• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.53-61
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• 1996

This study was carried out to determine effectively the sex of rabbit embryos using H-Y antiserum. H-Y antiserum was obtained from inbred SD strain female rat which was immunized by injection of testis cell of inbred SD strain male rate into its spleen. The titer of antiserum was identified by sperm cytotoxicity test and culture of rabbit embryos with antiserum. The developed or undeveloped embryos were separated by exposure the embryos to the antiserum with H-Y antibody. Developed embryo were transferred to the recipients and sex of offspring were examined. 1. In the sperm cytotoxicity test, the rate of dead sperm showed no difference between two antisera from spleen and testis cell as antigens. It is confirmed that H-Y antibody in antiserum was absorbed by H-Y antigen in male rat spleen cells. 2. When rabbit morulae were exposed to antiserum and complement, the rate of embryos developed or arrested was 51 and 49% respectively and the rate was closely same as natural sex ratio of 50:50%. 3. When rabbit morulae were cultured for 12h in the medium containing antiserum produced by antigen of testis cell, the rate of embryos developed or arrested was 48 and 52% respectively and the rate was closely same as natural sex ratio of 50:50%. 4. Eighty rabbit embryos which were not affected by H-Y antiserum were transferred to four recipients. Two recipients were pregnant and born 13 pups among which 2 (14%) were male and 11 (86%) were female. In conclusion, existence of H-Y antibody in the serum from female rat immunized by injecting testis cell from newborn male rat to the spleen of the female rat was confirmed. When rabbitmorulae were exposed to H-Y antiserum and complement, about a half of embryos were developed to blastocysts. When the rabbit embryos not affected by H-Y antiserum were transferred, the rate of female offspring was 86%. Therefore, it was identified that most of embryos which were not affected by H-Y antiserum were female.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.5
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• pp.658-671
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• 2018

Objective: Nandrolone decanoate (ND) is an anabolic-androgenic steroid frequently used for clinical treatment. However, the inappropriate use of ND results in the reduction of serum testosterone level and sperm production. The suppressive effect of ND on testosterone production has not been investigated in detail. The present study was designed to examine the effect of ND on the expression of steroidogenic enzymes in the rat testis. Methods: Male Sprague Dawley rats at 50 days of age were subcutaneously administrated with either 2 or 10 mg of ND/kg body weight/week for 2 or 12 weeks. The changes of transcript and protein levels of steroidogenic enzymes in the testis were determined by real-time polymerase chain reaction and western blotting analyses, respectively. Moreover, immunohistochemical analysis was employed to determine the changes of immunostaining intensity of these enzymes. The steroidogenic enzymes investigated were steroidogenic acute regulatory protein, cytochrome P450 side chain cleavage enzyme, $17{\alpha}-hydroxylase$, $3{\beta}-hydroxysteroid$ dehydrogenase, and cytochrome P450 aromatase. Results: The treatment of ND resulted in depletion of Leydig cells and sloughing of germ cells in the testis. The ND treatment caused significant expressional decreases of steroidogenic enzymes at transcript and protein levels, and the destructive effects of ND on the testis were more apparent with a higher dose and a longer period of the treatment. Evident reduction of immunostaining intensity present in Leydig cells was clearly detected by the ND treatment. Conclusion: The exposure to ND in young male results not only in histological changes of the testis but also in aberrant gene expression of testicular steroidogenic enzymes, consequently leading into the reduction of testosterone production in the testis and thus likely disruption of spermatogenesis.

• Applied Microscopy
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• v.29 no.3
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• pp.353-362
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• 1999

Di-(2-ethylhexyl) phthalate (DEHP) is a plasticize. known as one of endocrine disruptors. The present study was carried out to investigate the ultrastructural changes of prepubertal rat testis after oral administration of DEHP in dosages of 1 g/kg, 3 g/kg or 5g/kg in 0.5 ml of corn oil daily for a week. This study revealed the DEHP inhibited the development of seminiferous tubules and induced structural changes on various cell types of the rat testis. Leydig cells, Sertoli cells and the developing germ cells seemed to be impaired their differentiations in terms of the structural changes of cell organelles. The increase of heterochromatin in amount were common features in all 3 cell types. In addition, the Leydig cells were characterized by the increases in number and size of lysosomes and the scantiness of smooth endoplasmic reticulum. The Sertoli cells became irregular in nuclear envelope and the cytoplasm decreased, but the number of lysosomes and vacuoles seemed to be increased. There were some indications of necrosis of the germ cells, such as vacuolized nucleus and segregated nucleolus. These detrimental effects of DEHP on the rat testis were dose dependent and suppressed spermatogenesis decreasing developing germ cells in number and appearances. The effect of DEHP on ultrastructure of rat testis, as its known physiological functions, seems come from the decreased level of testosterone by Leydig cells, followed by the abnomalities of Sertoli cells and the germ cells.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.531-539
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• 2003

Changes in the rat testis interstitium from birth to adulthood were studied using Sprague Dawley rats of 1, 7, 14, 21, 28, 40, 60, and 90 days of age to investigate Leydig cell differentiation. In addition, serum testosterone concentrations and luteinizing hormone stimulated (LH; 100 ng/ml) testosterone secretory capacity per testis in vitro were determined via radioimmunoassay. Fetal Leydig cells were present in rat testes from birth to 21 days, and they were only steroidogenic cells in the testis at days 1 and 7. The average volume of a fetal Leydig cell and the absolute volume of fetal Leydig cell per testis were similar at all ages of experimental groups except at day 21 when lower values were observed for both parameters. The number of fetal Leydig cells per testis remained constant from birth through 21 days. Adult Leydig cells were recognized at day 14 and their absolute volume and number per testis increased linearly from 14 to 90 days. The average volume of an adult Leydig cell increased significantly with age and reached maximum size by 60 days of age where the volume was nearly three times bigger than that of at day 14. Total testosterone production per testis in vitro and serum testosterone concentrations were not significantly different at day 1 compared with 7, 14, and 21 days of age. Significant increases were observed at days 40 and 60. Values at days 60 and 90 were not significantly different.

• Environmental Analysis Health and Toxicology
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• v.21 no.3 s.54
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• pp.291-299
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• 2006

The experiments investigated whether early exposure to testosterone propionate (TP) during prepuberty alters testis development in Sprague-Dawley male rats. We performed Hershberger assay using the stimulated weanling male rats by OECD protocols, cDNA microarray, and Western blot. TP was subcutaneously injected to uncastrated Sprague-Dawley male rat of 22 days old for 10 consecutive days at doses of 0.4, 0.8, 1.0, 1.2, 1.6 mg/kg per day. At necropsy, the following tissues were removed and weighed: combined testes, epididymides (Epi), Cowper's glands (COW), levator am, and bulbocavernosus muscles (LABC), seminal vesicles, together with coagulating gland (SV) and ventral prostate (VP). We found that TP increased the weights of Epi, VP, SV, COW, and LABC, while testis was decreased in a dose-dependent manner. In cDNA microarray analysis of testis, there were significant reductions in the expression of cytochrome P450 11A (CYP11A), the rate-limiting enzyme of steroidogenesis. Taken together these results, TP exposure before puberty in male rats may produce the delay in testis development by inhibiting the CYP11A gene expression.

• The Journal of Korean Obstetrics and Gynecology
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• v.28 no.3
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• pp.54-73
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• 2015

Objectives The object of this study was to evaluate the effect of Yikgeebohyul-tang aqueous extracts (YKBHT) on the propylthiouracil (PTU)-induced rat hypothyroidism. Methods The rats were divided into 6 groups : intact vehicle control, PTU control, LT4, YKBHT 500, 250 and 125 mg/kg treated groups. Hypothyroidism was induced by daily subcutaneous treatment of PTU 10 mg/kg for 28 days. YKBHT aqueous extracts were administered once a day as an oral dose of 500, 250 and 125 mg/kg for 42 days. The changes were observed : weight of body, thyroid gland, liver, testis, epididymis and prostate, serum thyroid hormone levels, serum male sex hormone levels, serum lipid profiles, liver and testis antioxidant system. These results were compared with LT4 0.5 mg/kg intraperitoneally treated rats. Results These PTU induced hypothyroidism and related hepatic and male reproductive organ damages were favorably and dose-dependently inhibited by treatment of YKBHT 500, 250 and 125 mg/kg, and YKBHT also effectively regulated the PTU-induced abnormal antioxidant defense factor changes in the both liver and testis. Although, LT4 also inhibited PTU-induced hypothyroidism and relative liver damages. But it deteriorated the hypothyroidism related testis, epididymis and prostate damages through testicular oxidative damages. Conclusions : The result of this study suggests that YKBHT has favorable effects on the hypothyroidism and related liver and reproductive organ damages with augmentation of antioxidant defense factor in the testis and liver. YKBHT 500, 250 and 125 mg/kg dose-dependently inhibited PTU-induced hypothyroidism and related liver and male reproductive organ damages in rats.

• Radiation Oncology Journal
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• v.8 no.1
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• pp.17-27
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• 1990

The effects of both hyperthermia alone and X-ray irradiation combined with hyperthermia on rat testis have been investigated. The histological changes were observed on 15 and 30 days after treatment. There was no histological change of rat testis by hyperthermia alone. The earliest change by X-ray irradiation was the degeneration of the spermatogonia of the seminiferous tubule, which was appeared in 2 Gy group. Necrosis of the spermatogonia was severe in 6 Gy group and complete atrophy was developed in 8 Gy group. With increased dose of radiation, the degree of changes of tubules was increased. In combined group of X-ray irradiation and hyperthermia, the histological change of the seminiferous tubule was more severe than X-ray alone group. Necrosis and atrophy of the spermatogonia were appeared in 2 Gy and complete atrophy of spermatogonia was seen in 6 Gy group. Thermal enhancement ratio (calculated at the complete atrophy of the spermatogonia) was 1.3 in this experiment. There was no difference in observation time inverval between 15 and 30 days after each treatment in all groups.

• Journal of Animal Science and Technology
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• v.53 no.3
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• pp.195-202
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• 2011

Maintenance of adequate telomere length in developing cells is the most important concern to preserve the integrity of the genome. The length of telomere is strictly regulated by numerous telomere-binding proteins and/or interacting factors. Even though the expression of telomerase in the male reproductive tract has been characterized, developmental expressional profiling of telomerase and other telomere-associated proteins has not been determined in detail. The present study was attempted to examine expression patterns of catalytic subunit (Tert) and RNA component (Terc) of telomerase and two telomerase associated factors, telomerase associated protein 1 (Tep1) and TERF1 (TRF1) interacting nuclear factor 2 (Tinf2) in the testis and seminal vesicle of male rat during postnatal development. The real-time PCR analysis was utilized to quantify mRNA expression of molecules. The abundance of Tep1 mRNA in the testis and seminal vesicle was the highest at 5 months of age. Expressional fluctuation of Tinf2 during postnatal development was found in the testis, while expression of Tinf2 in the seminal vesicle was gradually increased until 5 months of age and then significantly decreased later. mRNA level of Tert gene in the testis was significantly increased at the adult and the elder, while the highest expression of Tert gene in the seminal vesicle was found at 5 months of age. Expression of Terc transcript in the testis and seminal vesicle was the highest at 5 months of age, followed by significant reduction at 1 and 2 years of ages. Such differential gene expression of telomere-associated factors and telomerase components in different male reproductive tissues during postnatal development indicates that maintenance of telomere length would be regulated in tissue- and/or age-specific manners.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2006.10a
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• pp.89-90
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• 2006

• Applied Microscopy
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• v.16 no.2
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• pp.75-91
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• 1986

Differentiation of the rat testis was studied by light and electron microscope from the fetal stage up to the newborn or adult stage. The purpose of the present study is to investigate the ultrastructural changes of seminiferous tubules and interstitial tissue during the developmental process. The results were as follows: the seminiferous tubule diameter began to increase from birth and was fully developed at 30 to 40 days of age through intratubular cell proliferations. Basement membrane and myoid cells lining the seminiferous tubules were differentiated at 17 days gestation. At the fetal stage, seminiferous tubules were primarily composed of Sertoli cells and the differentiation of Sertoli and germ cells progressed from the newborn stage. Spermatids and immature spermatozoa are appeared at 40 days of age, so from this time, spermatogenesis occurred actively until the adult stage. Sertoli cells aided germ cell differentiation and phagocytosed the parts of the spermatid cytoplasm. Leydig ce]] development follows a biphasic pattern: a fetal phase and then an adult phase from 20 days of age. In conclusion, the rat testis is already developed to some extent by the fetal stage and is functional after 50 days of age. Therefore, these findings indicate that differentiation of Sertoli and Leydig cells precedes the onset of spermatogenesis.