age. At 1 day of age, the central and distal cauda already had a pseudostratified epithelium surrounded by smooth muscle. At 60-75 days of age, the central and distal caput, corpus, and proximal cauda revealed a well-developed structure of the epithelium. The proximal caput showed a tall, irregular and vacuolar epithelium at 105-120 days of age. PAS-positive contents in the lumen of the caput, corpus and cauda epididymides were first detected at 60-75, 45-60 and 1-30 days of age, respectively. Moreover, in the central and distal caput, PAS-positive granules appeared at 60-75 days of age, and increased until 105-120 days. These results suggest that the epididymis develops completely by approximately 120 days of age, though its weight increases rapidly up to 150-180 days. Thus, it appears that development of the epididymis occurs at an earlier age in Meishan boars than in European and American breeds.

• Applied Microscopy
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• 제22권1호
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• pp.89-102
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• 1992

This research was undertaken to determine the effects of the anticancer and immunosuppressive drug cyclophosphamide (CP) on the epididymal caput of the male rat in terms of ultrastructural alteration and protein analysis by SDS-PAGE at different groups; control group, 1 week group, 3 weeks group, 5 weeks group were treated with saline (control group) or CP at doses of 20 mg/kg/week, 1 time a week, respectively. In the cytoplasm of the principal cells on the epididymis, the mitochondrial outer and inner membranes were significantly swollen or disrupted. The lumens of rough endoplasmic reticulum (rER) were also swollen, and the number of Golgi vesicles were increased, respectively. It is suggested that treatment with CP alters the specific cell organelles in the epididymis. CP caused changes in protein concentrations in caput of epididymis after CP treatment. Total proteins of 32 to 37 species such as lactate dehydrogenase, carnitine acetyltransferase and succinate dehydrogenase were expressed in the caput fluid. Then the more CP was increased, the more concentration of proteins caused to decrease, synthesize or increase in epididymis. In contrast to the control group, in particular carnitine acetyltransferase and the other 9 proteins in the caput fluid were decreased or disappeared, respectively, whereas lactate dehydrogenase and the other 5 proteins in the caput fluid were increased or synthesized, respectively. The other proteins are not showed distinctive difference. These alterations could be direct mediated by toxic effects of the drug on the epithelium or be secondary to changes in the spermatozoa as a result of the CP treatment.

• 한국발생생물학회지:발생과생식
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• 제21권3호
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• pp.317-325
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• 2017

Direct communication between neighboring cells through connexin (Cx)-based gap junction is a crucial biological manner to regulate functions of a tissue consisting of multi-cell types. The present research evaluated expressional changes of Cx isoforms in the caput epididymis of adult rat exposed to estradiol benzoate (EB) or flutamide (Flu) at the early postnatal age. A single subcutaneous administration of EB at a low-dose [$0.015{\mu}g/kg$ body weight (BW)] or a high-dose ($1.5{\mu}g/kg\;BW$) or Flu at a low-dose ($500{\mu}g/kg\;BW$) or a high-dose (5 mg/kg BW) was performed to an animal at 1 week of age. Quantitative real-time PCR analysis was employed to determine expressional changes of Cx isoforms. The transcript levels of Cxs30.3 and 37 were decreased by a low-dose EB treatment, while decreases of Cxs31, 31.1, 32, 40, and 45 transcript levels were observed with a low-dose EB treatment. The treatment of a high-dose EB resulted in expressional reduction of Cxs30.3, 31, 31.1, 37, 40, 43, and 45. The Flu treatment at a low dose caused increases of Cxs26, 37, and 40 transcript levels but decreases of Cxs31.1, 43, and 45 transcript levels. Increases of Cxs30.3, 31, 37, and 40 mRNA amounts were induced by a high-dose Flu treatment. However, exposure to a high-dose Flu produced expressional decreases of Cxs31.1, 32, and 43 in the adult caput epididymis. These observations suggest that exposure to EB or Flu at the neonatal period could lead to aberrant expression of Cx isoforms in the adult caput epididymis.

• Asian-Australasian Journal of Animal Sciences
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• 제29권12호
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• pp.1702-1709
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• 2016

The reproductive function of G-protein subunit Galphaq (GNAQ), a member of the G protein alpha subunit family, has been extensively studied in humans and rats. However, no data is available on its status in ruminants. The objectives of this study were to evaluate the expression pattern of the GNAQ in the testis and epididymis of sheep by polymerase chain reaction (PCR). The mRNA expression levels were detected by real-time fluorescent quantitative PCR, and cellular localization of GNAQ in the testis and epididymis was examined by immunohistochemistry. Additionally, GNAQ protein was qualitatively evaluated via western blot, with the results indicating that similarities between GNAQ mRNA levels from sheep was highly conserved with those observed in Bos taurus and Sus scrofa. Our results also indicated that GNAQ exists in the caput and cauda epididymis of sheep, while GNAQ in the testis and epididymis was localized to Leydig cells, spermatogonial stem cells, spermatocytes, Sertoli cells, spermatid, principal cells, and epididymis interstitial cells. The concentrations of GNAQ mRNA and protein in the caput and cauda epididymis were significantly greater than those observed in the corpus epididymis (p<0.01) and testis (p<0.05). Our results indicated that GNAQ exists at high concentrations in the caput and cauda epididymis of sheep, suggesting that GNAQ may play an important role in gonad development and sperm maturation.

• Advances in Traditional Medicine
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• 제8권1호
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• pp.67-72
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• 2008

Adult male albino rats were treated with 0.5 mg and 0.75 mg morphine/100 g body weight intraperitoneally for 30 days. All the animals were autopsied on $31^{st}$ day. Epididymis and vas deferens were dissected out, weighed and processed for histological and biochemical studies. Morphine has caused a reduction in the weight of epididymis and vas deferens in both the doses of drug treated groups. The total cholesterol content is increased while protein, DNA and RNA contents and epididymal sperm counts are decreased. The acid phosphatase content is decreased 10.12 $\pm$ 0.11 in caput, 9.26 $\pm$ 0.30 in cauda of epididymis and in vas deferens 8.14 $\pm$ 0.15 in 0.5 mg treated groups and in 0.75 mg treated rats shows 9.52 $\pm$ 0.27 in caput, 9.14 $\pm$ 0.18 in cauda of epididymis and in vas deferens 7.84 $\pm$ 0.11is decreased, whereas alkaline phosphatase is increased. The surface epithelial cell height of these ducts is reduced and secretory activity is inhibited with the disruption of epithelial cell projections. The gravimetric and histometric changes of epididymis and vas deferens may be due to non-availability of androgens in morphine treated rats.

• 한국동물학회지
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• 제34권2호
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• pp.159-172
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• 1991

흰쥐 부정소 정자의 성숙 전후에 일어나는 변화를 몇가지 효소를 중심으로 관찰하였고 그 성숙과정중에 일어나는 상피세포, 내강 및 정자 사이의 상호관계를 알아보기 위하여 실험군 별로 몇가지 효소의 활성도와 탄수화물 잔기의 함량을 측정하였으며, 전기영동을 이용하여 각 군의 차이를 관찰하고 이에 대한 웅성호르몬의 관련성을 알아보았다. 1. 부정소 두 정자와 부정소미 정자에서 활성도 측정시 lactate dehydrogenase, glucose-6-phosphatase 및 Na+ -K+ -ATPase의 경우는 유의한 차이가 없었으며, $Mg^2$+-ATPase의 경우만이 부정소미 정자가 부정소두 정자보다 유의성 있게 높은 활성도를 가지는 것으로 나타났다. 또한 부정소두와 부정소미 상피세포 , 내강 및 정자의 세군으로 나누어 각각 탄수화물 잔기를 정량하였을 때 hexosamine은 상피세포에서, sialic acid는 상피세포와 내강액에서 부정소미의 경우가 더 높은 함량이 존재하였으며, 내강액과 정자의 crude membrance fraction을 SDS-PAGE 했을 때 분자량이 33-37 KD 사이에 존재하던 band가 부정소미 내강액과 부정소미 정자의 crude membrance fraction에서 관찰되었으므로 흰쥐에서 정자의 성숙과정과 관련된 부정소내의 여러 변화를 비교하는 자료가 될 수 있었다. 2. 부정소 내강액에서 $\beta$ -glucuronidase와 $\beta$ -glucosidase의 활성도 및 웅성호르몬에 대한 의존성을 측정하였을 때 거세 후 5일째부터 이 두 효소의 활성도가 모두 유의하게 감소하기 시작하였고, tentosterone을 투여하였을 때는 $\beta$ -glucuronidase는 투여 5일, $\beta$ -glucosidase는 투여 10일 후부터 유의하게 증가하였으며 웅성호르몬에 대한 내강액의 의존성을 알아보기 위하여 SDS-PAGE하였을 때 tentosterone투여군의 부정소두에서 분자량이 약 21 KD에 해당하는 band를 새로이 관찰하였다.

• 한국동물생명공학회지
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• 제34권3호
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• pp.190-196
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• 2019

Tight junctions are constituents of the blood-epididymis barrier that play roles in regulating the unidirectional transcellular transport of ions, water, and solutes to maintain optimal conditions for sperm maturation and storage. Claudin 1 (Cldn1) and 4 (Cldn4) are known as tight junction proteins and are expressed in the basolateral membranes as well as tight junctions in the epididymis of rodents. Here, we examined the expression and localization of Cldn1 and 4 to determine the function of these proteins in the pig epididymis. Cldn1 was highly expressed in the basolateral membrane of epithelial cells in the caput and corpus regions of the epididymis. In the cauda region, however, Cldn1 labeling was significantly decreased in the basolateral membrane of epithelial cells. In contrast, labeling indicated that Cldn4 was expressed in the basolateral membrane in the cauda region of the epididymis and was present at punctate reactive sites in the caput and corpus regions. However, in no region of the epididymis did we detect colocalization of Cldn1 and 4 with labeled ZO-1, the distribution of which is restricted to the tight junctions. Our results indicate that Cldn1 and 4 were region-specifically expressed in the pig epididymis but not present in the tight junctions of epididymal epithelium. In addition, reciprocal regulation in specific regions of the epididymis between Cldn1 and 4 may play an important role in generating an optimal luminal environment for sperm maturation and storage in the pig epididymis.

• Asian-Australasian Journal of Animal Sciences
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• 제33권7호
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• pp.1077-1086
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• 2020

Objective: We examined the localization and expression of H⁺ pumping vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) in the epididymis of pigs, expressed in clear and basal cells, respectively, during postnatal development. Methods: Epididymides were obtained from pigs at 1, 7, 21, 60, 120, and 180 days of age; we observed the localization and expression patterns of V-ATPase and KRT5 in the different regions of these organs, namely, the caput, corpus, and cauda. The differentiation of epididymal epithelial cells was determined by immunofluorescence labeling using cell-type-specific markers and observed using confocal microscopy. Results: At postnatal day 5 (PND5), the localization of clear cells commenced migration from the cauda toward the caput. Although at PND120, goblet-shaped clear cells were detected along the entire length of the epididymis, those labeled for V-ATPase had disappeared from the corpus to cauda and were maintained only in the caput epididymis in adult pigs. In contrast, whereas basal cells labeled for KRT5 were only present in the vas deferens at birth, they were detected in all regions of the epididymis at PND60. These cells were localized at the base of the epithelium; however, no basal cells characterized by luminally extending cell projections were observed in any of the adult epididymides examined. Conclusion: The differentiation of clear and basal cells progressively initiates in a retrograde manner from the cauda to the caput epididymis. The cell-type-specific distribution and localization of the epithelial cells play important roles in establishing a unique luminal environment for sperm maturation and storage in the pig epididymis.