• Journal of Genetic Medicine
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• v.1 no.1
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• pp.51-56
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• 1997

Spermatogenesis is known to be regulated by a number of genes and several factors such as hormones, growth factors, cytokines and others. This study was done to evaluate the relationship between HSPs and DAZ genes in human spermatogenesis; we observed the expression pattern of HSP gene in azoospermia men with DAZ gene that regulated the gene expression related with human spermatogenesis. RT-PCR method was used to detect DAZ, HSP70A, and HSP70B transcripts in all RNA samples. Total RNA was extracted from 21 testis tissues using TRIZOL reagent. cDNAs were synthesized with reverse transcriptase, AMV. All PCR reaction were performed on a PCR themocycler with DAZ, HSP70A, and HSP70B-specific primers. Semen analysis, karyotyping and testis histology were performed. DAZ gene, known as a candidate gene of azoospermia factor(AZF), was deleted in 2 of 21 patients. To evaluate the only effects of HSPs in this patients, 2 DAZ deleted cases were removed. We observed the mRNA of HSP70B in 5 whereas none could be seen with regard to HSP70A. Furthermore, the sperm of these 5 men were discovered to be immature. In conclusion, HSP70B as well ad DAZ gene seem to be involved causing spermatogenic failure. We suggest that HSP70B plays an important role in spermatogenesis and it is one of factors induced sperm maturation in human.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.12
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• pp.1557-1565
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• 2010

Methyl paraben (MP), which is used as a preservative in pharmaceutical and cosmetic (shampoo) products, foods and beverages, enters into the aquatic environment and can pose a potential fish health hazard. In this experiment, effects of MP were evaluated in adult male common carp (Cyprinus carpio) by exposing them to fractions (1/$143^{rd}$ to 1/29th) of the $LC_{50}$ dose with every change of water for 28 days. Vitellogenin induction, metabolic enzymes, somatic indices and bioaccumulation were studied at weekly intervals. The $96^{th}$ h $LC_{50}$ of MP in fingerlings was 120 mg/L. Compared to the control, except for increases (p<0.01) in alkaline phosphatase (EC 3.1.3.1), alanine aminotransferase (EC 2.6.1.2) and liver size, there were decreases (p<0.01) in activity of acid phosphatase (EC 3.1.3.2), aspartate aminotransferase (EC 2.6.1.1), and testiculosomatic index following exposure to any dose of MP. Vitellogenin induction was significantly higher (p<0.01) in exposed than unexposed (control) fish. The bioaccumulation of MP in testis, liver, brain, gills and muscle tissues of fish increased significantly (p<0.01) with increase of dose from 0.84 ppm to 1.68 ppm. Dose and duration of exposure (p<0.01) indicated that an exposure period of 1 to 2 weeks was sufficient to cause changes in the quantifiable parameters studied. Fish exposed to 4.2 ppm MP became lethargic after the $26^{th}$ d. Histologically, degeneration, vacuolization and focal necrotic changes in liver and fibrosis-like changes in testicular tissue were noted.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.3
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• pp.184-191
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• 2000

Gametogenesis and the reproductive cycle of the marsh clam, Corbicula leana(Prime} were investigated monthly Hyongsan estuary, Korea from January to December 1998 by histological observation. C. leana had separate sexes, and oviparous. The gonads were located between the sub-region of mid-intestinal gland in visceral cavity and reticular cennective tissue. The ovary and testis were composed of a number of ovarian and testicular tubules, respectively, Mature oocytes were characterized by germinal vesicles with nucleoli and their sizes ranged $70\;to\;80{\mu}m$ in diameter. A number of mesenchymal tissues and pigment granular cells were distributed in the growing oocytes and spermatocytes during early developmental stages. With the further development of gonad, these tissuse and cells gradually disappeared. The monthly changes in the fatness index were closely related to reproductive cycle. Minimun size for sexual maturity was reached over 10.0 mm in shell length. The reproductive cycle could be classified into 6 stages : multiplicative (March to April): early growing and growing (May to June), mature (July to August), spent (September), degenerative (October) and recovery (November to February).

• Anatomy and Cell Biology
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• v.56 no.4
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• pp.508-517
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• 2023

In cancer patients, chemo/radio therapy may cause infertility by damaging the spermatogenesis affecting the self-renewal and differentiation of spermatogonial stem cells (SSCs). In vitro differentiation of stem cells especially mesenchymal stem cells (MSCs) into germ cells has recently been proposed as a new strategy for infertility treatment. The aim of this study was to evaluate the proliferation and differentiation of SSCs using their co-culture with Sertoli cells and conditioned medium (CM) from adipose tissue-derived MSCs (AD-MSCs). Testicular tissues were separated from 2-7 days old neonate Wistar Rats and after mechanical and enzymatic digestion, the SSCs and Sertoli cells were isolated and cultured in Dulbecco's modified eagle medium with 10% fetal bovine serum, 1X antibiotic, basic fibroblast growth factor, and glial cell line-derived neurotrophic factor. The cells were treated with the CM from AD-MSCs for 12 days and then the expression level of differentiation-related genes were measured. Also, the expression level of two major spermatogenic markers of DAZL and DDX4 was calculated. Scp3, Dazl, and Prm1 were significantly increased after treatment compared to the control group, whereas no significant difference was observed in Stra8 expression. The immunocytochemistry images showed that DAZL and DDX4 were positive in experimental group comparing with control. Also, western blotting revealed that both DAZL and DDX4 had higher expression in the treated group than the control group, however, no significant difference was observed. In this study, we concluded that the CM obtained from AD-MSCs can be considered as a suitable biological material to induce the differentiation in SSCs.

• Development and Reproduction
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• v.14 no.4
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• pp.281-286
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• 2010

Recent evidence has revealed that the intratesticular injection of hypertonic saline(20%) resulted in a chemically castrated state such as nadir testosterone levels in rats. To confirm the efficacy of this simple saline-injection method further, we investigated the changes in the gross and microscopic anatomy of testis. Our study comprised three groups; intact(control) group, orchidectomy group and saline-injection (experimental) group. Single dose of hypertonic saline (sterilized, $750{\mu}{\ell}/testis$) were directly administered into both testis of adult rats (about 300 g BW). Bilateral orchidectomy was performed at the same day of saline injection. Following 30 days post-injection, reproductive tissues were surgically removed, weighed and fixed for histological examination. The body weights were not changed in both orchidectomy group and saline-injection group when compared to those in intact group. The wet weights of testis were significantly decreased in saline-injection group when compared to those in intact group. The wet weights of epididymis and seminal vesicle and prostate were significantly decreased in orchidectomy group and saline-injection group when compared to those in intact group. Macroscopically, the testes exerted slight atrophy and the tunica albuginea seemed to be intact in saline injection group. Histologically, however, larger parts of testicular tissue underwent necrosis and were barely recognizable after hematoxylin-eosin staining. In the same section, only the opposite part of the injection site was stained showing abnormal state of cell layers mostly fibrosis and infiltrated leukocytes. Sloughing of immature germ cells from the basement membrane along with shedding cells in the intraluminal space was notable in most seminiferous tubules from the saline injected testis. The present study confirmed that the direct injection of hypertonic saline into testis can induce a castration-like, testosterone-depriving effects on accessory sex organs. Our findings suggest that the efficacy of this less expensive and minimally invasive method seems to be almost even with that of conventional orchidectomy and chemical castration, though more in-depth evaluation should be supported.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.6
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• pp.563-574
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• 1986

The structure of gonads, gametogenesis and reproductive cycle of the jackknife clams, Solen strictus and Solen gordonis were investigated mainly by histological observation. The first species used were monthly sampled at the coastal area of Dadaepo, Pusan, Korea and Naechodo, Kunsan, Korea for one year from February 1982 to January 1983. The second species were monthly sampled at the sand beach of Dadaepo, Pusan, Korea, from February 1982 to January 1983. Sexualities of Solen strictus and Solen gordonis are dioecious, and these species are oviparous. The gonads are irregularly arranged from the subregion of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary was composed of a number of small ovarian sacs and the testis was composed of several testicular lobuli which from the tubular structure. Early multiplicating oogonium was about $10{\mu}m$ in diamater. Nucleus and nucleolus, at that time, were distinct in appearance. Each of the early growing oocytes made an egg-stalk, connected to the germinal epithelium of the ovarian sac. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed in the ovarian sacs in the early development stages. With the further development of gonad, these tissue and cells gradually disappeared. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells function as nutritive cells in the formation and development of the early stage germ cells. Mature oocytes were free in the lumen of ovarian sacs and gradually become round or oval. Ripe oocyte was about 80 to $90{\mu}m$ in diameter. With the further development of testis, each of the testicular lobuli formed stratified layers composed of spermatogonia, spermatocytes, spermatids and spermatozoa in groups on the germinal epithelium. After spawning, the gonad gradually degenerated, and disorganized completely. Then new differentiated tissues were rearranged next year. The annual reproductive cycle of those species could be classified into five stages; multiplicative, growing, mature, spent, degenerative and resting stage. It seems that the spawning season is closely related to the water temperature, and the spawning of Solen strictus occurs from June to July at above $20^{\circ}C$ in water temperature. The peak spawning season appeared in June at Dadaepo and in July at Kunsan, The spawning of Solen gordonis occurs from May to June with the peak spawning season in June. Percentages of the first maturity in female of Solen strictus ranging from 5.1-6.0 cm and 7.1-8.0 cm in shell length were $50\%$ and $100\%$, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.6
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• pp.835-841
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• 2018

Objective: The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor (KISS1) gene vaccine in immunocastration. Methods: Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected. Results: The specific anti-KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p<0.05). No significant difference was observed at weeks 22 and 30 after the primary immunization. Similar results were also found for scrotal circumference, testicular weight, length, breadth, and spermatogenesis in seminiferous tubules in week 30 after primary immunization. KS (KISS1-hepatitis B surface antigen S) fusion fragment of KISS1 gene vaccine was not detected in host cell genomic DNA of 9 tissues of the vaccinated ram lambs by polymerase chain reaction. Conclusion: The effects of KISS1 gene vaccine in immunocastration were reversible and no integration events were recorded.

• Development and Reproduction
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• v.12 no.3
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• pp.215-220
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• 2008

Sex differentiation process of the spotted Black Bullhead, Pseudobagrus koreanus, was investigated using fish samples of different age after hatching. The primordial germ cells appeared separately under air bladder in 1-day larva (total length: $6.63{\sim}6.95\;mm$). The primordial gonad with a genital ridge developed in 5-day prelarva ($7.50{\sim}9.36\;mm$). The ovarian differentiation started in about 25-day juvenile ($11.58{\sim}13.21\;mm$). The somatic tissues enlongated in the tip of one end of undifferentiated gonad and fused each other. Thus a small ovarian cavity appeared. The testicular differentiation was initiated in 30-day juvenile ($12.19{\sim}13.72\;mm$). The rudiment of sperm duct was appeared in the lower part of the undifferentiated gonad. In 50-day juvenile ($16.28{\sim}17.06\;mm$), the ovary started to fill with peri-nucleolus oocytes, and the spermatogonia started to develop. In 250-day juvenile ($35.49{\sim}51.12\;mm$), the ovary became bigger and filled with oocytes, and the number of spermatogonia started to increase. Considering these results, the spotted Black Bullhead could be a differentiated type in sex differentiation and gonochorism in sexuality.

• Korean Journal of Environmental Biology
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• v.20 no.3
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• pp.224-231
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• 2002

Photoperiod (length of light per day) is a major factor in regulating reproductive function in golden hamsters. The information of photoperiod is transmitted to the reproductive endocrine system by melatonin. Thus the effects of melatonin aye investigated in male golden hamsters exposed to photoperiods. Paired testicular weights were markedly reduced in the animals housed in short photoperiod $(SP,\le{12\;hours\;day^{-1})$ and injected with melatonin in the evening, but not in long photoperiod $(LP,\le{12.5}\;hours\;day^{-1})$ and injected with melatonin in the morning. The histological examination of regressed testes showed reduction of tubular lumen diameter including the numbers of cells and Leydig cell number. The mean values of both follicle stimulating hormone (FSH) and luteinizing hormone (LH) were also lowered in the sexually inactive animals than in the sexually active animals. Melatonin receptor was identified by reverse-transcription polymerase chain reaction (RT-PCR) and its expression was examined in various tissues to scrutinize the action site of melatonin. It turned out 309 nucleotides and was definitely expressed in hypothalamus and pituitary including spleen, retina, and epididymis. And gonadotropin releasing hormone (GnRH) gene, which is a key element in regulating reproduction, was identified by RT-PCR but the expression of GnRH was not modified by the treatment of melatonin. Taken together, photoperiod via melatonin indirectly affects reproductive endocrine system, possibly through the release of GnRH, not the synthesis of GnRH.

• Development and Reproduction
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• v.4 no.2
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• pp.195-201
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• 2000

Sex differentiation process of the spotted sea bass, Lateolabrax maculatus, was investigated by histological method. The fish samples were collected from just after hatching to 365 days later. The primordial germ cells and genital ridge were appeared separately hanging under air bladder in 30-day larva (total length: 11.7~13.2 mm), and were unified into the undifferentiated gonads in 40-day larva (12.5~14.0 mm). The ovarian differentiation was started in 60-day juvenile (23.6~27.0 mm). The somatic tissues were elongated in tip of both ends of undifferentiated gonad and were fused each other. The complete ovarian cavity was appeared in 80-days juvenile(33.1~42.5 mm). The testicular differentiation was initiated in 70-day juvenile (24.8~31.6 mm). The rudiment of sperm duct was appeared in the center of the undifferentiated gonad. The meiosis of germ cells in the ovary was started in 168-day juvenile (88.0~115.4 mm). In 287-day juvenile (175.1~233.6 mm), the ovary was filled with both of chromatin stage and perinucleolus stage oocytes. The meiosis of male germ cells was started in 245-day juvenile (124.4~168.3 mm). However, the seminiferous tubules of testis were filled with numerous sperm in 365-day juvenile (162.5~253.8 mm). The sex ratio of male and female was 1:1.38. Considering these results, the spotted sea bass was showed differentiated type in sex differentiation and gonochorism in sexuality.