Steroid hormonal regulation on Foxi1 expression in mouse epididymis

Jae Eun Lee,Il Kyoo Koh,Myung Chan Gye 한국발생생물학회 2011 한국발생생물학회 학술발표대회 Vol.30 No.-

Foxi1, a forkhead family of transcription factor, in narrow and clear cells in epididymis is required for male fertility through regulating transcription of vacuolar H+-ATPase. To understand the regulation of Foxi1 gene activation in epididymis, the effects of steroids and their receptor antagonists and testicular factors on the expression of Foxi1 in epididymal segments were examined in mouse. Epididymis were sampled from adult mice following injections of ICI 182,780 (5mg/head, 2 times for 15 days), dexamethasone (DEX, 0.1,1,10ug/kg/day for 5 days) or oral administration of flutamide (FLM, 100mg/kg/day for 10 days). Otherwise, adult mice were orchidectomized (ORX), rested for 2 weeks, and received testosterone propionate(TP, 3mg/kg/day) for 7 days. In addition, adult male mice were subjected to efferent duct ligation (EDL) and epididymis was collected after 15 days. To study estrogen regulation of Foxi1 gene activation via estrogen receptor α (ESR1), Foxi1 expression was examined in ESR1 knock-out mice epididymis. Expression and subcellular localization of Foxi1 was analyzed by realtime RT-PCR and immunohistochemistry. To search transcription factor binding in the mouse Foxi1 gene promoter, in silico analysis was performed using TESS, TFSEARCH, and Gene-Regulation. ICI 182,780 significantly decreased Foxi1 mRNA levels in caput and corpus but increased in cauda epididymis. Foxi1 mRNA levels in caput epididymis of ESR1 KO mice were significantly lower than those of WT mice, but no significantly changed in corpus and cauda epididymis. Taken together, estrogen differentially regulates Foxi1 gene expression in epididymis. In ORX mice, Foxi1 mRNA levels were significantly increased in epididymis, and which was abrogated by TP. Though FLM did not significantly alter the Foxi1 mRNA levels, androgen may affect Foxi1 gene expression in epididymis. DEX significantly decreased Foxi1 mRNA levels in caput and corpus epididymis at 0.1ug/kg/day and in cauda epididymis at 1ug/kg/day, suggesting that glucocorticoid may negatively regulate Foxi1 gene expression. No significant change in Foxi1 mRNA levels was found after EDL. Foxi1 immunoreactivity was found in the nuclei of narrow cells of caput epididymis including initial segment and clear cells of corpus and cauda epididymis. Of note, in ORX mice, Foxi1-positive narrow cells and clear cells were increased, and which was abrogated by TP. In silico analysis revealed the presence of putative binding sequences for ESR1, AR, and GR in the 5’ upstream region from the Foxi1 promoter. In conclusion, the expression of Foxi1 in narrow cells in caput epididymis might be positively regulated by estrogen via ESR1, which was different from estrogen–ESR signaling in clear cells in corpus and cauda epdididymis. Androgen and glucocorticoid may negatively regulate expression of Foxi1 in all epdididymial segments.

Effect of an Anabolic Steroid, Nandrolone Decanoate, on Aquaporin 1 and 9 Gene Expression in the Rat Epididymis

Hee-Jung Seo,Hyo-Jin Kang,Inho Choi,Yong-Pil Cheon,Ki-Ho Lee 한국동물생명공학회(구 한국동물번식학회) 2009 Reproductive & developmental biology Vol.33 No.1

The epididymis in the male reproductive tract is the site where spermatozoa produced from the testis become mature. The epididymis is divided into 4 different segments, initial segment and caput, corpus, and caudal epididymis, depending upon functional and morphological features. Aquaporins (Aqps) are water channel molecules, which are present in the epididymis and play a major role in removal of epididymal water, resulting in creation of microenvironment for sperm maturation and concentration of sperms. Nandrolone decanoate (ND) is a synthetic anabolic-androgenic steroid, which is used to treat clinical diseases and improve physical ability and appearance. Even though it is well determined that the ND causes the male infertility by affecting the testis, little is known the effect of the ND on the epididymis. The present study was focused to examine the effect of ND at different treatment doses and periods on expression of Aqp1 and Aqp9 genes in the epididymis of pubertal rats. Results showed that mRNA expression of Aqp1 and Aqp9 genes among the parts of the epididymis was differentially regulated by ND treatment doses. In addition, treatment periods of ND caused differential expression of Aqp1 and Aqp9 mRNAs among segments of the epididymis. Therefore, it is believed that male infertility induced by ND could be resulted not only from malfunction of the testis but also from aberrant gene expression of Aqp1 and Aqp9 in the epididymis.

돼지 정소상체에서 ZO-1, Claudin 1 및 Claudin 4의 발현 양상

박윤재,김봉기 사단법인 한국동물생명공학회 2019 한국동물생명공학회지 Vol.34 No.3

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.

Effect of an Anabolic Steroid, Nandrolone Decanoate, on Aquaporin 1 and 9 Gene Expression in the Rat Epididymis

Seo, Hee-Jung,Kang, Hyo-Jin,Choi, In-Ho,Cheon, Yong-Pil,Lee, Ki-Ho The Korean Society of Animal Reproduction 2009 Reproductive & developmental biology Vol.33 No.1

The epididymis in the male reproductive tract is the site where spermatozoa produced from the testis become mature. The epididymis is divided into 4 different segments, initial segment and caput, corpus, and caudal epididymis, depending upon functional and morphological features. Aquaporins (Aqps) are water channel molecules, which are present in the epididymis and play a major role in removal of epididymal water, resulting in creation of microenvironment for sperm maturation and concentration of sperms. Nandrolone decanoate (ND) is a synthetic anabolic-androgenic steroid, which is used to treat clinical diseases and improve physical ability and appearance. Even though it is well determined that the ND causes the male infertility by affecting the testis, little is known the effect of the ND on the epididymis. The present study was focused to examine the effect of ND at different treatment doses and periods on expression of Aqp1 and Aqp9 genes in the epididymis of pubertal rats. Results showed that mRNA expression of Aqp1 and Aqp9 genes among the parts of the epididymis was differentially regulated by ND treatment doses. In addition, treatment periods of ND caused differential expression of Aqp1 and Aqp9 mRNAs among segments of the epididymis. Therefore, it is believed that male infertility induced by ND could be resulted not only from malfunction of the testis but also from aberrant gene expression of Aqp1 and Aqp9 in the epididymis.

돼지 정소상체에서 ZO-1, Claudin 1 및 Claudin 4의 발현 양상

박윤재,김봉기 한국동물생명공학회(구 한국동물번식학회) 2019 Journal of Animal Reproduction and Biotechnology Vol.34 No.3

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.

Evaluation of sperm number, motility, plasma membrane integrity on different regions of epididymis in Hanwoo bull

Sung-Sik Kang,Ui-Hyung Kim,Myung-Suk Lee,Seok-Dong Lee,Sun-Sik Jang,Ki-Yong Chung,Hyun-Joo Kim,Sang-Rae Cho 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

In this study, we examined number, motility and plasma membrane integrity of spermatozoa from six regions of epididymis in bull. Six testicles with epididymides were castrated from six bulls (mean±standard error, age of days = 441.3±9.6, body weight (kg) = 367±8.4, scrotal circumference (cm) = 30.7±0.4) at Hanwoo Research Institute, NIAS and transported to laboratory within 1 hour. Testicular weight, length, width and circumference were recorded. Epididymis in each bull was randomly used for recovery of spermatozoa. Epididymis was divided into six regions: efferent duct (ED), caput, corpus, proximal cauda (Pcauda), distal cauda (Dcauda) and vas deferens (VD). In experiment 1, we examined sperm number of each region of epididymis. Each region of epididymis contained different number of spermatozoa: ED (37.8±15.7 × 106cells/ml, 8.2%), caput (93.6±18.8 × 106cells/ml, 20.2%), corpus (33.0±8.5 × 106cells/ml, 7.1%), Pcauda (104.2±23.5 × 106cells/ml, 22.5%), Dcauda (180.5±32.5 × 106cells/ml, 39.0%) and VD (14.0±5.0 × 106cells/ml, 3.0%). In experiment 2, sperm motility of each epididymal region was examined by computer assisted sperm analysis (SCA, MicroOptic) system. Sperm motility was divided into 4 groups (fast progressive, slow progressive, non-progressive and immotile) based on WHO guideline. Percentages of fast progressive of Pcauda and Dcauda (11.0±2.3 and 15.4±3.6%) were significantly higher than that of ED, Caput, Corpus and VD which is 0.1±0.1, 1.5±0.6, 1.9±0.7 and 0.3±0.2%, respectively (p<0.05). In experiment 3, percentage of intact plasma membrane spermatozoa of each regions were examined by hypoosmotic swelling test. Percentages of intact plasma spermatozoa were not significantly different among six regions of epididymis: ED, caput, corpus, Pcauda, Dcauda and VD which is 68.0±8.6, 74.0±5.3, 68.5±6.2, 70.8±5.5, 71.0±5.8 and 64.6±10.8%, respectively. In conclusion, in the present study, we found out distribution, motility and plasma membrane integrity of spermatozoa from six regions of epididymis in Hanwoo bull. These results will be contributed to basic research about spermatozoa transportation and characters in epididymis of bull.

Associations of Ultrasonographic Features with Scrotal Pain after Vasectomy

조승훈,민승기,Seung Tae Lee 대한비뇨의학회 2011 Investigative and Clinical Urology Vol.52 No.11

Purpose: Scrotal discomfort is a recognized complication of vasectomy, but the natural history and incidence of this problem are uncertain. The typical ultrasonographic changes after a vasectomy primarily include epididymal thickening and epididymal tubular ectasia with diminished blood flow in the epididymis. We prospectively studied the differences in the ultrasonographic features of the testis and epididymis between patients with and those without scrotal discomfort after vasectomy. Materials and Methods: We prospectively assessed pain scores in 178 men who underwent outpatient bilateral no-scalpel vasectomy at our institution between January 2009 and December 2010. At 2 months after vasectomy, we evaluated the postoperative scrotal pain questionnaire and scrotal ultrasonographic features for patients who returned for semen analysis. On the basis of the scrotal information, we investigated the potential relationships between scrotal pain or discomfort and scrotal ultrasonographic features of both testes and epididymides. Results: The average age of the 114 men was 36.3 years (range, 29 to 53 years). group 1 (n=23), which reported scrotal pain or discomfort, showed no significant mean differences in the maximal diameter of the head of the epididymis when compared with group 2 (n=91), who had no scrotal pain or discomfort. Also, the width of the body of the epididymis between the two groups showed no significant differences. Conclusions: There were no significant differences in ultrasonographic features according to the presence of chronic scrotal discomfort after vasectomy. Therefore, causes of scrotal pain other than obstruction may need to be considered after vasectomy. Purpose: Scrotal discomfort is a recognized complication of vasectomy, but the natural history and incidence of this problem are uncertain. The typical ultrasonographic changes after a vasectomy primarily include epididymal thickening and epididymal tubular ectasia with diminished blood flow in the epididymis. We prospectively studied the differences in the ultrasonographic features of the testis and epididymis between patients with and those without scrotal discomfort after vasectomy. Materials and Methods: We prospectively assessed pain scores in 178 men who underwent outpatient bilateral no-scalpel vasectomy at our institution between January 2009 and December 2010. At 2 months after vasectomy, we evaluated the postoperative scrotal pain questionnaire and scrotal ultrasonographic features for patients who returned for semen analysis. On the basis of the scrotal information, we investigated the potential relationships between scrotal pain or discomfort and scrotal ultrasonographic features of both testes and epididymides. Results: The average age of the 114 men was 36.3 years (range, 29 to 53 years). group 1 (n=23), which reported scrotal pain or discomfort, showed no significant mean differences in the maximal diameter of the head of the epididymis when compared with group 2 (n=91), who had no scrotal pain or discomfort. Also, the width of the body of the epididymis between the two groups showed no significant differences. Conclusions: There were no significant differences in ultrasonographic features according to the presence of chronic scrotal discomfort after vasectomy. Therefore, causes of scrotal pain other than obstruction may need to be considered after vasectomy.

Effects of vasectomy on epididymal functions and production of anti-sperm antibodies

Jae Eun Lee,Young Seok Oh,Woong Heo,Myung Chan Gye 한국발생생물학회 2013 한국발생생물학회 학술발표대회 Vol.2013 No.8

After vasectomy (VAX), the flux and composition of the epididymal fluid are modified, leading to sequelae to the epididymis. In an effort to understand molecular pathophysiology of the epididymis following VAX, we investigated the changes of gene expression and sperm functions in the epididymis of vasectomized mice. After VAX, the epithelial cell height was significantly decreased in cauda epididymis, resembling the those of vas deference. This suggests that VAX evokes alteration of segmental characteristics of epididymal epithelium. Of note, these was an increase in luminal diameter in corpus and cauda epididymis, indicating the alteration of fluid homeostasis in epididymal lumen and protein synthesis and secretion. Also, the formation of sperm granuloma and infiltration of inflammatory cells were noted in lumen of epididymis at 8 weeks postvasectomy, indicating the activation of immune response in epididymis. The serum TNF-α levels and epididymal TNF-α and IL-1β mRNA levels were significantly increased in VAX mice. Microarray analysis demonstrated that claudin (CLDN) 10 and cystic fibrosis transmembrane conductance regulator (CFTR) were downregulated after vasectomy. In contrast, angiotensin II receptor type 2 (Agtr2) was up-regulated after vasectomy. Taking into account the functional importance of angiotensin system in epididymal epithelia and muscle tissue, VAX may alter the secretory function of epithelia and sperm transport via alteration in angiotensin system. Importantly, the IgG type of anti-sperm antibody (ASA) were markedly increased in the blood of vasectomized mice. Together this indicates that VAX provokes local inflammation in epididymis as well as systemic inflammation, which in turn change the blood epididymal barrier, an important element for epididymal immunological privilege. In addition, the number of sperm in cauda epididymis was increased but the sperm motility was decreased after vasectomy. The spontaneous acrosome reaction was increased by vasectomy after capacitation. This suggests that VAX affects sperm functions as well as sperm maturation. In conclusion, bilateral vasectomy lead to local immune response of epididymis, causing immunologic infertility in men.

Expressional Modulation of Aquaporin 1 and 9 in the Rat Epididymis by an Anabolic-Androgenic Steroid, Nandrolone Decanoate

이기호 한국발생생물학회 2021 발생과 생식 Vol.25 No.4

The spermatozoa become mature in the epididymis which is divided into initial segment and caput, corpus, and cauda epididymis. The water movement across the epididymal epithelium is important for creating luminal microenvironment for sperm maturation. Aquaporins (Aqps) are water channel proteins, and expression of Aqps is regulated by androgens. The current research was focused to examine expressional regulation of Aqp1 and Aqp9 by an androgenic-anabolic steroid, nandrolone decanoate (ND). The ND at the low dose (2 mg/ kg body weight/week) or high dose (10 mg) was subcutaneously administrated into male rats for 2 or 12 weeks. Transcript levels of Aqp1 and Aqp9 were determined by quantitative real-time polymerase chain reaction (PCR) analyses. In the initial segment, level of Aqp1 was decreased with 12 week-treatment, while Aqp9 level was decreased by the high dose treatment for 12 weeks. In the caput epididymis, Aqp9 expression was decreased by the low dose treatment. The 2 week-treatment resulted in an increase of Aqp1 level but a decrease of Aqp9 expression in the corpus epididymis. In the corpus epididymis, the 12 week-treatment at the low dose caused the reduction of Aqp1 and Aqp9 levels, but the high dose treatment resulted in an increase of Aqp1 expression and a decrease of Aqp9 level. In the cauda epididymis, Aqp1 expression was decreased by 2 and 12 week-treatments, while increases of Aqp9 levels was detected with the high dose treatment for 2 weeks and with 12 week-treatment. These findings indicate differential regulation of Aqp1 and Aqp9 expression among epididymal segments by ND.

Effect of androgen for expression of aquaporin 5 in mouse epididymis

Jae Eun Lee,Chan Jin Park,Il Kyoo Koh,Myung Chan Gye 한국발생생물학회 2012 한국발생생물학회 학술발표대회 Vol.31 No.-

Aquaporin 5 (AQP5) implicated in the generation of saliva, tears, and pulmonary secretions functions as a water-specific channel. Epididymal epithelial cells actively reabsorb water, ions and proteins. Large quantity of testicular fluid movement across the epididymal tubules generates high osmotic milieu which is important for sperm maturation. In an effort to understand the fluid homeostasis and its regulation by sex steroids in male reproductive tract, the expression of AQP5 was examined in different regions of mouse epididymis during postnatal development. The effect of androgen on the expression of epididymal AQP5 was examined in ORX model. AQP5 mRNA levels were the highest in corpus region in which drastic increase was noted during sexual maturation. Epididymal AQP5 immunoreactivity was largely found in apical as well as basal region of luminal epithelia. Moderate immunoreactivity for AQP5 was found in in smooth muscle cells in both immature and mature mice. Epididymal lumen of ORX mice showed shrinkage together with decrease in AQP5 expression. Alteration of AQP5 expression in ORX epididymis was partially recovered by androgen injection. AQP5 mRNA was induced at 10uM 5α-DHT in organ cultured epididymis. Chromatin immunoprecipitatin (ChIP) showed that 5α-DHT induced recruitment of androgen receptor (AR) to the -4635 to -4453 bp region of the AQP5 gene promoter in adult epididymis. Taken together, axial regulatory mechanism may control transcription of AQP5 along the length of epididymal tubule. Water transport through AQP5 is important for late sperm maturation and storage in epididymis. Androgen may directly induce AQP5 gene transcription via activation of AR in epididymis.