• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.181-189
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• 2001

The objective of this study was to evaluate the development of porcine follicular oocytes fertilized by intracytoplasmic sperm injection (ICSI). Cumulus-oocyte-complexes (COCs) were collected by aspiration from follicles of 2~7 mm in diameter from a local slaughterhouse. Oocytes were matured in vitro for 40~44 h, and spermatozoa were prepared by swim-up in the presence or absence of 5 mM dithiothreitol (DTT) and then M II stages of the oocyte were either centrifuged or not centrifuged for the following injection of ooplasm. Injected oocytes were cultured in NCSU 23 medium for 6 to 8 days. The results obtained were as follows: 1. The rates of cleavage and development rates into blastocyst by ICSI were not significantly different between the with (53.0% and 19.7%) or without (48.3% and 23.8%) centrifugation, respectively (P＜0.05). 2. The cleavage and developmental rates to blastocyst after ICSI with or without 5 mM DTT treated-sperm were not significantly different (60.4% vs 16.4% and 45.5% vs 22.2%), respectively (P＜0.05). 3. The cleavage and the developmental rates to btastocyst were not significantly different between the zygotes obtained by IVF (51.8% vs. 22.4%) and ICSI (51.4% vs. 21.6%) (P＜0.05). 4. The number of blastomere in blastocyst stages after IVF or ICSI was not significantly different (46.7$\pm$2.9 and 41.9$\pm$4.6).

• Applied Microscopy
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• v.41 no.3
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• pp.189-196
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• 2011

Histological study on egg envelope and oogenesis of Kichulchoia multifasciata (Pisces, Cobitidae) was carried out by light microscopes and scanning electron microscopes. Various developmental cells appeared in ovary of the specimen catched during November 2010. The cytoplasm of oogonia was acidic and many nucleoli were located at the inner side of nucleus membrane. The size of the oogonia was $103.9{\pm}24.7$ ${\mu}m$ with nucleus size $42.9{\pm}6.9$ (31.1~50.3) ${\mu}m$. Primary oocyte having $277.5{\pm}60.5$ (216.7~354.9) ${\mu}m$ in diameter began to accumulate yolk vesicles. As the developmental stages proceed, secondary oocyte grows larger to $617.6{\pm}85.1$ (503.4~723.6) ${\mu}m$, and eosinophilic yolk granules yolk granules appeared between the yolk vesicles occupying most cytoplasm, and there are some yolk mass formed already. There are some yolk mass formed already. Envelope of fertilized egg investigated by a scanning electron microscope had plenty of microvilli (2~3 ${\mu}m$ in length) over the entire egg surface and a micropyle. Especially, the microvilli surrounding the micropyle were longer than those of egg surface with $5.26{\pm}1.22$ ${\mu}m$.

• The Korean Journal of Malacology
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• v.20 no.1
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• pp.55-63
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• 2004

Gonadosomatic index, condition index and reproductive cycle with the gonadal development of the female Octopus ocellatus were investigated by histological observations and morphometric data, from January to December, 2000. And changes in biochemical components of the ovary and the trunk tissues including the digestive organ associated with gonadal development were studied by biochemical analysis from January to October, 2001. The specimens were collected at the coastal waters of Buan, Jeollabuk-do, Korea, from January 2000 to October 2001. O. ocellatus is a dioecious organism. The gonad of O. ocellatus locates medially in posterior region of the body. Morphology of the ovary shows round and oval in shape, the average diameter and external colour of ripe ovary was 32 mm and semitransparent light brown in colour. As the ovary was getting mature, transparent elongated eggs covered with chorion were present in the ovarian cavity. Monthly changes in the gonadosomatic index (GSI) showed a similar pattern with those of the condition index. The GSI and condition index began to increase in March and reached the maximum in April. And then, their values decreased from May and reached the minimum in September. Reproductive cycle of O. ocellatus can be categorized into five successive stages: early developing stage (September to December), late developing stage (November to March), ripe stage (March to May), partially spawned stage (April to June), and degenerative/resting stage (June to October). Follicle cells attached to an oocyte were involved in vitellogenesis in the cytoplasm of the vitellogeneic oocyte and formation of chorion (secondary egg membrane) of the ovarian eggs. Spawning occurred between April and June. The spawning period was once a year and the peak took place between May and June. This species belongs to semelparity. According to changes in biochemical contents of the ovary and the digestive organ, monthly variations of moisture, total protein, total lipid and glycogen contents (%) in the ovary showed a negative correlationship with those of the trunk tissues including the digestive organ. Accordingly, it is assumed that the ovary only may be received nutrient supply (total lipid content) for gonadal development from the trunk tissues including the digestive organ (r = -0.55, p < 0.05).

• The Korean Journal of Malacology
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• v.27 no.1
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• pp.43-53
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• 2011

The gametogenic cycle and the spawning season in female and male Cyclina sinensis were investigated by quantitative statistical analysis using an image analyzer system, and the biological minimum size (the size at 50% of sexual maturity) was calculated by combination of quantitative data by size and von Bertalanffy's equation. Compared the gametogenic cycle by quantitative statistical analysis with the previous qualitative results in female and male C. sinensis, monthly changes in female and male gametogenic cycles calculated by quantitative statistical analysis showed similar patterns to the gonadal stages in female and male reproductive cycles by qualitative histological analysis. Comparisons of monthly changes in the portions (%) of each area to eight kinds of areas by quantitative statistical analysis in the gonads in female and male C. sinensis are as follows. Monthly changes in the portions (%) of the ovary areas to total tissue areas in females and also monthly changes in the portions of the testis areas to total tissue areas in males increased in March and reached the maximum in May, and then showed a rapid decrease from June to October. Monthly changes in the portions (%) of oocyte areas to ovarian tissue areas in females and also monthly changes in the portions of the areas of the spermatogenic stages to testis areas in males began to increase in March and reached the maximum in June in females and males, and then rapidly dropped from July to October in females and males when spawnig occurred. From these data, it is apparent that the number of spawning seasons in female and male C. sinensis occurred once per year, from July to October. Monthly changes in the number of the oocytes per mm2 and in the mean diameter of the oocyte in captured image which were calculated for each female slide showed a maximum in May and reached the minimum from December to February. Therefore, C. sinensis in both sexes showed a unimodal gametogenic cycle during the year. The percentage of sexual maturity of female and male clams ranging from 25.1 to 30.0 mm in length was over 50% and 100% for clams over 40.1 mm length. In this study, the biological minimum size (sexually mature shell lengths at 50% of sexual maturity) in females and males were 26.85 and 26.28 mm, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.2
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• pp.198-203
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• 1999

Gonadal development and reproductive cycle off Gomphina melanaegis collected in the coastal waters of Chumunjin, Korea were investigated monthly from April 1996 to April 1997. G. melanaegis was dioecious, The gonads were located between the digestive diverticula and muscle tissues of the foot, The ovary was composed of a number of ovarian sacs, and the testis was composed of several testicular tubules. The flesh weight rate was reached the maximum in August ($23.0\%$), and then decreased to $19.8\%$ in September. In March, the value was reached the minimum ($17.8\%$) and then increased, The size of mature oocyte was ranged $50\~60\mu$m in diameter and had a germinal vesicle with a nucleolus. Mature oocyte contained a large number of yolk granules and lipid granules in its cytoplasm. The spermatozoon was consisted of a conical nucleus with acrosome, a middle piece containing four mitochondria and proximal and distal centrioles, and a flagellum, Sex ratio (male/female) and minimum size for sexual maturation of G. melanaegis were 0.79 and about 25 mm in shell length, respectively. The reproductive cycle could be classified into five succesive stages: multiplicative (December to March), growing (April and May), mature(June), sprawning (July and August), and degenerative and resting (September to November) stages.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.8
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• pp.1190-1195
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• 2007

This study was conducted to improve efficiency of a follicle culture system without reducing developmental competence of intrafollicular oocytes. Preantral follicles (100 to $125{\mu}m$ in diameter) of F1 hybrid (B6CBAF1) mice were cultured singly for 216 h in modified ${\alpha}$-MEM-glutamax medium, to which 2.5 IU/ml hCG and epidermal growth factor was added 16 h prior to the end of culture. Medium change was either performed three times (54 h interval), twice (72 h interval), once (108 h interval), or not at all (216 h interval). Maturation (progression to the metaphase II stage) of intrafollicular oocytes was detected from 4 days after culture in the three-times change treatment, while all treatments yielded mature oocytes from day 5 of culture. Compared with the three-times change, decreasing the change frequency to once did not reduce the capacity to begin maturation (germinal vesicle breakdown of 82 to 86%), to mature (78 to 79%) and to develop into blastocysts after parthenogenetic activation (29 to 32%). Morphological parameters were similar among these treatments. Except for the no medium change treatment, similar colony-forming activity of inner cell mass cells after culturing of blastocysts in leukemia inhibitory factor-containing medium was detected, while the morphology of the colony-forming cells deteriorated in the change-once treatment compared with the change twice or three-times. In conclusion, the efficiency of the preantral follicle culture system could be improved by reducing frequency of medium change up to a 72 h interval (three times in total 216 h culture) without decreasing developmental competence of oocytes.

• Journal of Embryo Transfer
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• v.27 no.2
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• pp.121-126
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• 2012

The objective of this study was to examine the effect of in vitro culture media on embryonic development of in vitro-matured (IVM) oocytes after parthenogenetic activation (PA) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 22~26 h. IVM oocytes were activated by electric pulses and cultured in porcine zygote medium-3 (PZM-3) and North Carolina State University-23 supplemented with essential and non-essential amino acids (NCSU-23aa). These media were further modified by supplementing 2.77 mM myo-inositol, 0.34 mM trisodium citrate, and $10{\mu}M$ ${\beta}$-mercaptoethanol (designated as mPZM-3 and mNCSU-23aa, respectively). Culture of PA embryos in mPZM-3 significantly increased development to the blastocyst stage than culture in NCSU-23aa (36.2% vs. 24.8%, p<0.05). Modified PZM-3 showed a significantly higher blastocyst formation than NCSU-23aa in both groups of embryos that were activated at 44 h and 48 h of IVM (51.0% vs. 35.5% and 49.0% vs. 34.2% in oocytes activated at 44 h and 48 h of IVM, respectively). Irrespective of the follicle diameter where oocytes were collected, embryonic development to the blastocyst stage was increased (p<0.05) by the culture in mPZM-3 compared to culture in NCSU-23aa (25.9% vs. 34.2% and 32.9% vs. 44.8% in embryos derived from small and medium size follicles, respectively). Our results demonstrated that culture media had significant effect on preimplantation development PA embryos and that mPZM-3 was superior to mNCSU-23 in supporting development to the blastocyst stage in pigs. This beneficial effect of mPZM-3 on embryonic development was not impaired by other factors such as time of oocyte activation and origin of immature oocytes (small and medium size follicles).

• Development and Reproduction
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• v.4 no.2
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• pp.175-180
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• 2000

The reproductive cycle of ovoviviparous melania snail, Semisulcospira libeytina libertina was studied in the valley near Naewon Temple in Yangsan, Korea. Water temperature (WT) of the habitat ranged from 1.3$^{\circ}C$ to 22.5$^{\circ}C$ The meat weight rate (MWR) reached the maximum with the beginning of WT increase in March and the minimum in August. Monthly changes in average oocyte diameter showed the maximum (249.6$\pm$2.6 ${\mu}{\textrm}{m}$) in July and the minimum (134.3$\pm$2.8 ${\mu}{\textrm}{m}$) in December S. libertina libertina seemed to be a year-round breeder except for mid-summer and mid-winter. Two main reproductive cycle of the species could be divided into five successive stages: multiplicative (March, October), growing (April and May, November), mature (June and July, December), ovulation (August, January), parturition (September and October, March to May) and resting (September, February) stages in female and multiplicative (March, October), growing (April, November), mature (March to June, December), copulatory (July and August, January), resting (September, February) stages in male.

• Journal of Embryo Transfer
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• v.30 no.4
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• pp.327-333
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• 2015

The objectives of the present study were to select an effective basic medium including its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The ovaries of cows were collected from slaughter house and the follicular fluid was aspirated from 2 to 8 mm diameter follicles. The COCs with more than 3 cumulus cell layers and homogenous cytoplasm were selected for maturation. The oocytes were matured in media for 24 hrs at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The maturation of oocytes was evaluated by examining the presence of first polar body under microscope. An efficient basic medium was determined after culturing COCs in either TCM 199 or SOF medium in Experiment 1. An efficient hormone supplementation was determined after culturing COCs in either FSH or gonadotrophin supplemented TCM 199 in Experiment 2. An efficient protein supplementation was determined after culturing COCs in either FBS or Oestrous cow serum (OCS) supplemented TCM 199 in Experiment 3. The oocyte recovery rate per ovary was 3.35. The overall rate of IVM was 74.6%. The maturation rate was $75.5{\pm}3.9$ and $62.2{\pm}20.2%$ in TCM and SOF medium, respectively (P>0.05). The maturation rate of oocytes was significantly higher ($76.6{\pm}13.2%$) in FSH supplemented medium than gonadotrphin supplemented counterpart ($69.7{\pm}10.8%$) (P<0.05). The maturation rates of oocytes were $81.7{\pm}12.9$ and $85.7{\pm}12.7%$ in medium supplemented with FBS and OCS, respectively (P>0.05). In conclusions, both TCM 199 and SOF supplemented with either FBS or OCS, and FSH may be used as medium for IVM of indigenous zebu oocytes in Bangladesh.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.5
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• pp.385-392
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• 2004

Gonadal development, gametogenesis, reproductive cycle, gonad index, and flesh weight rate of the murex shell (Ocenebra japonica) collected from the rocky intertidal zone of Buan-gun, Jeollabuk-do, Korea were investigated by means of histological method from January to December 2002. O. japonica had separate sexes, and was oviparous. The gonad was widely situated on the surface of the digestive gland located in the rear of the spiral flesh part in the shell. The male penis was located near the two tentacles. The ovary was composed of a number of oogenic follicles, and the testis was composed of several spermatogenic tubules. The size of ripe oocyte was approximately $140{\mu}m$ in diameter. The gonad index (GI) began to increase in March $(33.24{\pm}2.33)$ and reached the maximum in June $(47.77{\pm}1.90)$ Thereafter, the values decreased from July $(45.12{\pm}3.60)$ to October $(19.32{\pm}2.91)$. The flesh weight rate (FWR) began to increase in January $(25.93{\pm}1.32)$ and reached the maxium in May $(31.78{\pm}1.09)$ Thereafter, the values decreased from June $(31.50{\pm}0.66)$ to October $(24.09{\pm}1.60)$. The reproductive cycle could be classified into five successive stages: early active (October to April), late active (January to June), ripe (May to September), spawning (July to September) and recovery (September to February). The reproductive cycle was closely related to the seawater temperature.