• Title/Summary/Keyword: Canine oocyte

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Morphological Assessment of Ovulated and In Vitro Immature Canine Oocytes and Biological Availability according to the Size at Different Reproductive Stages

  • Hossein Mohammad-Shamim;Kim Min-Kyu;Jang Goo;Fibrianto Yuda-Heru;Oh Huyn-Joo;Kim Hye-Jin;Kim Joung-Joo;Kang Sung-Keun;Lee Byeong-Chun
    • Reproductive and Developmental Biology
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    • v.30 no.2
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    • pp.99-105
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    • 2006
  • The growing oocytes become progressively capable of resuming meiosis, and full meiotic competence appear when they are about 80% of the size of fully grown oocytes. As hormonal influences vary at different stages of reproductive cycle, the size of oocytes may vary according to the reproductive stages. The present study was designed to compare the diameter between the ovulated and freshly collected immature canine oocytes. The ovulated oocytes were collected 72 hr after ovulation by oviductal tube flushing by laparotomy under general anesthesia. Immature oocytes were collected by ovarian slicing method. Diameter of all oocytes was measured directly using epiflurescence microscope with a calibrated micro-eyepiece micrometer at ${\times}200$ magnification. The thickness of zona pellucida and diameter of cytoplasm were measured separately and recorded. A total of 2209 zona intact oocytes were collected, among them 628 from anestrus, 675 from follicular, 838 from luteal and 68 by fallopian tubes flushing methods. The average number of oocytes was 104.7, 168.8, 119.7 and 11.3 for anestrus, follicular, luteal and fallopian tubes flushing methods, respectively. The average diameters of the ooplasm and oocyte were significantly varied in different reproductive stages as well as with ovulated oocytes (P<0.05). The average diameter of ooplasm and oocyte was 115.6 and 127.7, 143.0 and 162.0, 134.6 and 150.6, 159.6 and 185.6 for anestrus, follicular, luteal and ovulated oocytes, respectively. Highest number of oocytes with larger diameter could be collected from the follicular and luteal stages. In conclusion, the follicular and luteal ovaries are the best sources of oocytes for canine IVM.

Effect of Various Caffeine Concentrations and Fertilization Time in In Vitro Fertilization of Canine Oocytes

  • Kim, Bong-Soo;Yoo, Dong-Hoon;Shin, Mi-Jung;Kim, Hae-Jung;Lee, Dong-Seok;Hyun, Byung-Hwa;Lee, Sang-Gyu;Park, Young-Sik;Ha, Ji-Hong;Ryoo, Zae-Young
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.217-222
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    • 2008
  • The techniques of IVM, IVF and IVC of canine oocytes may provide useful information for gamete salvage programs and the conservation of endangered canidae. This investigation has been made to determine the efficiency of in vitro maturation (IVM) as a basic experiment to study the development of canine oocytes after in vitro fertilization (IVF). The rate of oocytes developing to the Mil stage was higher in the hormone treated group (10 IU/ml hCG+eCG, 14.7%, p<0.05) than in the control group (0 IU/ml hCG+eCG, 10.0%). The monospermy and pronuclear rates of canine oocytes were investigated after caffeine treatment on IVF. Canine oocytes were fertilized in the Fert-TALP medium supplemented with 0, 10, 20 or 30 mM caffeine (Fert I, Fert II, Fert III or Fert IV, respectively). The highest pronuclear formation rate was obtained in the Fert I for 24 h IVF (6.7%, 6/89). Therefore, it is believed that unlike in other mammals, caffeine in canine IVF does not increase the efficiency of fertilization rate, and is not an important factor.

Effects of Hormone and Na-Pyruvate on the In Vitro Maturation of Canine Oocytes (개 난자의 체외성숙에 미치는 호르몬과 Na-Pyruvate의 영향)

  • Kim Cheon-Ho
    • Reproductive and Developmental Biology
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    • v.30 no.1
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    • pp.7-11
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    • 2006
  • This study was conducted to examine the effects of hormone and sodium pyruvate on in vitro maturation of canine oocytes. Canine oocytes were collected from the ovaries of dogs and cultured in NCSU-37 medium with hormones and sodium pyruvate for 72 hr. Oocytes matured to the metaphase II (MII) stage were observed only from estradiol $17{\beta}\;(E_2)$, and the presence of gonadotropin did not improve the nuclear maturation. No oocytes were developed to the MII stage when $E_2$ was added to medium during the first 6 and 24 hrs of culture period. The presence of $E_2$ during the whole culture period enhanced the nuclear maturation to the MII stage (6.0%, P<0.05). High concentration of sodium pyruvate (2.5 mM) slightly enhanced the nuclear maturation to the metapahse I (HMI) stage, but not the MII stage. the result of the present study shows that the presence of $ E_2$ during the whole culture period of 72 hr enhances the maturation of canine oocytes to the M stage, but sodium pyruvate does not affect the nuclear maturation of the canine oocytes.

Interspecies Somatic Cell Nuclear Transfer Technique for Researching Dog Cloning and Embryonic Stem Cells

  • Sugimura, Satoshi;Sato, Eimei
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.1-8
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    • 2011
  • Large quantities of high-quality recipient oocytes with uniform cytoplasm are needed for research in the promising field of somatic cell nuclear transfer (SCNT) and embryonic stem cell research. In canines, however, it is difficult to obtain large quantities of oocytes because each donor produces a limited number of mature oocytes in vivo. Although in vitro maturation (IVM) is considered an alternative approach to oocyte production, this technique is still too rudimentary to be used for the production of highquality, uniform oocytes in large quantities. One technique for overcoming this difficulty is to use oocytes obtained from different species. This technique is known as interspecies SCNT (iSCNT). This review provides an overview of recent advances in canine - porcine interspecies SCNT.

발정 주기에 따른 개 미성숙 난자의 핵발달단계의 비교

  • 이효상;윤희준;이영호;전세진;서영일;조수진;최유진;조성균;공일근
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.241-241
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    • 2004
  • 본 연구는 발정주기에 따른 개의 미성숙난자의 체외배양 0시간째 핵의 발달단계를 비교ㆍ검토하였다. 발정주기가 다른 개의 난소를 적출 하여 항생제가 첨가된 38℃의 0.9% 생리식염수가 들어있는 보온병에 넣어 2시간 이내에 연구실로 운반하고 난소를 면도날로 세절하여 난포란을 회수하였고, 난구세포 제거 후 Chohan과 Hunter (2003) 방법에 준하여 10 ug/ml Hoechst 33342를 이용한 핵염색을 실시하여 핵의 모양을 판명하였다. 조사된 결과는 SAS 8.0 Package를 이용하여 통계분석을 실시하였다. (중략)

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