• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.2
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• pp.139-143
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• 2005

The implications of temperature (25, 30 and $35^{\circ}C$) and relative humidity (RH; 60, 70 and $80\%$) on the hatching rhythmicity and hatching parameters (percentage and duration) were studied in the silkworm, Bombyx mori L. under natural photoperiod (LD 12 : 12). Disease free layings (DFLs) of two pure silkworm breeds, Pure Mysore (PM, a multivoltine breed) and $NB_4D_2$ (a bivoltine breed), and their hybrid, $PM{\times}NB_4D_2$ were introduced into the experimental conditions on the $3^{rd}$ day of oviposition till completion of hatching. The hatching rhythm was predominantly diurnal under all temperature and humidity conditions, with peaks just after 'lights-on' phase (6 hrs). Extreme temperature and humidity conditions did not alter the hatching rhythmicity, but prolonged the hatching durations, extending it to the next day, coupled with reduced hatching percentage in PM and $PM{\times}NB_4D_2{\cdot}In\;NB_4D_2$, on the other hand, hatching did not extend to the next day. Hatching percentage in this breed, however, reduced below the economic level under high temperature and low humidity conditions. The high temperature and low humidity together, though did not alter the rhythmicity, seems to exert synergetic effect on the hatching percentage and its duration in the silkworm, B. mori.

• Development and Reproduction
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• v.16 no.1
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• pp.69-75
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• 2012

Hatching occurred in the time dependent manners and strictly controlled. Although, the hatching processes are under the control of muti-embryotrophic factors and the expressed G proteins of cell generate integrated activation, the knowledge which GPCRs are expressed during hatching stage embryos are very limited. In the present study, which G proteins are involved was examined during blastocyst development to the hatching stage. The early-, expanded-, and lobe-stage blastocysts were treated with various $G_{\alpha}$ activators and H series inhibitors, and examined developmental patterns. Pertusis toxin (PTX) improved the hatching rate of the early-stage blastocyst and lobe-formed embryos. Cholera toxin (CTX) suppressed the hatching of the early-stage blastocyst and expanded embryos. The effects of toxins on hatching and embryo development were changed by the H7 and H8. These results mean that PTX mediated GPCRs activation is signaling generator in the nick or pore formation in the ZP. In addition, PTX mediated GPCR activation induces the locomotion of trophectoderm for the escaping. CTX mediate GPCRs activation is the cause of suppression of hatching processes. Based on these data, it is suggested that various GPCRs are expressed in the periimplantation stage embryos and the integration of the multiple signals decoding of various signals in a spatial and temporal manner regulate the hatching process.

• Journal of Sericultural and Entomological Science
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• v.20 no.2
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• pp.6-9
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• 1978

This experiment was attempted to investigate the effect of the specific gravity of silkworm eggs on the hatching and the practical hatching ratio depend upon six egg production companies for two silkworm races and their reciprocal crosses. 1. The hatching and the practical hatching ratio in the specific gravity of medium and heavy eggs were higher than in the specific gravity of light eggs. 2. Compare with Japanese or Chinese mother races each other, it was inclined that the former seems to be higher than the latter on the hatching ratio in the specific gravity of light and medium eggs, but the practical hatching ratio was high only in the specific gravity of light eggs. 3. Chinese mother races were different in the practical and the hatching ratio between the specific gravity of eggs. On the contrary in case of Japanese mother races were no difference for the hatching ratio but difference in the practical hatching ratio between the specific gravity of eggs. 4. On the egg production company, in case of the specific gravity of medium and light eggs, the hatching and the practical hatching ratio were high significance, but no difference in the specific gravity of heavy eggs.

• Fisheries and Aquatic Sciences
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• v.14 no.4
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• pp.417-420
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• 2011

In this experiment, we examined the survival, fertilization, hatching times, and hatching rates of Far Eastern catfish Silurus asotus at pH ranging from 2 to 13 under laboratory conditions. Eggs could be fertilized at pH 3-12. In a hatching experiment, mortality was first observed at pH 13, when all fertilized eggs died within 8 min, followed by pH 2 (30 min), pH 12 (60 min), pH 3 (4 h), and pH 11 (5 h). Hatching only occurred at pH 4-10, with the highest hatching rate at pH 7 (52%) and the lowest at pH 10 (24%). Hatching rates in acid solutions were higher than in alkaline solutions, although the difference was not significant. Hatching was first observed at pH 10, beginning 27 h after fertilization and ending at the 31 h. A clear difference was observed between hatching times, ranging from 31 to 64 h and increasing in order with descending pH.

• Animal cells and systems
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• v.7 no.4
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• pp.345-348
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• 2003

The effects of acute acid stress on hatching success and hatching period of laboratory-reared hermaphroditic fish Rivulus marmoratus were examined. The effects of acute acid toxicity on mortality was also determined in three life stages of this fish. There was a significant negative effect of acid stress on hatching performance in the R. marmoratus embryos. The hatching success was only 5% at pH 3.5 compared to over 78% at pH higher than 4.0. The hatching period was also delayed by low pH treatments. The larval and juvenile stages were more sensitive to acid toxicity on mortality than the adult stage, but larvae and juveniles showed similar sensitivity. The 96-h LC50 value was pH 3.8 in larval and juvenile stages and pH 3.3 in adult stage.

• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.75-79
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• 2006

The developmental regulation of the preimplantation mammalian embryos is a fundamental step for preparing the implantation and it may be regulated by several autocrine and paracrine factors including platelet-activating factor. PAF improved the embryonic survival and implantation but its role during blastocyst development is still largely unknown. In this study, the effects and the possible pathway of PAF on developmental regulation of blastocyst to hatching stage were investigated. Developmental pattern in hatching embryo was a concentration-response curve showing maximal activity at 1 nM PAF, with decreasing activity at higher concentrations. $50{\mu}M$ 1-(5-isoquinolimnesulfonyl)-2-methylpiperazinme dihydrochloride (H-7), a PKC inhibitor, inhibited the progression of blastocyst to hatching embryo. In addition H-7 blocked the PAF effects on the blastocyst development. Besides tetradecanoylphorbol acetate (TPA), a PKC activator stimulated development of blastocyst to the hatching stage. These finding revealed that PAF support the blastocyst development to the hatching embryo. Also it is suggested that PAF action pathways in hatching supporting include the PKC signaling pathway.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.147-153
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• 2001

Objective: The present study was performed to investigate the efficiency of partial laser assisted hatching (p-LAH; lased 1/2 ZP width from ZP edge) on hatching of mouse blastocysts. Methods: We used non-contact $1.48{\mu}m$ diode laser (MTM, Switzland) to create a precise hole on zona pellucida. 2-cell embryos were collected from the mouse (ICR) oviduct at 48 hours after hCG administration. Collected 2-cell embryos were cultured in the P-1 medium supplemented with 0.4% BSA. For experiments, embryos at 8-cell stage were used after $20{\sim}22$ hours in culture. After conventional (c-LAH) or partial laser assisted hatching, the embryos were further cultured in P-1 medium supplemented with 0.4% BSA for 3 days. To compare efficiency of complete and partial laser assisted hatching, hatching rate, hatching time and blastocyst diameter and zona pellucida thickness at hatching time were investigated. Embryos were examined every 12 hours. Blastocyst diameter and zona pellucida thickness at hatching time were measured with an ocular micrometer. Results: Hatching rates of p-LAH group (84.2%) was significantly higher than that of control group (39.3%), but there was no difference between the p-LAH (84.2%) and c-LAH (91.2%). p-LAH group was hatched 12 hours earlier than control group, but hatched 12 hours later than c-LAH group. The diameter of blastocyst at hatching time of p-LAH group ($113.1{\pm}6.4{\mu}m$) was smaller than that of control group ($122.2{\pm}5.0{\mu}m$), but larger than that of c-LAH group ($102.2{\pm}2.7{\mu}m$). Zona pellucida thickness at hatching time of p-LAH group ($6.4{\pm}0.9{\mu}m$) was thicker than that of control group ($4.5{\pm}1.5{\mu}m$), but thinner than that of c-LAH group ($10.0{\pm}0.8{\mu}m$). Conclusion: These results suggest that p-LAH may maintains the cell arrangement of early embryos to ensure successful development and prevent precocious hatching of blastocyst when compare to c-LAH and conventional (acidic tyrode) AH. Thus, p-LAH may provide a valuable and effective AH technique for human ART program.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.35-42
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• 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
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• v.41 no.2
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• pp.68-74
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• 2014

Objective: In search of an ideal method of assisted hatching (AH), we compared the effects of conventional micropipette-AH and laser-AH on the blastocyst formation rate (BFR) and blastocyst cell numbers. Methods: Four-to five-week-old ICR female mice were paired with male mice after superovulation using Pregnant mare's serum gonadotropin (PMSG) and hCG. The two-cell embryos were flushed from the oviducts of female mice. The retrieved two-cell embryos underwent one of five AH procedures: single mechanical assisted hatching (sMAH); cross mechanical assisted hatching (cMAH); single laser assisted hatching (sLAH); quarter laser assisted hatching (qLAH); and quarter laser zona thinning assisted hatching (qLZT-AH). After 72 hours incubation, double immunofluorescence staining was performed. Results: Following a 72 hours incubation, a higher hatching BFR was observed in the control, sMAH, cMAH, and sLAH groups, compared to those in the qLAH and qLZT-AH groups (p<0.05). The hatched BFR was significantly higher in the qLAH and qLZT-AH groups than in the others (p<0.05 for each group). The inner cell mass (ICM) was higher in the control and sMAH group (p<0.05). The trophectoderm cell number was higher in the cMAH and qLAH groups (p<0.05). Conclusion: Our results showed that the hatched BFR was higher in groups exposed the the qLAH and qLZT-AH methods compared to groups exposed to other AH methods. In the qLAH group, although the total cell number was significantly higher than in controls, the ICM ratio was significantly lower in than controls.

• Journal of Sericultural and Entomological Science
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• v.10
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• pp.59-62
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• 1969

It is very important physiological and economical problems to hatching within a day by increasing hatching-ratio of silkworm eggs in sericulture, and modified dark incubating method was experimented in Japan. The author studied on the economical problems of that rather than physiological study. Hatching was induced and accelerated by illumination before hatching 5 days which incubated in the dark room after eye spot pigmented eggs emerged 10-20 percentage. It was increased by 80-90％ in hatching-ratio, but the hatching was delayed a day. In autumn silkworm, newly hatched silkworms were hatched silkworms were hatched in the dark room because of high temperature period tough modified dark incubation was passed only 4 days. The author wonder which is more effective between effect of increasing the hatching-ratio and the health of silkworms.