• Journal of Embryo Transfer
• /
• v.13 no.1
• /
• pp.1-9
• /
• 1998

In order to improve the cryopreservation by vitrification or slow freezing of nuclear transplant rabbit embryos, the effects of factors affecting embryo cryopreservation such as cryoprotectants, equilibration, cooling rate and post-thaw dilution on post-thaw survial and development were determined using intact embryos of morular stage. And the post-thaw development of nuclear transplanted embryos cryopreserved under the optimal conditions examined was compared between vitrification and slow freezing. The cryoprotectant solution used was ethyleneglycol-ficoll-sucrose (EFS) or ethyleneglycol-poly-vinylpyrrolidone-galactose- I (EPG- I ) for vitrification, and EPG- II for slow freezing. To examine the viability of frozen-thawed embryos, the nuclear transplanted embryos were co-cultured in TCM-199 plus 10% FBS with bovine oviduct epithelial cells(BOEC) for 24 hrs and the intact morulae were co-cultured with BOEC for 5 days and 3 days to hatching blastocyst stage in 39 ˚C 5% $CO_2$ incubator. The results obtained were as follows: Following vitrification with EFS, the post-thaw development of rabbit morulae to hatching blastocyst was significantly(P<0.05) higher in compacted stage(82.4%) than in early morular stage(60.0%). The post-thaw development of compacted morulae to hatching blastocyst was similarly high in vitrification with EFS(82.4%), EPG- I (85.0%) and in slow freezing with EPG- II (83.3%). Following vitrification with EPG- I, the post-thaw development of intact rabbit morulae to hatching blastocyst was similar as 78.0% and 85.0% in 1-step and 2-step post-thaw dilution, respectively. The post-thaw development of nuclear transplanted rabbit embryos of compacted morulae stage to hatching blastocyst was similarly 43.6% and 40.0% in vitrification with EPG- Iand slow freezing with EPG- II, respectively. These results indicated that the rabbit nuclear transplant and intact embryos of morulae stage could be well cryopreserved with either vitrification or slow freezing procedure.

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.1
• /
• pp.35-42
• /
• 1997

The present study was designed to define the role of prostaglandin in the development and hatching of mouse embryo. The effects of indomethacin, an inhibitor of prostaglandin synthesis, on the development and hatching of morula and blastocyst were examined. In early morula stage, embryos were degenerated significantly at 100 ${\mu}M$ and 200 ${\mu}M$ indomethacin. However, the viability of embryos was not influenced by concentration in any other embryonic stages. In all embryonic stages, the hatching was suppressed with concentration dependent manner, but expansion was not suppressed. Particularly, in 84h embryos post hCG injection, the hatching was suppressed significantly compared with post hCG 72h or 96h embryos. When embryos were treated with 100 ${\mu}M$ indomethacin for a specific time (12h) in according to the development stage, the hatching was suppressed all groups. These suppressional effect was decreased as embryonic development stage was progressed. However, the expansion was not affected in all treatment group. This study suggests that hatching-related metabolic substances are synthesized from morula stage and intraembryonic signaling mediated prostaglandin was important for development and hatching of mouse embryo.

• Clinical and Experimental Reproductive Medicine
• /
• v.32 no.4
• /
• pp.337-346
• /
• 2005

Objective: The oligosaccharide moieties of glycoproteins and proteoglycans have a vital function in blastocyst differentiation. Concanavalin (ConA), a lectin, is known to bind on the preimplantation embryos, especially on blastocyst. In this study, we investigated whether ConA can modulate the trophoblast development and about the regulating mediator. Also, we investigated whether expansion is enough for hatching procession of the mouse blastocyst. Method: Embryos were collected at 72 h post hCG injection and chemicals were treated after 24 h (96 hr post hCG injection). ConA or calcium ionophore A23187 were exposed to blastocyst and than analysis the developmental process for 48 hr. Intracellular free-$Ca^{2+}$ concentration in trophectoderm was measured with confocal laser microscope after exposing to ConA or calcium ionophore A23187. ConA-pretreated blastocyst exposed to the calcium ionophore A23187 and then analyzed the developmental process. Otherwise ouabain was treated to the blastocyst to block the $Na^+/K^+$-ATPase activity. Results: In contrast to the control blastocyst, the ConA-exposed blastocysts developed beyond the expansion stage with significantly high rate (90.4%) at 12 h post administration. ConA induced an increase the intracellular $Ca^{2+}$ concentration in trophectoderm. Calcium ionophore A23187 also stimulated expansion of blastocyst. Most of the control blastocysts developed to the hatching stage at 144 h post hCG injection. However, strongly 65% of the ConA-exposed embryos were arrested at expanded stage at same time point. The developmental progression rates to hatching stage of both ConA- and calcium ionophore A23187-expose blastocysts were significantly lower than that of the control. However ConA-pretreated embryos developed to the hatching stage like control embryos. Ouabain showed a tendency to delayed the progress to expansion stage but did not inhibit the development to the hatching stage. Conclusion: ConA-mediated expansion is the result of the increase of intracellular free-calcium in blastocyst stage embryo. It is suspected that expansion of the blasocyst is a essential indirect factor in hatching and the calcium may triggering the cellular mechanisms for the both expansion and hatching progression.

• Korean Journal of Ichthyology
• /
• v.36 no.1
• /
• pp.20-29
• /
• 2024

Hemitripterus villosus, a promising aquaculture fish species, is facing declining stocks. This study aims to provide normative standards for skeletal development to address persistent skeletal deformities in farmed fish. Specimens utilized in the study underwent artificial insemination with captured fish, and the resulting larvae and fry were preserved in a formalin solution. The skeletal ossification process commenced immediately after hatching, affecting the parasphenoid, premaxillary, maxillary, and dentary structures at an average total length of 13.65±0.71 mm (n=5). By sixty-five days post-hatching, ossification extended to the ethmoid and supraorbital, completing the head's development at an average total length of 21.24±0.50 mm (n=5). Clavicle ossification occurred at seven days post-hatching, corresponding to an average total length of 14.61±0.52 mm (n=5). At forty-four days post-hatching, the ossification of 4 actinosts took place, completing the shoulder girdle, with an average total length of 18.15±0.61 mm (n=5). Vertebral ossification initiated at ten days post-hatching, with an average total length of 14.80±0.65 mm (n=5). By fifty-four days post-hatching, 39 vertebral columns were ossified, reaching an average total length of 18.67±0.54 mm (n=5). Vertebral development was complete at sixty days post-hatching, with an average total length of 20.25±0.45 mm (n=5). This study sheds light on the skeletal development of H. villosus, providing valuable standards and fundamental data for understanding skeletal deformities in this species.

• Development and Reproduction
• /
• v.21 no.1
• /
• pp.63-69
• /
• 2017

On the 15 days after hatching, the larvae was 4.24-5.10 mm (mean $4.66{\pm}2.18mm$) in total length, and the fins of the membrane started to develop into a fan shape and the melanophore was deposited upper the alimentary canal of the abdomen and on the bladder. At 35 days after hatching, the post-larvae formed a branch-shaped melanophore on the head part with a total length of 6.98-12.5 (mean $9.35{\pm}1.71$) mm, formed on the upper and lower parts of the caudal part, formed on the upper and lower parts of the caudal part, and deposited under the head part and abdomen. At 40 days after hatching, the juvenile was 11.3-18.1 (mean $14.9{\pm}1.53$) mm in total length.

• Journal of Environmental Science International
• /
• v.12 no.6
• /
• pp.593-598
• /
• 2003

The effects of bisphenol A on the catalase activities in the development stage of zebrafish were investigated. In this study, the catalase activities for zebrafish fries exposed to bisphenol A of 1${\times}$10$\^$-10 g/$\ell$ during 1 week, 2 week, and 4 week post-hatching were examined. Also, the changes of organs weight and the catalase activities for adult zebrafishes exposed to bisphenol A during 3 weeks were investigated. Catalase activities for zebrafish fries exposed to bisphenol A of 1${\times}$10$\^$-10/ g/$\ell$ during 1 week post-hatching were significantly lower, compared to the control. Somewhat, for zebrafish fries exposed to bisphenol A during 4 week post-hatching, catalase activities were significantly increased. For adult zebrafishes, the effects of bisphenol A were higher for female than male. Specially, catalase activities were significantly increased in the ovary of zebrafishes exposed to bisphenol A during 3 weeks. The ovary weight were increased for zebrafishes exposed to bisphenol A during 3 weeks. Catalase activities were increased in the intestine of female exposed to bisphenol A during 3 weeks. Catalase activities were increased in testis exposed to bisphenol A during 3 weeks but there was no significance. In conclusion, the damages of an endocrine disrupter were higher in the earlier development stage compared with adult. The damages were higher for female exposed to an endocrine disrupter compared with male.

• Development and Reproduction
• /
• v.20 no.1
• /
• pp.31-40
• /
• 2016

This study was conducted in order to examine the egg development in red spotted grouper, Epinephelus akaara and the morphological development of its larvae and juveniles, and to obtain data for taxonomic research. This study was conducted in June 2013, and 50 male and female fish were used for the study. One hundred ${\mu}g/kg$ of LHRHa was injected into the body of the fish for inducing spawning, and the fish were kept in a small-sized fish holder ($2{\times}2{\times}2m$). Eggs were colorless transparent free pelagic eggs, 0.71-0.77 mm large (mean $0.74{\pm}0.02mm$, n=30), and had an oil globule. Hatching started within 27 h after fertilization. Pre-larvae that emerged just after hatching were 2.02-2.17 mm in total length (mean $2.10{\pm}0.11mm$), their mouth and anus were not opened yet, and the whole body was covered with a membrane fin. Post-larvae that emerged 15 days post hatching were 3.88-4.07 mm in total length (mean $3.98{\pm}0.13mm$), and had a ventral fin with two rays and a caudal fin with eight rays. Juveniles that were formed at 55 d post hatching, were 31.9-35.2 mm in total length (mean $33.6{\pm}2.33mm$), with red color deposited over the entire body, and black chromophores deposited in a spotted pattern. The number of fin rays, body color, and shape were the same as that in the adult fish.

• Development and Reproduction
• /
• v.8 no.1
• /
• pp.11-17
• /
• 2004

The larvae and juvenile development of haddock Melanogrammus aeglefinus which is significant commercial fish living north Atlantic Ocean are described here. Larvae were reared in laboratory and sampled periodically for developmental study until 67 days after hatching. An increase in total length(TL) of fish indicated continuous growth, described by the growth expression Y=4.07 $e^{0.037}$( $R^{2}$=0.9978). The newly hatched pre-larvae was 4.9 mm in TL with ellipsoid yolk. In 16 days after hatching, larvae attained 6.8 mm in TL, and absorbed the yolk completely to become post-larval stage, but first heterotrophic food could be in 7 days after hatching already. Post-larval stage continued during 16~52 days after hatching with development of organs attachment. In 61 days after hatching with 41.3 mm in TL, the fries became a juvenile stage respectively having small teed lateral line, and a black blotch on the flank same as adults, but chin barbel was not developed yet. It was presumed that haddock changed food and ecological behavior after metamorphosis ken this time.e.

• Fisheries and Aquatic Sciences
• /
• v.24 no.11
• /
• pp.375-382
• /
• 2021

In this study, the reproductive behavior and embryonic and larval development of the short ninespine stickleback Pungitius kaibarae was described and illustrated based on observations during spawning, hatching, and larval rearing trials. Adult P. kaibarae were collected downstream in Jinhae during the reproductive season (April-May). Males had nuptial coloration on their entire black bodies, with blue dorsal spines and yellow eyes, whereas females had a brown spotted pattern on their bodies. Males built nests on the stems of water weeds and attracted females. Fertilization occurred in the nest immediately after spawning, and males guarded the eggs until hatching. The fertilized eggs of P. kaibarae were spherical, demersal, adhesive, and transparent, and each egg measured 1.43 ± 0.07 mm in diameter. The morula, blastula, and gastrula stages, as well as hatching began at 5, 18.5, 21.5, and 96 post fertilization (HPF), respectively, at 20.0 ± 0.5℃. The newly hatched larvae had a total length (TL) of 5.67 ± 0.50 mm, with a yolk volume of 0.583 ± 0.059 mm³. Their mouths and anuses had not yet opened. At 2 days posthatching (days post hatching, DPH), the yolk was completely absorbed and the larvae began to feed exogenously. Pigmentation was observed in freshly hatched larvae 4 h after hatching, with the presence of eight areas with a dotted pattern on the dorsal surface of the larvae and dispersed spots on the head and yolk sac. At 30 DPH, the TL of the juveniles was 21.34 ± 1.70 mm. The nest area and number of eggs were 259.56 ± 101.39 mm² (75.18-506.04) and 155.33 ± 114.12 (0-437), respectively.

• Development and Reproduction
• /
• v.18 no.4
• /
• pp.241-249
• /
• 2014

Hyphessobrycon eques is a famous fish for ornamental fish market and aquarium. They are inhabit in regions of Amazon and Paraguay River basin. Serpae fishs were investigated 2-3 males are chased to female, and then males attempted to simulate the females abdomen. After fertilization, eggs were kept in incubators at $28^{\circ}C$. The fertilized eggs had adhesive and demesal characteristics and had a mean diameter of $0.92{\pm}0.01mm$. Larvae hatched at 16 hrs post fertilization. The hatched larvae averaged $2.90{\pm}0.16mm$ in total length ($L_T$). Complete yolk sac resorption and mouth opening occurred on the third day post hatching. At 45 days post hatching, the larvae were $12.5{\pm}1.60mm$ $L_T$ and had reached the juvenile stage.