• Korean Journal of Animal Reproduction
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• v.20 no.4
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• pp.395-412
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• 1997

Mammalian eggs are ovulated arrested at meiotic metaphase II until fertilization. Generally in mammals, fertilization results in a series of intracellular calcium oscillations that are mediated by inositol triphosphate (IP$^3$) or cyclic adenosine diphosphoribose (cADPr). The high levels of maturation promotion factor (MPF) within the cell decrease, pronuclei form, the cytoskeleton is reorganized and proteins are post-translationally modified. If all is normal, the newly formed embryo initiates the developmental program specific to that species. Artificial methods of producing these effects in pig oocytes are discussed. One potential mechanism mediated via a signal transduction pathway is present in pig oocytes. Stimulation of this pathway leads to the early events following fertilization, and electrical stimulation leads to apparently normal de v velopment to day 12. Further studies are needed to determine which mechanism(s) the sperm uses to initiate development.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.111-115
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• 2008

The aim of this study was to examine more effective cryoprotectant for the cryopreservation of mouse preantral follicles. Enzymetically isolated preantral follicles from 12-day-old mice were cryopreserved by a slow freezing protocol with 1.5 M propanediol (PROH), dimethyl sulphoxide (DMSO) or glycerol (GLY) and then grown and matured in vitro for 11 days after thawing. The survival of preantral follicles immediately after freezing and thawing was not different among the PROH (68.2%), DMSO (72.4%) and GLY (72.1%). After grown and matured in vitro, the rates of survival and metaphase II oocytes were 54.9% and 36.6% for PROH which was significantly higher rates (p<0.05) compared with the rates obtained from DMSO (16.9% and 9.0%) and GLY (16.3% and 7.5%). The diameter of metaphase II oocytes from pre antral follicles frozen in PROH ($67.4{\pm}1.8\;{\mu}m$) was significantly (p<0.05) smaller than that of the fresh preantral follicles ($69.1{\pm}2.3\;{\mu}m$). The results from the present study revealed that PROH is more suitable cryoprotectant for the cryopreservation of mouse preantral follicles.

• Journal of Veterinary Clinics
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• v.7 no.2
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• pp.517-519
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• 1990

In vitro maturation and fertilization of oocytes collected from slaughtered bovine ovaries were investigated. Immature bovine extrafollicular oocytes were cultured for 24 hrs. in TCM 199 supplemented with fetal calf serum in a humidified CO$_2$ incubator. Fertilization in vitro was performed using frozen-awed bull semen which was treated by Ca Ionophore A23187. Fourty percentage of oocytes cultured had matured to the metaphase II ; There were-no effects of the concentration of fetal calf serum and of the addition of HEPES on the maturation rate. The mean proportions of in vitro fertilized eggs and of cleaved eggs were 23.1％ and 14.4％, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.1
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• pp.15-21
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• 2017

Objective: The aims of this study were to investigate whether fertilization could induce the resumption of meiosis in mouse oocytes arrested at metaphase I (MI) after in vitro maturation (IVM), and to investigate the effect of $Ca^{2+}$ chelator treatment at the time of fertilization on the transition from MI to metaphase II (MII). Methods: MII-stage and arrested MI-stage mouse oocytes after IVM were fertilized, and then embryonic development was monitored. Blastocysts from each group were transferred into 2.5 days post-coitum pseudo-pregnant ICR mice. MI oocytes after IVM were treated with a $Ca^{2+}$ chelator to investigate the effect of $Ca^{2+}$ oscillations on their maturation. Results: As insemination time increased, the number of oocytes in the MI group that reached the MII stage also increased. The blastocyst rates and total cell numbers in the MII group were significantly higher than in the MI group. No pregnancy occurred in the MI group, but 10 pregnancies were achieved (10 of 12) in the MII group. The proportion of MI oocytes that matured to MII oocytes after fertilization was significantly higher in the non-treated group than in the $Ca^{2+}$ chelator-treated group. Conclusion: The findings that a higher proportion of MI-arrested oocytes progressed to MII after fertilization and that the MI-to-MII transition was blocked by $Ca^{2+}$ chelator treatments before fertilization indicate that the maturation of MI oocytes to MII oocytes is associated with intracellular $Ca^{2+}$ oscillations driven by fertilization.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.357-366
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• 1996

The present experiments aimed to investigate the metabolism of calcium during oocyte maturation in rat. The concentration of free calcium and calmodulin in oocytes was measured respectively by using of fluo-3/AM and FITC with microscope fluorescence spectrometer. The ultrastructural localization of calcium precipitates in oocytes was observed with the transmission electron microscope. Cumulus-free immature oocytes(GV-oocyte) were cultured in vitro through 15 hours. The free calcium concentration in GV oocyte was $55.9{\pm}3.5nM$. In calcium-containing medium, the free calcium concentration was increased in germinal vesicle breakdown(GVBD) oocyte($64.2{\pm}7.3nM$). In normal medium after calcium chelator treatment ($10{\mu}M$ BAPTA/AM), the free calcium contents were slightly lower than those in control group. In calcium-free medium, the free calcium content was drastically increased in GVBD($72.7{\pm}3.4nM$) and metaphase I - anaphase I ($88.0{\pm}3.4nM$) oocyte. In maturation rate of oocytes, GVBD rate was high in control group($82.9{\pm}6.55%$) and calcium chelator treatment group($91.2{\pm}4.4%$), but in calcium-free medium group, it was low and then the oocyte was degenerated without polar body formation. Relative content of calmodulin in oocyte was significantly(P<0.001) increased in metaphase I - anaphase I than in GV and GVBD oocyte. The calcium precipitates were observed in mitochondria and cytoplasm of GV oocyte but that were not observed in mitochondria of GVBD and metaphase I - anaphase I oocyte. And then the calcium precipitates reappeared in mitochondria of metaphase II oocyte. The above results indicate that changes in free calcium and calmodulin concentration of oocyte occur according to the maturational stages and the extracellular calcium is required during oocyte maturation. Also change of calcium localization in oocyte occurs according to the maturational stages.

• Journal of Korean Society of Forest Science
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• v.45 no.1
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• pp.51-61
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• 1979

The chromosome behavior and it's synapsis in the meiosis of pollen mother cell were studied on Populus alba L. as a female parent tree, Populus glandulosa Uyeki as a male parent tree and their hybrid, Populus alba x glandulosa. 1. At metaphase I, the number of nuclear plates with early separation chromosome were observed with the lowest proportion of 11.0% in Populus glandulosa and with the highest proportion of 13.0% in Populus alba${\times}$glandulosa. 2. At metaphase II, early separation chromosomes appeared with the frequency of 11.0% in Populus alba${\times}$glandulosa. But the frequency was not different with those of the parental trees. 3. At anaphase I, lagging chromosomes appeared with some high rate of 11.6% in Populus alba${\times}$glandulosa and yet the number of chromosome bridges in populus alba x glandulosa almost were not different with the partental trees. 4. At anaphase II, lagging chromosomes appeared with some high frequency of 10.2% in Populus alba${\times}$glandulosa and the chromosome bridges in Populus glandulosa appeared with the highest frequency in all studied trees. 5. The frequency of abnormal pollen sporad was the highest value of 8.2% in Populus alba${\times}$glandulosa among the studied trees. With the results, it might be assured that the chromosome segregation and it's synapsis behaved normally in Populus alba, Populus glandulosa and Populus alba x glandulosa, and so all the studied trees could produced normal pollens.

• Journal of Korean Society of Forest Science
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• v.51 no.1
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• pp.1-21
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• 1981

Populus glandulosa and Populus tomentiglandulosa, which were known to be natural hybrids, were examined for morphological, physiological and karyological traits to illucidate its hybridity and taxonomical importance. The results abtained were as follows; 1. Survival rate in rooting of cuttings and grafting was different between the hybrids and their rooting abilities showed incomplete dominance. 2. Their leaf openings showed incomplete dominance. The leaf longevities of P. alba ${\times}$ glandulosa and P. tomentiglandulosa were stronger than the other hybrids. 3. There were differences in resistance to toxicity of $KClO_3$ between the hybrids. 4. Many external leaf characters of the hybrids also showed incomplete dominance. P. tomentiglandulosa was similar in those characters to P. alba ${\times}$ glandulosa while P. glandulosa was similar to hybrids crossed, reciprocally crossed or back-crossed between P. davidiana and P. alba. 5. Their numbers of male flower showed incomplete dominance or hybrid vigor. The numbers of P. tomentiglandulosa were similar to thosa of P. alba ${\times}$ glandulosa while those of P. glandulosa to those of P. alba ${\times}$ davidiana or P. davidiana ${\times}$ alba. 6. Morphology and band color of male catkin bract showed incomplete dominance. Those of P. glandulosa were similar to those of P. alba ${\times}$ davidiana while those of P. tomentiglandulosa to those of. P. alba ${\times}$ glandulosa. 7. There were differences in vascular bundle number and arrangement of petiole between the hybrids. 8. Differences in the anatomical traits of stem did not exist between the hybrids but those in wood fiber size existed. 9. The chromosomes of artificial hybrids, P. glandulosa and P. tomentiglandulosa showed irregular behavior in metaphase I and II. 10. All hybrids including P. glandulosa and P. tomentiglandulosa showed small number of P.M.C. with 19 II but many univalent chromosomes were exhibited in metaphase I. 11. All hybrids including P. glandulosa and P. tomentiglandulosa showed a little abnormal nuclear plates as laggard chromosome and chromosome bridge in anaphase I and II. 12. The frequency of pollen tetrad and fertile pollen was low in most of the hybrids including P. glandulosa and P. tomentiglandulosa.

• Development and Reproduction
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• v.8 no.1
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• pp.35-42
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• 2004

Oocyte maturation refers to the process that prophase I arrested germinal vesicle(GV) drives the progression of meiosis to metaphase II(MII) to have the capacity for fertilization and embryo development. To better understand the molecular mechanism(s) involved in oocyte maturation, we identified differentially expressed genes(DEGs) between GV and MII mouse oocytes using a new innovative annealing control primer (ACP) technology. Using 20 ACPs, we successfully cloned 32 DEGs between GV and Mll oocytes, and 26 out of these 32 DEGs were functionally known genes. Four genes including Pscd2 were GV-specific, 10 genes including PKD2 and CSN3 were highly expressed in GV oocytes(GV-selective), and 12 genes including Diva were highly expressed in MII oocytes (MII-selective). Ail of the genes identified in this study were first reported in the oocyte expression using ACP system and especially, we could characterize the existence of PKD-CSW signaling pathwayin the mouse oocytes. Results of the present study would provide insight for studying molecular mechanisms regulating oocyte maturation.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.3
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• pp.317-324
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• 2004

The present study was conducted to investigate the involvement of nitric oxide (NO) in cumulus expansion, oocyte mortality and meiotic maturation of porcine cumulus enclosed oocytes (CEOs) cultured in two different models when gonadotropins, including follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG) were presented or not. And the interaction between NO and $\beta$-mercaptoethanol ($\beta$-ME), a free radical scavenger was also investigated. Two models refer to spontaneous maturation model and hypoxanthine (HX) medium model. All the 3,433 eligible CEOs were incubated at $39^{\circ}C$ and the cumulus expansion, oocyte morphology and nuclear phase were evaluated 44 h after incubation. (1) In spontaneous maturation model, NO stimulates the cumulus expansion and $\beta$-ME delayed it. NO doesn't affect the oocyte meiotic resumption but inhibits the oocytes to develop to metaphase II. (2) In HX medium model, NO or $\beta$-ME doesn't affect the expansion in the absence of gonadotropins, but in the presence of gonadotropins, NO or $\beta$-ME inhibits the expansion. In the presence of gonadotropins, NO inhibits the oocyte meiotic resumption and it especially inhibits the oocyte to develop to metaphase II, and $\beta$-ME reverses such inhibitory effects. The cooperation of gonadotropins and $\beta$-ME stimulates the meiotic resumption and especially, promotes the CEOs to develop to metaphase II in both models. Moreover, HX might contribute to the fragility of oocyte zona pellucida and gonadotropins, nitric oxide and $\beta$-ME could alleviate it separately, and cooperatively. It is concluded that NO exerts different functions in two models and $\beta$-ME affected the functions of NO in different models.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.53-61
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• 1999

The present study was designed to investigate the effects of gonadotrophins on in vitro growth and maturation in mouse preantral follicles. Ovaries were removed from 12-day-old ICR mice. Follicles were dissociated enzymetically in Leibovitz L-15 medium containing 1 mg/$m\ell$ collagenase and 0.2 mg/$m\ell$ DNase I. The follicles were cultured on Transwell-COL membrane inserts in six well cluster dishes for 10 days. The culture medium was $\alpha$MEM medium supplemented with 5% fetal bovine serum and FSH or HMG. After 10 days of growth in vitro, follicles were allowed to mature for 18~20 hr in medium supplemented with 1.5 IU/$m\ell$ hCG. The oocytes were then denuded of their cumulus cells and assessed maturation status. Concentrations of oestradiol and progesterone were measured with a radioimmunoassay. Oocyte diameter was determined with an ocular micrometer. The survival and Metaphase II rates of oocytes were significantly higher in FSH treatment groups than in control group (P＜0.001), but there were no differences among the groups of treated FSH concentration. The survival and Metaphase II rates of oocytes in HMG treatment group (60.9 and 40.6%) were higher than in FSH treatment group (76.6 and 48.2%) and control group (49.2 and 7.1%). The survival and Metaphase II rates of oocytes on both FSH and LH treatment groups were no differences among the ratios of FSH and LH. Diameter of oocyte was no differences among the treatment groups, but smaller than compared to in vivo grown oocyte. Through the entire culture period, secretions of oestradiol and progesterone were significantly less in control group than in HMG and FSH treatment groups. These results suggest that gonadotrophins playa key role in in vitro culture of mouse preantral follicles. Especially, addition of FSH and LH should be more effective than FSH alone.