• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.343-351
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• 1996

The objectives of this study are to determine cortical granule distribution during in vitro maturation, parthenogenetic activation and in vitro fertilization of oocytes, and to investigate effects of microfilament inhibitor on the cortical granule distribution during in vitro maturation and fertilization of oocytes in the pig, The corti-cal granule distribution were imaged with fluor-escent labeled lectin under laser scanning confocal microscope or detected by transmission electron microscope. At germinal vesicle stage, cortical granule organelles were located around the cell cortex and were present as a relatively thick area on the oolema. Microfilaments were also observed in a thick uniform area around the cell cortex. Following germinal vesicle break down,microfilaments concentrated to the condensed chromatin and cortical granules were observed in the cortex. Treatment with cytochalasin B inhibited microfilament polymerization and prevented movement of cortical granules to the cortex. Cortical granule exudate following sperm penetration was evenly distributed in the entire perivitelline space. Therefore, these results suggested that the microfilament assembly is involved in the distribution, movement and exocytosis of cortical granules during maturation and fertilization of porcine oocytes. (Key words cortical granule, porcine, maturation, fertilization).

• Development and Reproduction
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• v.2 no.2
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• pp.157-163
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• 1998

Preimplantation embryos of the rat was examined by the morphological changes in the cortical granule envelope (CGE), blastomere surface, and zona pellucida (ZP) of embryo after cortical reaction. The ultrastructural characteristics and CGE of embryos were observed with the scanning electron microscope and fluorescence microscope. In the ultrastructural characteristic of embryo surface, surface microvilli were shortened and the CGE-like structure existed above microvilli in eight-cell embryo. Rough spongy surface and decreased network numbers were key characters of embryonic ZP compared to unfertilized oocyte. The CGE formed by cortical reaction existed in perivitelline space during embryo development but it was thin and locally distributed ill contrast to fertilized oocyte. The present results indicate that cortical reaction forms cortical granule envelope in perivitelline space and causes not only zona hardening, but also ultrastructural changes in ZP and cell membrane of preimplantation embryos.

• The Journal of Korean Medicine
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• v.23 no.1
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• pp.120-132
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• 2002

Objectives : The purpose of this investigation was to evaluate the effect of Goomicheongsim-won Extracts on E20 corticells and P7 cerebellar cells exposed to hypoxia, and the effect on neuronal protection by elimination of Rhinoceros unicornis L. and/or Orpiment $As_2S_3$. Methods : P7 cerebellar cells were grown in various concentrations of KM-A, KM-B, KM- C and KM-D. On 7 DIV (day in vitro), cells were exposed to hypoxia (98% $N_2/5%{;}CO_2,{\;}3{\;}hr,{\;}37^{\circ}C$) and normoxia, and then further incubated for 3 days. Neuronal viabilities were expressed as percentages of control. E20 cortical cells were grown in various concentrations of KM-A, KM-B, KM-C, and KM-D. On 7 DIV, cells were exposed to hypoxia and normoxia, and then further incubated for 3 and 7 days. Results : I. The effect of KM-A on neuronal protection was significantly increased P7 cerebellar granule cells and E20 cortical cells on normoxia and hypoxia. 2. The effect of KM-B on neuronal protection was increased P7 cerebellar granule cells on normoxia, but was significantly decreased P7 cerebellar granule cells on hypoxia. The effect of KM-B on neuronal protection was non-significantly increased E20 cortical cells on normoxia and hypoxia. 3. The effect of KM-C on neuronal protection was non-significantly increased P7 cerebellar granule cells on normoxia and hypoxia and was decreased (p=0.058) on hyperconcentration of the extracts in normoxia. The effect of KM-C on neuronal protection was significantly increased P7 cerebellar granule cells and E20 cortical cells on normoxia and hypoxia (10 DIV), and the effect was E20 cortical cells on normoxia (14 DIV), non-significantly increased E20 cortical cells on hypoxia (14DIV). 4. The effect of KM-D on neuronal protection was increased P7 cerebellar granule cells on normoxia but was not on hyperconcentration of the extracts, was significantly decreased on hyperconcentration of the extracts in hypoxia. The effect of KM-D on neuronal protection was significantly increased E20 cortical cells on normoxia and was significantly increased E20 cortical cells increased on hypoxia (10 DIV). Conclusions : Goomicheongsim-won extracts had applicable effect on E20 corticells and P7 cerebellar cells exposed to hypoxia. The effect on neuronal protection by elimination of Rhinoceros unicornis L. and/or Orpiment $As_2S_3$ was changed.

• Development and Reproduction
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• v.3 no.1
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• pp.21-28
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• 1999

In order to examine the effect of $Ca^{2+}$ -chelation on in vitro fertilization of rat zona-free oocyte, the formation of cortical granule envelope (CGE) and the rate of fertilization related to monospermy and polyspermy were determined. The ultrastructural characteristics of oocytes were observed with the scanning electron microscope and BAPTA/AM was used for calcium-chelation. The CGE formed by cortical reaction was observed in zona-free oocyte inseminated in vitro and it was also observed in the calcium chelator (1, 5, 10$\mu$M BAPTA/AM) treated zona-free oocytes inseminated in vitro. The CGE developed according to incubation time. The fertilization rate was decreased in the calcium chelator-treated group (59.8, 38.1, 37.0%) compared to the control group (60.6%) but monospermy rate was increased in the calcium chelator-treated group (45.0, 47.3, 50.9%) compared to control group (37.5%). The above results demonstrate that the CGE is formed during fertilization in rat and the extracellular calcium is used in cortical reaction. Also the results suggest that proper concentration of free calcium in oocyte acts as important factor in fertilization.n.

• Journal of Embryo Transfer
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• v.17 no.3
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• pp.239-249
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• 2002

Embryos derived from pig oocytes matured in mSOF are able to develop to blastocysts after IVF. Experiment 1 evaluated the effects of two maturation media (TCM-199 vs mSOF) on maturation rate, fertilization parameters, including penetration, polyspermy, male pronuclear formation, and the mean number of sperm penetrated per oocyte. Experiment 2 and Experiments 3 examined the effects of two maturation media on zona pellucida solubility and cortical granule distribution by transmissible electron microscopy, respectively. Experiment 4 assessed the effects of two maturation media on the in vitro embryo cleavage rate and development to blastocyst. Lastly, experiment 5 examined the cell number of blastocyst. An effect of media (P<0.05) was detected for mSOF on the mean number of sperm per oocyte. In TCM group, zona digestion time (196.5$\pm$15.5 vs 131.6$\pm$20.1 before IVF, 397.5$\pm$30.3s vs 185.3$\pm$16.4s after IVF, p<0.05) was higher in TCM-199 group. No significant effects of media was observed on cortical granule distribution between two groups by TEM. An effect (P<0.05) was observed on embryo development to blastocyst (16% vs 8%) but not on cleavage rates. No significant effects of media was observed on total cell number of blastocyst. We found that the high mean number of sperm penetrated per oocyte and the weaker zona pellucida on the basis of the digestion time was shown in pig oocytes matured in mSOF, however, porcine oocyte maturation with supplemented synthetic oviduct fluid medium (mSOF) resulted in blastocyst cell numbers comparable to those observed with Tissue Culture Medium 199.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.07a
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• pp.52-52
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• 2005

Oogenesis, the gonadosomatic index (GSI), reproductive cycle and first sexual maturation of the female Neptunea (Barbitonia) arthritica cumingii have been investigated by light and electron microscope observations. In the early vitellogenic oocyte, the Golgi complex and mitochondria were involved in the formation of glycogen, lipid droplets and yolk granules. In late vitellogenic oocytes, the rough endoplasmic reticulum and multivesicular bodies were involved in the formation of proteid yolk granules in the cytoplasm. In particular, compared with the results of other gastropods, it is a different result that appearances of cortical granules at the cortical layer and microvilli on the vitelline envelope, which is associated with heterosynthetic vitellogenesis, were not observed in vitellogenic oocytes during oogenesis. A mature yolk granule was composed of three components: main body (central core), superficial layer, and the limiting menbrane, Monthly changes in the gonadosomatic index in females were studied in 2002 and 2003 were closely associated with ovarian developmental phases. Spawning occurred between May and August in 2002 and 2003 and the main spawning occurred between June and July when the seawater temperature rose to approximately 18${\sim}$23${\circ}$C. The female reproductive cycle can be classified into five successive stages: early activestage (Septmber to October), late active stage ( November to February), ripe stage (February to June), partially spawned stage (May to Aygust), and recovery stage (June to August).

• The Korean Journal of Malacology
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• v.22 no.1 s.35
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• pp.1-11
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• 2006

The gonadosomatic index (GSI), oogenesis and reproductive cycle in female Neptunea (Barbitonia) afhritica cumingii were investigated by light and electron microscope observations. In the early vitellogenic oocyte, the Golgi complex and mitochondria were involved in the formation of glycogen, lipid droplets and yolk granules. Late in the vitellogenic oocytes, the rough endoplasmic reticulum and multivesicular bodies were involved in the formation of proteid yolk granules in cytoplasm. In particular, compared with the results of other gastropods, it showed a different result that appearances of cortical granules at the cortical layer and microvilli on the vitelline envelope, which is associated with heterosynthetic vitellogenesis, were not observed in vitellogenic oocytes during oogenesis. A mature yolk granule was composed of three components: main body (central core), superficial layer, and the limiting membrane. Monthly changes in the gonadosomatic index in females studied in 2004 and 2005 were closely associated with ovarian developmental phases. Spawning occurred between May and August in 2004 and 2005 and the main spawning occurred between June and July when the seawater temperature rose to approximately $18-23^{\circ}C$. The female reproductive cycle can be classified into five successive stages: early active stage (September to October), late active stage (November to February), ripe stage (February to June), partially spawned stage (May to August), and recovery stage (June to August).

• Animal Systematics, Evolution and Diversity
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• v.21 no.1
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• pp.45-56
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• 2005

Three species of Gastrostyla ciliates collected from the sewage treatment plant and a puddle in the valley from Korea were identified as Gastrostyla minima Hemberger, 1985, G. steinii Engelmann, 1862 and G. setifera (Engelmann, 1862). The description was based on the observation of living specimens, protargol impregnated specimens and biometric analysis. The morphological variations among the populations of these species were investigated with morphometry. These species have not been reported in Korea and their diagnostic characteristics are as follows: G. minima normally has two oval macronuclei (Ma) with one spherical micronucleus (Mi) respectively, continuous ventral cirral row (VCR) with additional one postoral ventral cirrus (poVC), five transverse cirri (TC), six dorsal kineties (DK) with broken 4th kinety, and cortical granules. G. steinii has four oval Ma with three to five Mi, discontinuous VCR with additional one poVC, four TCs, six DKs, and no cortical granules. G. setifera has two oval Ma with one spherical Mi respectively, five TCs which distinctly separate in two groups, six DKs, discontinuous VCR with additional two poVCs, and no cortical granules.

• 대한생식의학회:학술대회논문집
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• 2007.06a
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• pp.127-127
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• 2007

• Applied Microscopy
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• v.30 no.1
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• pp.61-72
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• 2000

Three fertilization envelopes (FE) have been observed after the artificial insemination of U. unicinctus oocytes. The substances of the first fertilization envelope, which is an effective barrier against excessive sperm, come mainly from the surface coat of the oocyte. The secretions of the cortical granules take part in formation of the 2nd fertilization envelope. Histologically, the 3rd fertilization envelope is not amorphous as seen under light microscope, but contains numerous panicles under electron microscope, which would be contributed to harden the envelope by 60 min after the fertilization. With the substantial similarity between the 1st fertilization envelope and the surface coat of the oocyte, and the coincidence of retraction of microvilli and the formation of the 1st fertilization envelope, it is suggested that the microvilli contain the sperm receptors in U. unicinctus. Some granular substances from the distal part of the acrosome diffuse on the surface coat of the oocyte while the acrosomal tubules penetrate into the surface coat. The acrosomal tubules arise from the proximal part of the acrosome and pass through the acrosomal lumen.