• Asian-Australasian Journal of Animal Sciences
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• v.31 no.2
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• pp.189-197
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• 2018

Objective: Hemicastration is a unilateral orchiectomy to remove an injured testis, which can induce hormonal changes and compensatory hypertrophy of the remaining testis, and may influence spermatogenesis. However, the underlying molecular mechanisms are poorly understood. Here, we investigated the impact of hemicastration on remaining testicular function. Methods: Prepubertal mice (age 24 days) were hemicastrated, and their growth was monitored until they reached physical maturity (age 72 days). Subsequently, we determined testis DNA methylation patterns using reduced representation bisulfite sequencing of normal and hemicastrated mice. Moreover, we profiled the testicular gene expression patterns by RNA sequencing (RNA-seq) to examine whether methylation changes affected gene expression in hemicastrated mice. Results: Hemicastration did not significantly affect growth or testosterone (p>0.05) compared with control. The genome-wide DNA methylation pattern of remaining testis suggested that substantial genes harbored differentially methylated regions (1,139) in gene bodies, which were enriched in process of protein binding and cell adhesion. Moreover, RNA-seq results indicated that 46 differentially expressed genes (DEGs) involved in meiotic cell cycle, synaptonemal complex assembly and spermatogenesis were upregulated in the hemicastration group, while 197 DEGs were downregulated, which were related to arachidonic acid metabolism. Integrative analysis revealed that proteasome 26S subunit ATPase 3 interacting protein gene, which encodes a protein crucial for homologous recombination in spermatocytes, exhibited promoter hypomethylation and higher expression level in hemicastrated mice. Conclusion: Global profiling of DNA methylation and gene expression demonstrated that hemicastration-induced compensatory response maintained normal growth and testicular morphological structure in mice.

• The Korean Journal of Malacology
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• v.29 no.2
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• pp.105-111
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• 2013

The gonadal development of triploid and diploid Pacific abalones, Haliotis discus hannai was histologically investigated in spawning season. Diploid abalones had matured oocytes and spermatozoa, but most triploid had spermatocytes or developing oocytes that was slightly retarded in gonadal development compared to diploid abalones. In spawning experiment of triploid and diploid abalones, spawning rates of diploid male and female were 100%, but those of triploid female was 50% and male was 25% respectively. Investigation of spawned abalone eggs and spermatozoa revealed that length of diploid sperms head were 17.47 ${\mu}m$, breadth of head were 10.31 ${\mu}m$, length of spermatozoa were 130.72 ${\mu}m$, but those of triploid spermatozoa were 11.83 ${\mu}m$, 7.89 ${\mu}m$ and 103.36 ${\mu}m$ respectively. Triploid spermatozoa were significantly small to diploid spermatozoa (p < 0.05). The eggs of diploid and triploid were not different in size. The cross experiment between oocytes produced by triploids and spermatozoa by diploids ($3n{\times}2n$ cross) revealed that no fertilization were occurred, and $2n{\times}3n$ cross also revealed same result.

• Journal of Acupuncture Research
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• v.23 no.1
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• pp.105-120
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• 2006

To investigate anti-cancer effects of wild ginseng herbal acupuncture and mitigation of anti-cancer drug when taken concurrently, cancer cells from B16/F10 melanoma were injected intraperitoneally in C57BL/6. After inducing cancer, anti-cancer effects and mitigation of reproductive toxicity of Doxorubicin were evaluated. 1. For changes in weight, Doxorubicin treated group showed significant decrease, and administration of wild ginseng herbal acupuncture didn't cause any weight change. 2. Volume of tumor was significantly reduced in Doxorubicin teated group. Wild ginseng herbal acupuncture groups showed slight decrease but insignificant compared to the control group. 3. For hematological evaluation, Doxorubicin only group's reticulocytes were significantly decreased compared to the control group, and Platelet Count was significantly increased. Wild ginseng herbal acupuncture group showed significant increase of Neutrophils and significant decrease of Lymphocytes compared to the control group. 4. For histological evaluation of the tumor, necrosis occurred in a wide range in the Doxorubicin treated group. Wild ginseng herbal acupuncture didn't cause much histological changes. 5. For histological evaluation of the testis, seminiferous tubules of the control group suffered severe damage on epithelial cells. When wild ginseng herbal acupuncture was administered concurrently, damage on the seminiferous tubules was significantly inhibited compared to the Doxorubicin only group. 6. Diameter of seminiferous tubules and spermatogonia count were insignificant between the experiment groups. 7. For BrdU positive reaction of testicle tissue, Doxorubicin only group failed to show any reaction of spermatogonia, but spermatocytes and spermatids showed slight positive reaction. When wild ginseng herbal acupuncture was treated concurrently, much greater positive reaction was made but similar to that of the control and normal groups. 8. For observation of changes in BrdU spermatogonia count of the testicle tissue, Doxorubicin only group didn't show any positive reaction, and relative increase was shown in the group with concurrent administration of wild ginseng herbal acupuncture. 9. For observation of TUNEL positive reaction cells of the testicle tissue, no significant changes were witnessed in all the experiment groups.

• Development and Reproduction
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• v.2 no.1
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• pp.89-99
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• 1998

This study was carried out to investigate the morphological changes of gonadotropes in pituitary gland and spermatogenic cells in testis, obtained from 150 of 3-year-old immature and mature male rainbow trout (Oncorhynchus mykiss) during the reproductive cycles from March to February in the following year. In the maturation cycle of the pituitary gonadotropes of cultured rainbow trout, three periods can be distinguished i.e. a period of resting(March-August), a period of full spermatogenesis (September-November), and a period of breeding (December-February). The ultrastructures of the gonadotropes largely parallel the cyclical changes in the tests. The seminiferous tubules contain all spermatogenetic stages and sperm cells in a period of early maturation. At first, the size of the nucleus and cytoplasm decrease gradually at every stages from spermatogonia to spermatids. In the secondary spermatocytes, the small mitochondria are located over the outer cytoplam. In spermatids, the cytoplasmic masses move toward the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm. In spermatids, the cytoplasmic masses move towark the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm and begin to elongate. In spermatozoa, the surface of the nucleus devreases in volume. Examination by TEM shows that the nuclear envelope and plasma membrane are slightlywrinkled and closely adhered to the nucleus of spermatozoa. Two oval mitochondria are quite separated and the flagellum is inserted into the base of the spermatozoa head.The axoneme in this fish has the typical pattern such as nine peripheral doublets and a central doublet(9+2). there are remarkable individual differences in the size and morphology of spermatozoa head as observed by transmission and scanning electron microscopy.

• Development and Reproduction
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• v.20 no.4
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• pp.289-296
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• 2016

This report describes the sex differentiation of the Korean rose bitterling, Rhodeus uyekii, from hatching to 170 days post-hatch (DPH) in relation to total length (TL), body weight (BW), and integral water temperature (IWT). The growth curve of TL from just hatching to 83 DPH was $5.144e^{0.045t}$ ($R^2=0.961$; t, time), and that of BW was $2.398e^{0.086t}$ ($R^2=0.725$). Primordial germ cells (PGCs) were observed at 17 DPH (7.9 mm TL, 3.74 mg BW, $374^{\circ}C$ IWT), and thereafter began to protrude into the peritoneal cavity. At 21 DPH ($9.2{\pm}0.14mm$ TL, $4.8{\pm}0.07mg$ BW, $462^{\circ}C$ IWT), some PGCs contained condensed chromatin and oocyte were observed in meiotic prophase. In contrast to the ovaries, which grew gradually after sexual differentiation, testes began multiplying at 25 DPH (10.1 mm TL, 5.42 mg BW, $550^{\circ}C$ IWT), when testicular differentiation was first identified, and multiplied continuously thereafter. At 33 DPH (11.2 mm TL, 10.5 mg BW, $726^{\circ}C$ IWT), the developing testes contained spermatogonia that exhibited mitotic activity. No spermatocyte or sperm cell was observed until 83 DPH (18.9 TL, 48.2 mg BW, $1,826^{\circ}C$ IWT). At 170 DPH (32.5 mm TL, 270.1 mg BW, $3,740^{\circ}C$ IWT), which was the end point of this study, the mature ovaries showed germinal vesicle breakdown, while the mature testes contained observable spermatocytes and sperm cells. These results allow us to identify the sex differentiation type of the Korean rose bitterling as differentiated gonochoristic.

• Applied Microscopy
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• v.33 no.1
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• pp.25-39
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• 2003

Cell differentiation and ultrastructural characteristics in the seminiferous epithelium of Myotis macrodactylus was investigated with the light and electron microscopes. Spermatogenesis has begun at April and finished at September. The nuclei of A spermatogonia (dark and pale type of spermatogonia) were oval, applied to the basal lamina, and surrounded by Sertoli cells. By comparison with other types of spermatogonia, the cell and nucleus of B type of spermatogonium is globular and larger than A types of spermatogonia. The nucleolus appears as a coarse and touches the nuclear membrane. The cell and nucleus of spermatocytes was globular and larger, but primary spematocyte is larger than secondary spermatocyte. Spermiogenesis was divided according to the level of fine structural difference, into Golgi, cap, acrosomal, maturation and spermiation phases; Golgi, cap, acrosomal and spermiation phases were further subdivided into steps of early and late phase respectively, and maturation phase has only one step. Hence, the spermiogenesis has been divided into a total of nine phases. In the change of karyoplasm, the chromatin granules are condensed at late Golgi phase and completed at spermiation phase. The sperm tail began to develop in early Golgi phase and completed in spermiation phase. The process of degeneration of spermatogenic cells in the seminiferous tubules was continually observed from October, before the beginning of hibernation, to hibernation phase (November, December, January, February, March). Immatured spermatogenic cells in the seminiferous tubules have been engulfed by phagocytosis of Sertoli cells during period of degeneration. It is deduced that the adaptative strategy serves as the mechanism to regulate the effective use of energy to prepare for long hibernation and regulation of breeding cycle.

• Applied Microscopy
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• v.40 no.3
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• pp.169-176
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• 2010

Black molly (Poecilia sphenops) and sailfin molly (Poecilia latipinna) are a teleost belonging to Poeciliidae. The spermatogenesis between two species were investigated by light and electron microscope. The whitish testes of both black molly and sailfin molly were located between intestine and air bladder. The size of testis was major axis 7 mm, minor axis 2 mm. The testis contained numerous testicular cysts. In both black molly and sailfin, primary spermatocytes were comparatively large ellipsoidal, and mitochondria showed a marked development. The secondary spermatocyte was smaller than that of primary spermatocyte, highly condensed according to their development. The nucleus with electron-dense was round shape and flagella started to be formed. In spermiogenesis, chromatin was more condensed. The mitochondria were rearranged along the tail. The number of mitochondria was 2 to 4 in cross section and 8 to 10 in longitudinal section. The head of mature sperm was long cone shape and had not acrosome. The microtubules of flagella were arranged 9+2 structure. Also, the tail of sperm have lateral fins. In conclusion, spermatogenesis and sperm morphologies of these two species were same. These morphological similarity seems to be an indication of the Poeciliidae.

• Korean Journal of Animal Reproduction
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• v.4 no.1
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• pp.47-74
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• 1980

The present experiments were carried out to elucidate the effects of hypophysectomy and subsequent administration of sex hormone on thyroid, adrenal gland, gonads and blood plasma components in the rat. The jresults obtained were summarized as follows: The weight of the thyroid gland of both male and female hypophysectiomzed rats decreased markedly from 7 days up to 56 days after the hypophysectomy as compared to the control group. The administration of sex hormone (6 mg of testosterone propionate to male and 6 mg of hexestrol to female) to the hypophysectomized rat gave on effect on the change in the weight of the thyroid gland. The hopophysectomy decreased the uptake of radioactive iodine in the thyroid gland in both male and female rats with time. Subsequent administration of the sex hormone caused no effect. With regard to the histological changes of the thyroid gland, the hypophysectomy caused significant changes in the gland showing a remarkable degeneration. The function of the gland seemed to disa, pp.ar almost completely on 56th day after the hypophysectomy. Upon the administration of sex hormone after the hypophysectomy, however. the epithelia of the follicle which has changed to flat from has partly returned to its functional cubicfrom and nuclei recovered as nearly as normal. These recovery were more remarkable in the female than in the male. The hypophysectomy kept causing a significant decrease in the weight of the adrenal gland in male and female rats during the period of observation (up to 56 days) as in the case of thyroid gland. The administration of sex hormone has on effect in this respect either. The hypophysectomy also caused a marked morphological change in the gland: zona fasciculata and zona reticularis were dicreased in size quichly after the hypophysectomy. The administraton of the sex hormone to the hypophysectomized rat resulted in clear distinction among the three layers of the adrenal cortex which otherwise very diffused. In the male, this phenomenum was more remarkable than in the female and the pattern of the cell arrangements and the thickening of each layer became similar to those of normal rats. The gonads of both sexes have also kept decreasing in the weight and degenerated in morpohology after the hypophysectomy. However, the degenerate follicle became enlarged after the administration of hexestrol in the female. Furthermore, the vacuoles found in interstitial cells of hypophysectomized rat disa, pp.ared after the administration of testosterone in the male and the formation of spermatocytes seemed to be recovered. Hypophysectomy also caused a gradual increase in the contents of total protein, non-protein nitrogen, total lipid, cholesterol and calcium in the blood plasma with time. The concentrations of sodium, potassium and chloride in the blood did not change after the hypophysectomy. Sex hormone caused practically no change in above tendency.

• Animal cells and systems
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• v.16 no.2
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• pp.127-134
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• 2012

We investigated the effects of fadrozol, an aromatase inhibitor (AI), and $17{\alpha}$-methyltestosterone (MT) on the induction of sex change in juvenile longtooth grouper $Epinephelus$ $bruneus$, via histological observation of gonads. Changes in the mRNA expression of GtH subunits (FSH-${\beta}$ and LH-${\beta}$) in the pituitary, and estradiol-$17{\beta}$ (E2) and 11-ketotestosterone (11-KT) levels in the blood were also surveyed after AI and MT treatment. Juvenile longtooth groupers ($113{\pm}17g\;body\;weight$; $16.2{\pm}1.2cm\;body\;length$) received intramuscular injections of AI at 3 (3-AI) and 5 (5-AI) mg/kg BWdoses and MT at a 5 mg/kg BW (5-MT) dose. At week 7 post-injection, 3-AI and 5-MT oocytes were degenerated, and gonads of the 5-AI group initiated spermatogenesis. At week 21 post-injection, 3-AI- and 5-MT-treated gonads contained spermatogonia and spermatocytes, while 5-AI treatment induced advanced stages of spermatogenesis. The serum E2 level showed no significant differences throughout the experimental period, whereas that of 11-KT was significantly elevated in the 5-AI group at weeks 7 and 21 post-injection. A significant increase in the expression of FSH-${\beta}$ mRNA was evident in the 5-AI group at week 21 post-injection. In contrast, LH-${\beta}$ mRNA expression did not significantly differ among groups during the experimental period. These results imply that sex change has two stages in the longtooth grouper. In the first stage, oocytes are degenerated by the stimulation by 11-KT, and in the second stage spermatogenesis occurs, owing to the co-effects of 11-KT and FSH-${\beta}$.

• Journal of Veterinary Clinics
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• v.27 no.5
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• pp.514-521
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• 2010

Epimedium koreanum nakai (EKN) has been used for treatment of reproductive disorders. In this study, we evaluated the effects of EKN water extract on the 2-bromopropane (2-BP) induced reproductive dysfunction. The daily sperm production (DSP), sperm counts and histological changes of reproductive organs were investigated after 4 weeks of EKN administration in 2-BP induced reproductive damaged rats. Although the weights of epididymis and seminal vesicle were increased dose-dependently, they were lower than control group. DSP of EKN-dosing groups were increased compared to control group. In EKN-dosing groups, the number of degenerative seminiferous tubules and the number of epididymal tubules showing epithelial cell vacuolation and decreased spermatozoa in the lumen were significantly (p < 0.01) decreased compared to control group dose-dependently. Significant (P < 0.05 or P < 0.01) increases of the number of sertoli cells, spermatogonia, patchytene spermatocytes, round spermatids and elongated spermatids were observed in EKN-dosing groups compared control group. Consequently, based on the results, EKN water extract could treat 2-BP induced reproductive damages dramatically.