• The Korean Journal of Physiology and Pharmacology
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• v.25 no.4
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• pp.341-354
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• 2021

Cardamonin (CARD) is a chalconoid with anti-inflammatory and antioxidant properties, and it is present in several plants. We sought to explore whether CARD exerts any positive effects against hyperglycemia-induced testicular dysfunction caused by type 2 diabetes and aimed to identify its possible intracellular pathways. Adult male rats were subdivided into six groups: control, CARD, diabetic (DM), DM + glibenclamide (GLIB), DM + CARD and DM + GLIB + CARD. Type 2 DM induced a significant increase in blood glucose and insulin resistance, along with diminished serum insulin, testosterone and gonadotropins levels, which were associated with the impairment of key testicular androgenic enzymes and cellular redox balance. Administration of CARD at a dose of 80 mg/kg for 4 weeks effectively normalized all of these alterations, and the improvement was confirmed by epididymal sperm analysis. After treatment with CARD, the pathological changes in spermatogenic tubules were markedly improved. Significantly, CARD upregulated testicular glucose transporter-8 (GLUT-8) expression and had inhibitory effects on elevated autophagy markers and caspase-3 immunoreactive cells. Furthermore, our results revealed that CARD was able to attenuate damage via activation of Nrf2 through the p62-dependent degradation of testicular anti-Kelch-like ECH-associated protein-1 (Keap-1). In conclusion, this study suggests that CARD provides protection against diabetic stress-mediated testicular damage. The use of CARD with conventional anti-diabetic therapy was associated with improved efficacy compared with conventional therapy alone.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.163-168
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• 2005

The aim of this study was to investigate the changes in insulin-like growth factor-I (IGF-I) and growth hormone (GH) in serum, the quantitation of spermato-genesis and the comparable relationships among these measurements during pubertal period in New Zealand White male rabbits. To investigate the age-related testicular changes in DNA contents of spermatogenic cells, the fine-needle testicular biopsies from males aged 10 to 28 wks were evaluated by flow cytometry(FCM). Body weight increased significantly between the ages of 12 and 20 wks (P<0.05) and reached 3.4 kg at 28 wks of age. The highest serum IGF-I level (451.3ng/mL) was observed at 20wks of age (P<0.05) and thereafter remained stable at low levels. Serum GH level at 18 wks of age was 183.3 pg/mL which was significantly higher compared to the other ages (P<0.05), and the rising time in serum GH tend to be somewhat earlier than that of IGF-I. The relative percentage of It-cells in testicular cell compartments was $48.2\%$ at the age of 18 wks which significantly increased than those of 16-wk-old (P<0.05) and thereafter increased with the advance of age to $68\%$. The percentage of 2C-cells in testis was $26.8\%$ at 18 wks of age which was significantly lower than $54.3\%$ at 16 wks old (P<0.05). The percentage of 4C-cells was constantly maintained $2\~6\%$ except the $9.9\%$ at 18 wks of age. In conclusion, the results suggest that the puberty onset occurred at about the 18 wks of age and that the IGF-I and GH in serum during the pubertal period showed the age/growth-specific changes and these changes might be related to the spermatogenesis. The DNA FCM combined with fine-needle testicular biopsy could offer a very sensitive method to monitor the quantitative spermatogenic events related to the puberty onset.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.305-315
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• 2001

In this study, the production of transgenic embryo was attempted by microinjection or round spermatid cultured with foreign DNA. At first, the expression of haploid spermatids specific gene, mTP1 in mouse and hPrm2 in hamster spermatids were investigated by RT-PCR method in testes of young mice and hamster testis. The specific gene expression first appeared at 18 days post partum (dpp) in mice spermatid and 20 dpp in hamster spermatid. Therefore, the round spermatids isolated from 17 dpp mice and 19 dpp hamster were used for the introduction of foreign EGFP gene into haploid round spermatids. For the introduction of EGFP gene haploid round spermatids suspended in medium including EGFP gene were treated with a different electric field strength at 0.11, 0.18 and 0.44 ㎸/cm. After electrical stimulation, viability of testicular sperm cells and 67.6%, 66.4% and 49.9%, in mice and 62.6%, 57.9% and 27% in hamster, respectively. These values were significantly lower than those of non-treated control groups 80.5% in mouse and 69.1% in hamster After 72 hrs culture, the highest expression rate of EGFP gene, 28.5% in mice and 32.1% in hamster were obtained from tile spermatogenic cells electroporated by the field strength or 0.18 ㎸/cm. Then, the ability of fertilization and embryonic development of haploid spermatids transfected with foreign EGFP gene were estimated by the microinjection of spermatids into hamster oocytes. The Irate pronuclear formation rate (77.5%) was lower than non-treated control (80%), and the cleavage rate of the treated group (58.8%) was lower than control (65%). To prove the foreign EGFP integration in hamster embryos, 2-cell stage hamster embryos were subjected to the observation under the fluorescence microscope, and the PCR analysis. As a result, about 44% of 2-cell embryos were showed the integration of EFGP gene into their genome. Therefore, These results suggest the possibility to produce transgenic hamsters by microinjection of haploid spermatid transfected with foreign DNA.

• The Korean Journal of Malacology
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• v.32 no.3
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• pp.197-202
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• 2016

This research reports the intersexuality of bivalves, such as Crassostrea gigas, Mytilus galloprovincialis, Rupiditapes philippinarum, Gomphina veneriformis and Barnea davidi discovered during the process of investigating the ecological health status of coastal waters of Korea. In intersex ovaries, the opposite germ cells were observed either individually or in groups in the interfollicular space and inside the oogenic follicle. Oocytes in the intersex testis were at the previtellogenic or initial vitellogenic stage. They were either scattered individually or in groups in the interfollicular space and inside the spermatogenic follicle. The intersexuality in C. gigas was 10.4% (n = 19/183), while female (12.2%, n = 6/49) exhibited a higher proportion than male (9.7%, n = 13/134). The intersexuality in M. galloprovincialis was 31.7% (n = 19/60), while female (36.4%, n = 12/33) exhibited a higher proportion than male (25.9%, n = 7/27). The intersexuality in R. philippinarum was 11.2% (n = 11/98), while male (16.7%, n = 7/42) exhibited a higher proportion than female (7.1%, n = 4/56). The intersexuality in G. veneriformis was 28% (n=30/107), while male (31.5%, n=17/54) exhibited a higher proportion than female (24.5%, n=13/53). The intersexuality in B. davidi was 18.4% (n = 7/38), while female (35.7%, n =5 /14) exhibited a higher proportion than male (8.3%, n = 2/24).

• Development and Reproduction
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• v.11 no.1
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• pp.31-41
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• 2007

The spermiogenesis of Crocidura shantungensis were studied by electron microscope. All process of spermiogenesis was divided into 11 phases 15 steps, based on the morphological features of the nucleus and cell organelles in cytoplasm of spermatids. The spermatids in Golgi and cap phases were a spherical shape. On the other hand, at the early acrosomal phase they changed into an oval shape, and the tail was created in this phase. In maturation phase, the shapes of spermatid head were thin and longish. Until step 7 the direction of spermatids head turned toward the lumen of the seminiferous tubule. From step 8 to step 15 their heads turned toward the basal lamina. In step 12, the nucleus and acrosome shown maximal elongation. From Step 13 the nucleus of spermatids became flat, simultaneously with flat expansion of the acrosome expanded, and the visible whole lengths of spermatids were tend to be shorten. Spermatid heading which arrived to step 14 was taken the final shape. The nucleus was doing the wedge shape, and the nuclear chromatins condensed completely and homogenized. In the spermiation phase, the spermatids were gradually disconnected from the cytoplasm of the Sertoli cell. In this phase, the acrosome of the spermatids were fully shorten and flat, and the spermatozoa completed the process of heading and the tailing. Considering all the results, the spermiogenesis may be useful information to analyze the differentiation of spermatogenic cells.

• Development and Reproduction
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• v.11 no.2
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• pp.97-104
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• 2007

This study describes the spermatogenesis and sperm ultrastructure of the spiny top shell, Batillus cornutus using light and electron microscopy. The spiny top shells were collected by divers in the coastal water of Wandogun, Cheollanamdo, Korea(N $34^{\circ}13'$, E $126^{\circ}47'$) at May 2003. Spiny top shells of $60.0{\sim}69.9\;mm$ in shell height were used in this study. The testis comprises many spermatogenic follicles which contains germ cells in different developmental stages. The primary spermatocytes in the pachytene stage were characterized by synaptonemal complexes. The early spermatids were characterized by appearance of Golgi complex, increased karyoplasmic electron density and tubular mitochondria. In early spermatid the mass of proacrosomal granules consists of numerous heterogeneous granules with high electron density. From the mid-stage of spermiogenesis the well-developed mitochondria aggregate posterior to the nucleus, and surround the proximal and distal centrioles. In this stage, proacrosomal granules are condensed and form a acrosome with thin envelope. During the late spermiogenesis, the acrosome begins to elongate and then became conical. The sperm consists of head, mid-piece and tail. The head comprises a round nucleus and a conical acrosome. Acrosomal rod of microfibrous is observed between nucleus and acrosome. Five mitochondria observed in mid-piece. And tail has the typical "9+2" microtubular system originates from the centrioles.

• Development and Reproduction
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• v.14 no.2
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• pp.99-105
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• 2010

The seminiferous epithelium, with its division into 12 spermatogenic stages in the mouse, is a very complex tissue. Glutathione peroxidase (GPx) is a representative antioxidant enzyme that is capable of reducing organic hydroperoxides to their corresponding hydroxyl compounds utilizing glutathione and is related to the mammalian spermatogenesis. In this study, a real-time PCR was performed in the stage-specific seminiferous tubules of mouse testes excised by a laser capture microdissection (LCM) in order to quantitate the expression levels of a series of GPx genes including cytosolic GPx (cGPx), gastrointestinal GPx (GI-GPx), plasma GPx (pGPx), and phospholipid hydroperoxide GPx (PHGPx). Frozen sections (10 ${\mu}m$) were obtained from normal adult mouse testes. LCM was used to capture all the cells that were grouped into stages I-V, VII-VIII, and IX-XI in cross-sections of seminiferous tubules. The expression level of PHGPx mRNA was remarkably higher than those of other GPx mRNAs in mouse testes. During spermatogenesis, the expressions of GI-GPx, pGPx, and PHGPx mRNAs were highest on stages VII-VIII, began to decrease after stage XI, and showed a lowest level on stage I-V. However, the expressions of cGPx mRNA were highest on stages VII-VIII, and showed a lowest level on stage XI-XI. These findings indicate that GPx genes are expressed differentially on mouse spermatogenesis and also LCM can be an useful tool in cellular quantitative analysis of testes.