• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.413-431
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• 1992

Gonadal development, fertilization and egg development in the maternal body and reproductive cycle of ovoviviparous rockfish, Sebastes inermis, were investigated histologically. Gonadosomatic index(GSI) of male and female were increased from September and reached maximum values in December. In the male, GSI decreased from January, but in the female maintained high values till February and decreased from March. Hepatosomatic index(HSI) was related to GSI conversely. In both sex, HSI increased from February and reached maximum in August as the gonad were degenerating and resting, and began to decrease from September as gonad were glowing. This ovoviviparous rockfish copulates in December. Fertilization with sperms maintained between ovulated oocytes in the ovary occurs in January mainly. Egg development in the ovarian cavity and discharging of hatched preiarva occurs from January to February. The reproductive cycle includes the successive stages: Growing(September), Mature (October-November), Ripe and Fertilization(Decembr-Janua), Egg development and Discharging of hatched larva(January-February), Degeneration and Resting(February-August). According to the frequency distribution of egg diameter and histological observation, the ovoviviparous rockfish discharged the prelarva at a time in a spawning season. The sexual maturation is first attained at 2 ages. All females and males reaches first maturity at body length of 17.1cm and 15.1cm respectively. The mean number of the embryos increased with the increase of the total length of female.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.23-29
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• 2000

Objective: Zona pellucida (ZP) has been thought to be the barrier of egg to sperm penetration before and after fertilization. The phenomenon of ZP hardening has been considered as a post-fertilization event until now, and it is generally accepted that it is caused by the secretory products of cortical granules released during the cortical reaction. Hardening of ZP could occur "spontaneously" in mammalian oocytes in standard culture conditions, and that it is probably not a consequence of cortical reaction. The purpose of our study was to investigate the effect of human amniotic fluid (HAF) on nuclear maturation (NM) and fertilization ability of mouse immature oocytes. Methods: HAF was obtained from patients undergoing amniocentesis at $16{\sim}20$ weeks of gestation. HAF from five to ten patients was centrifuged and the supernatants was pooled. Cumulusenclosed mouse immature oocytes were incubated in the medium containing HAF, and examined to confirm NM and fertilization. Female ICR mice (about 3 weeks old) were stimulated with 7.5 IU PMSG. Immature oocytes were isolated at $48{\sim}52$ hrs post PMSG injection and cultured in TCM-199 supplemented with 20% HAF for 18 hrs. FBS was used as a control for the examination. Matured oocytes (MII) were fertilized with sperms collected from the epididymis of male mice (over 10 weeks old). Fertilization was in conducted T6 medium containing 15 mg/ml BSA, and confirmed at 6 hrs post-insemination. Fertilization rate was assessed in zona-intact or zona-free oocytes (denuded by trypsin). Evaluation of NM and fertilization was carried out by rapid staining method. ZP hardening was evaluated by incubating cumulus cell-free mature oocytes in 0.001% chymotrypsin at $37^{\circ}C$ for 10 min. Results: There was no significant difference between the effects of HAF (86.6%) and FBS (87.7%) supplements on NM of immature oocytes. When maturation medium was supplemented with HAF, total fertilization rates (7%) were significantly lower (p<0.01) than that of FBS (85.1%). In HAF group, fertilization rate was increased (p<0.01) in zona-free oocytes (7% versus 100%). The resistance of mouse oocyte ZP to digestion by chymotrypsin after maturation in vitro was significantly higher (p<0.01) in HAF group (86.7%) than in FBS (6.7%). To culture oocytes in FBS were very effective in preventing ZP hardening. However cultured oocytes in HAF showed high rate of ZP hardening (p<0.01). Conclusions: These results suggest that HAF can be used as a supplement for the NM of mouse immature oocytes in vitro. However, HAF induces spontaneous hardening of ZP of mouse immaure oocytes during maturation in vitro.

• Journal of Life Science
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• v.27 no.10
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• pp.1097-1103
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• 2017

For the artificial induction of the sexual maturation of Anguilla japonica, salmon pituitary extract (SPE) is continuously injected into females, and the eggs obtained from artificial sexual maturation are artificially fertilized with sperms and hatched. However, repeated injection of SPE in the abdominal cavity causes tremendous stress in females, which may prevent their complete sexual maturation and reduce the immune system function, ultimately resulting in death. In addition, the poor quality of the ovulated eggs can reduce the hatching and survival rate of larvae. In the present study, sexual maturation of females was induced by inserting an osmotic (OS) pump containing hormone analogs known to effectively induce sexual maturation into the abdominal cavity of female eels, and the effect of the OS pump on the induced sexual maturation was investigated. Our study results showed that the gonadosomatic index (GSI) was significantly higher in the fish subjected to SPE injection than those subjected to human chorionic gonadotropin (hCG), gonadotropin-releasing hormone analog (GnRHa), and methyl testosterone (MT) injections, either separately or in combination. In addition, a histological analysis showed that the oocytes in the SPE OS pump groups were more mature (entered the nuclear shift stage) than those in the other groups. These results suggest that an osmotic pump containing hormone analogs can be used to induce sexual maturation in female A. japonica artificially.

• The Korean Journal of Malacology
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• v.26 no.3
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• pp.235-244
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• 2010

Ultrastructural characteristics of the testis and spermatogenesis of Crassostrea gigas were investigated by Transmission and Scanning Electron microscope observations. The testis is a diffuse organ consisting of branching acini containing differentiating germ cells in a variety of stages. The acinus is surrounded by an intermitent layer of myoepithelial cells andis divided into subcompartments that are partially separated by pleomorphic accessory cells which remain in close contact with germ cells until late stages of development. these accessory cells contain a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in the supplying of the nutrients for germ cell development, while any phenomena associated with phagocytosis of undischarged, residual sperms by lysosomes could be find in the cytoplasm of the accessory cells. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves. Mature spermatozoa consist of broad, cap-shaped acrosomal vesicle, subacrosomal material (containing axial rod embedded in a granular matrix), a oval nucleus showing deeply invaginated anteriorly, two triplet substructure centrioles surrounded by four spherical mitochondria, and satelite fibres appear to the distal centriole and plasma membrane. Spermatozoa of C. gigas resemble to those of other investigated ostreids. In particular, the anterior region of the acrosomal vesicle is transversely banded. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm morphology in the resolution of taxonomic relationships within the Ostreidea. The spermatozoon is approximately $42-47{\mu}m$ in length including an oval sperm nucleus (about $0.91{\mu}m$ in length), an acrosome (about $0.42{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9 + 2 structure. These morphological charateristics of acrosomal vesicle belong to the family Ostreidae in the subclass Pteriomorphia.

• Journal of Aquaculture
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• v.21 no.4
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• pp.331-338
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• 2008

Monthly changes in the gonadosomatic index (GSI) and hepatosomatic index (HSI) of wild river puffer Takifugu obscurus, and water quality environment in spawning area during breeding season were investigated from March 1995 to February 1996. Monthly changes in GSI and HSI of T. obscurus, that was cultured in low salinity, were calculated. The external morphology of the gonads, germ cell differentiation during gametogenesis and the reproductive cycle with the gonad developmental phases were investigated by histological analysis. The optimum water quality environment in Ganggyung, Choongcheongnam-do, where is spawning ground of wild T. obscurus, was $15-20^{\circ}C$ (water temperature) and 0 psu (salinity). Monthly changes in the GSI in females and males reached a maximum in May, and then rapidly decreased. Therefore, it is assumed that in the natural condition the spawning period of wild T. obscurus is May to June. In females and males, it showed a negative correlationship between the GSI and HSI. The external morphology of the gonads in female and male T. obscurus, that was cultured in low salinity, is composed of a pair of saccular structure. Based on monthly changes in the GSI, it is assumed that in female T. obscurus, that was cultured in low salinity, spawn from March through May. Therefore, it showed a negative correlationship between changes in the GSI and HSI. On the whole, in females and males, it showed a similar pattern between wild and cultured T. obscurus. The reproductive cycle with the gonad developmental phases can be classified into successive five stages in females: the early growing stage, late growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In males, that can be divided into successive four stages: the growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In case of wild T. obscurus, the spawning period has once a year, however, those cultured in the high water temperature ($20-27^{\circ}C$) - low salinity (under 3.3 psu) condition have reproductive characteristics having possibilities of discharge of eggs and sperms year-round as a multiple spawner.

• The Korean Journal of Malacology
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• v.30 no.4
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• pp.311-319
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• 2014

Early sexual maturation through temperature stimulation was induced in female and male of yezo scallop. Gonadosomatic index (GSI) in female showed $9.12{\pm}2.9$ in January, $14.89{\pm}2.9$ in February and $21.3{\pm}1.4$ in March in experiment I. GSI in experiment I showed a significant increase (P < 0.05) and in experiments II and III were not show significant variations (P > 0.05). It also showed significant between the control and the experiments I, II, and III in February (P < 0.05) measurements. Experiment I has showed good results in sexual maturation and spawning when compared with other experiments II and III and the control. Histological observation showed that ovary condition was in a growing stage in all the experiments I, II, and III. In February, ovary condition through histological observation was a late mature stage in all the experiments I, II, and III except the control of a growing stage. GSI and gonad weight were $4.4{\pm}0.88$ and 2.8 g, respectively in November whereas it was $15.1{\pm}2.8$, and 11.7 g, respectively in January and $21.7{\pm}5.4$, and 19.4 g, respectively in February after rearing at a water bath of $12^{\circ}C$ depending on the condition of experiment I. It was possible early releasing of eggs and sperms of yezo scallop in February instead of the middle of April to the end of May being spawning period. Fertilized eggs have become a gastrula stage through a spiral cleavage and then become a trochophore larvae after 36 hours. After 10 days, D-shaped larvae have changed into an umbo stage larvae and attached to juveniles in the post larvae after 20-23 days.

• Development and Reproduction
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• v.14 no.4
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• pp.269-279
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• 2010

Some characteristics of germ cell differntiations and the function of accessory cells during spermatogenesis, and mature sperm ultrastructure in male Protothaca (N.) jedoensis were investigated by transmission electron microscope observations. The morphology of the spermatozoa of this species has a primitive type and is similar to those of other species in the subclass Heterodonta. Accessory cells, which are connected to adjacent germ cells, are involved in the supplying of the nutrients for germ cell development. The morphologies of the sperm nucleus and the acrosome of this species are the cylindrical type and cap shape, respectively. Spermatozoa are approximately $46{\sim}50{\mu}m$ in length including a long sperm nucleus (about $2.44{\mu}m$ in length), an acrosome (about $0.45{\mu}m$ in length), and tail flagellum (about $42{\sim}46{\mu}m$). The axoneme of the sperm tail shows a 9+2 structure. As some characteristics of the acrosomal vesicle structures, the basal and lateral parts of basal rings show electron opaque part (region), while the anterior apex part of the acrosomal vesicle shows electron lucent part (region). These characteristics of the acrosomal vesicle were found in the family Veneridae and other several families in the subclass Heterodonta. These common characteristics of the acrosomal vesicle in the subclass Heterodonta can be used for phylogenetic and systematic analysis as a taxonomic key or a significant tool. The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appear in most species in the family Veneridae and other families in the subclass Heterodonta. However, exceptionally, only three species in Veneridae of the subclass Heterodonta contain 5 mitochondria. The number of mitochondria in the sperm midpiece can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or an important tool.