• Asian-Australasian Journal of Animal Sciences
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• v.4 no.2
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• pp.151-156
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• 1991

Bovine in vitro fertilization experiment was carried out using ovary-derived follicular oocytes and frozen-thawed spermatozoa to determine the optimal preincubation time of spermatozoa and the insemination time for successful in vitro fertilization rate. The possibility of parthenogenetic cell division of unfertilized oocytes during culture without spermatozoa was also examined. There was no significant (p>0.05) difference in percent ratio of embryos developed to blastocyst stage between 0 and 3 h preincubation times of spermatozoa, showing a tendency to have higher percentage for 0 h of preincubation time. The 6 h insemination time seemed to be better for producing higher percentage of ova cleavage compared with those of 1 and 3 h treatments. Approximately 10% of unfertilized oocytes divided into 2 to 4-cell stage, and some of them cleaved to 5 up to 8-cells. The results obtained from this study suggested that 0 h of sperm preincubation time and 6 h of insemination time would be suitable for producing better in vitro fertilization rate of bovine oocytes. It is also likely that unfertilized bovine oocytes probably cleave to some cell stages with irregular divisions of the cells. On the one hand, considerable variation was also found in spermatozoa function among individual bulls.

• Development and Reproduction
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• v.19 no.4
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• pp.197-207
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• 2015

This study investigated structure and function of the reproductive system in Aplysia kurodai by means of anatomical, histological, and histochemical observation. Reproductive system of this species is consisted of ovotestis, small hermaphroditic duct, ampulla, accessory genital mass and large hermaphroditic duct. The ovotestis is composed of a large number of follicles, and both oocytes and spermatocytes matured in the same follicle. The small hermaphroditic duct is a single tube and contains a swelling, the ampulla, which functions as a storage organ for endogenous sperm and an oviduct. The accessory genital mass is connected to both the small and large hermaphroditic duct, and consisted of three glands: albumen, membrane (winding) and mucus gland. The albumen gland is consisted of granular cells producing basophilic and neutral mucopolysaccharides. The membrane and mucus gland are consisted of granular cells producing acidophilc and sulfated mucopolysaccharides. The large hermaphroditic duct is a single tubular gonoduct linking the accessory genital mass to the common genital aperture but is consisted of two parallel compartments. Internally, these two compartments are incompletely divided by internal septum or fold, which are called as the red hemiduct and white hemiduct, respectively. The red hemiduct functions as an oviduct and the white hemiduct functions as a copulatory duct. The reproductive system of A. kurodai is externally comprised a single tube, i.e., monaulic type. However, internal structure of duct is incompletely divided into oviduct and copulatory duct, i.e., the oodiaulic type.

• Korean Journal of Veterinary Research
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• v.23 no.2
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• pp.137-148
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• 1983

This study was made for the better information of the male reproductive system on the meat-type drake, Cherry Belly X White Golden. The epithelium of ductules of epididymal region and deferent duct were observed histologically and histochemically with the progress of their development. India-ink absorbability on the luminal epithelium was also investigated after the administration of India-ink. The results are as follows; 1. Rete testis and various round ductules in immature form appeared in epididymis within 6 weeks after hatching, and simple cuboidal and simple columnar epithelium were found in the epithelia of the ductules within 8 weeks after hatching. Larger ductules were found on epididymal surface which was in the developing stage near to the immature efferent ductule. From 10th to 20th week, various ductules appeared in epididymis, and developing form of efferent ductules were much more increased on epididymal surface. The luminal epithelium of the ductules were composed of ciliated simple columnar and pseudostratified ciliated columnar cells. At the same time, deferent duct appeared. From the 21th week, various ductules in epididymis became abruptly matured. Lumen of rete testis was lined by simple squamous or simple cuboidal epithelium, and that of efferent ductules, having many folds and being larger than any others were lined by pseudostratified ciliated columnar epithelium in which ciliated columnar cells, non-ciliated cells(clear cells) and basal cells were noted. Connecting tubules of star shaped lumen were composed of pseudostratified ciliated columnar epithelium in which ciliated columnar cells, nonciliated cells, and basal cells were observed. The luminal surface of epididymal ducts was smooth and has thick pseudostratified columnar epithelium which was composed of high columnar cells and basal cells. From 26th week after hatching, sperm pooling was started in various ductules. 2. From 4th to 10th week, simple cuboidal epithelium of deferent duct transformed to simple columnar epithelium with the progress of aging. At the basement of epithelium, clear round cells were noted. From 12th to 20th week, high columnar cells with enlongated nucleus were noted on the luminal border of deferent ducts, forming folds of pseuclostratified columnar epithelium. From 20th week, the deferent duct started to have septa in it's lumen and composed mainly of pseudostratified columnar epithelium, and round cells disappeared. From 20th week, the lumen diameter of deferent duct became wider with the progress of aging, but there was no difference among the values of lumen diameter in upper, middle, and lower part of deferent ducts. At 26th week, the pooling period of sperms in deferent ducts, the lumen diameter became rapidly widen, especially in the lower part of deferent ducts. Thickness of muscular layer of ductus deferens showed gradual growth within 24 weeks but did abrupt thickening from 26th week. 3. Saliva resistant PAS granules were dotted on the top of nucleus in efferent ductules epithelium but the amount of the granules were little in the connecting ductules's epithelium. The granules reactive to acid phosphatase were abundant in the some epithelial cells of efferent ductules and connecting ductules, especially above the nucleus of cells. The granules reactive to alkaline phosphatase were noted on the luminal border of efferent ductules. Parts of free border of efferent ductules and middle portion of deferent ducts were stained slightly by alcian blue technique. India ink granules were found mainly in the epithelium of efferent ductules but were few in that of connecting ductules.

• The Korean Journal of Pesticide Science
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• v.5 no.4
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• pp.11-19
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• 2001

This review discusses the effects and roles of insect hormones and insect growth regulators (IGRs) on vitellogenesis in adult insects. Insect vitellogenesis is regulated by hormones such as juvenile hormone (JH), ecdysteroids, and neurosecretory hormones (ovaryecdysteroidogenic hormone : OEH) released by neurosecretory cells, diet, and other elements(male specific protein of sperm fluid). In the fat bodies, the vitellogenins are synthesized by the stimulation of JH released by corpus allatum (CA) and ecdysteroids produced by follicle cells with the ovary in most insects. Furthermore, vitellogenins are released into the hemolymph, transported to the ovarioles by carrier protein, and incorporated into oocytes for the developing ovary. Of IGRs, juvenile hormone and its mimics such as methoprene and pyriproxifen appear to have pharmacological effects such as membrane lysis, destruction of salivary grand and midgut epithlial cells, fat body cells, and ovarian tissue, and also anti-juvenile hormone such as precocenes I and II appear to have specific cytotoxicity such as inhibition of corpus allatum and oocytes development. These results suggest that IGRs may be useful as agents for integrated pest management.

• 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.

• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.4
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• pp.424-430
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• 2002

Vas deferens is a long tube and could be divided into four regions as its morphological characters: a short and slender proximal region, a convoluted region, a straight and more thick distal region, and an ejaculatory duct, It is connected with the posterior outer horns of the testis and runs to the gonopores opened on the coxa of the fifth walking legs, The proximal region consists almost entirely of simple cuboidal epithelial cells, $12\~28{\mu}$m in height, surrounded by a thin basal membrane. A small aggregation of high cuboidal epithelium is obsened from one side of the proximal region. The convoluted and distal region is composed of two kinds of epithelial cells; high cuboidal epithelial cells, $40\~120{\mu}m$ in height located in dorsal portion and simple cuboidal epithelial cells of $12\~28{\mu}$m in height located in ventral and lateral portion. The ejaculatory duct is surrounded with two kinds of muscle layers, inner longitudinal and outer circular muscle fibers, The lumen is lined with high and simple cuboidal epithelium in almost equal proportions. The proximal region contains mature sperm and basophilic substances. The eosinophilic substances appeared newly going toward the convoluted region and are laid along simple cuboidal epithelial cell layer. Tube-like content (spermatophore) of the vas deferens is ejaculated from a pair of genital pores at mating. Two masses of the content fuse together side by side and are usually deposited on the female thorax between the second and fourth or fifth pereiopods, The spermatophore formed measures $2.7\~4.0$ mm in length and $1.5\~2.7$ mm in width.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.333-340
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• 2003

Objective: This study was performed to examine the maturation and the development to the blastocyst stage of immature oocytes collected from patients with high risk of ovarian hyperstimulation syndrome (OHSS). Materials and Methods: Cumulus-oocyte complexes (COCs) were collected following only HCGpriming for non stimulated IVF-ET cycles of the patients. At the time of oocyte collection, COCs were classified into three groups in accordance with their appearance (Group I: oocytes with dispersed cumulus cells; Group II: oocytes with compacted cumulus cells; Group III: oocytes with sparse cumulus cells). The in vitro maturation and blastocyst development rates of the COCs were compared among these groups. From August 2001 to June 2002, 48 IVM/IVF-ET cycles from 42 patients (mean age: $32.4{\pm}3.8$ years) were performed. To prevent the occurrence of OHSS, the patients were primed with 10, 000 IU HCG alone 36 h before oocyte collection without gonadotropin stimulation. Oocytes were aspirated on cycle days from 7 to 13. The normal COCs were classified into three groups according to their appearance. The aspirated immature oocytes were cultured in YS maturation medium containing 30% (v/v) human follicular fluid (HFF), 1 IU/ml FSH, 10 IU/ml HCG and 10 ng/ml rhEGF. Fertilization was induced by intracytoplasmic sperm injection (ICSI). All zygotes were co-cultured with cumulus cells in $10{\mu}l$ YS medium containing 10% HFF until day 7 after oocyte collection. Blastocyst transfer was performed on day 5 after ICSI. Results: Th e mean number of oocytes cultured in the IVM/IVF cycles was $24.7{\pm}10.6$. Of 1185 COCs, those assigned to Group I, II and III were 470 (39.7%), 414 (35.0%) and 301 (25.4%), respectively. The maturation rate (94.5%, 444/470, p<0.05) in Group I was significantly higher than those of Group II (62.8%, 260/414) and Group III (73.1%, 220/301). Especially, 30.9% of COCs in Group I (145/470) was matured on the day of oocyte aspiration. There were no differences in the rates of fertilization and cleavage among the three groups. The development rate to the blastocyst stage in Group I (54.6%, 206/377, p<0.05) was also significantly higher than those in Group II (33.0%, 68/206) and Group III (30.1%, 52/173). Twenty-four clinical pregnancies (50.0%) was obtained and 22 pregnancies (45.8%) are ongoing. Implantation rate in the present study was 24.6%. Conclusion: These results suggest that there is a positive correlation between the appearance of COCs and the developmental competence of the immature oocytes in non stimulated IVM/IVF cycles.

• The World Journal of Men's Health
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• v.41 no.1
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• pp.215-226
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• 2023

Purpose To clarify (phospho-) glycogen synthase kinase-3 (GSK3) isoform variants in the germline and soma of human testes and spermatozoa. Materials and Methods GSK3 isoform variants in normospermatogenic and Sertoli cell-only (SCO) testicular biopsies and spermatozoa were examined. In normospermatogenic testes, GSK3α and GSK3β variants 1 and 2 different in low complexity region (LCR) were expressed and their levels were decreased in SCO testes. GSK3β variant 3 was only expressed in SCO testes. GSK3β as well as GSK3α, the dominant isoforms in testes were decreased in SCO testes. In normospermatogenic testes, GSK3β were found in spermatogonia and markedly decreased in meiotic germ cells in which GSK3α was dominant. p-GSK3α/β were marginal in spermatogonia and early spermatocytes. In SCO testes, GSK3α/β immunoreactivity in seminiferous epithelia was weaker than those of normospermatogenic testes whereas p-GSK3α/β(Ser) immunoreactivity was visibly increased in Sertoli cells. GSK3α was dominant in ejaculated spermatozoa in which GSK3α and p-GSK3α(Ser) were found in the head, midpiece, and tail. In acrosome-reacted spermatozoa, GSK3α was found in the equatorial region of head, midpiece, and tail, and p-GSK3α(Ser) was only found in midpiece. During sperm capacitation, p-GSK3α(Ser) was significantly increased together with phosphotyrosine proteins and motility. In human male germ cells, GSK3 isoforms different in LCRs switch from GSK3β to GSK3α during meiotic entry, suggesting the isoform-specific roles of GSK3α and GSK3β in meiosis and stemness or proliferation of spermatogonia, respectively. In dormant Sertoli cells of SCO testes kinase activity of GSK3 might be downregulated via inhibitory phosphorylation. In spermatozoa, inhibitory phosphorylation of GSK3α might be coupled with activation of motility during capacitation.

• Journal of Embryo Transfer
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• v.30 no.2
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• pp.109-114
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• 2015

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.358A>T) of cluster-of-differentiation antigen 9 (CD9) gene reported to be significant association with MOT. Also, CD9 gene was expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as candidate gene for boar semen. This study was conducted to evaluate the pig SNP (g.358A>T) of CD9 gene as a positional controlling for semen parameters of post-thawed boar semen. To results, the g.358A>T SNP of the CD9 gene was significantly associated with the traits such as MOT, curve linear velocity, straight line velocity, average path velocity and amplitude of lateral head displacement. Particularly, the g.358A>T SNP significantly has the highest association with MOT and animals with AA genotype (p<0.001). Therefore, we suggest that the g.358A>T in the intron 6 region of the porcine CD9 may be used as a molecular marker for Duroc boar Post-thawed semen quality, although its functional effect was not defined yet.