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volume 47 issue 3 (june 2013) : 212-219

EFFECTS OF DIFFERENT MITOTIC INDUCERS ON MULTIPLICATION OF PARTHENOGENETIC EMBRYONIC CELLS IN BUFFALO

B
Bijay Kumar Chaudhari*
B
B.C. Das
A
A.K. Singh1
J
J.K. Singh2
P
P. K. Maurya1
S
S. Kumar1
1Division of Physiology & Climatology Indian Veterinary Research Institute, Izatnagar - 243 122, India

  • Submitted|

  • First Online |

  • doi

Cite article:- Chaudhari* Kumar Bijay, Das B.C., Singh1 A.K., Singh2 J.K., Maurya1 K. P., Kumar1 S. (2025). EFFECTS OF DIFFERENT MITOTIC INDUCERS ON MULTIPLICATION OF PARTHENOGENETIC EMBRYONIC CELLS IN BUFFALO. Indian Journal of Animal Research. 47(3): 212-219. doi: .

ABSTRACT

The present study was conducted to see the effects of mitotic inducers on multiplication of cultured parthenogenetic blastomere cells in buffalo. Total 1364 ovaries were aspirated and out of which 1034 good quality oocytes were recovered at the rate of 75.80 %. The overall recovery rate of oocytes was 0.75 / ovary in the present study. In protocol-I, activation of matured oocytes was done with 5% ethanol for 7min+6-DMAP for 4hrs where as in protocol-II; activation was done with with 5µm ionomycin for 5min+6DMAP for 4hrs.  In protocol I, the cleavage rate and morula development was 60.47% and 10.23% respectively, where as in protocol-II, the cleavage rate and morula development was 38.11% and 7.69% respectively. It was observed that protocol-I was found to be effective and it differs from protocol II in cleavage rate as well morula development.  Blastomeres were cultured for day 0, 4, 8, 12 and day 20 on feeder layers in media containing different factors like mitotic inducers (melatonin and lipopolysaccharides) and retinoic acid blocker (citral). It was observed that the colony size was comparable to colony size of LPS (lipopolysaccharides) on different days of culture. The colony size developed in presence of melatonin was comparatively larger as compared to any other culture condition for all days of culture.

REFERENCES

  1. Arici, A. Engin, O. Attar, E. and Olive, D.L. (1995). Modulation of leukemia inhibitory factor gene expression and protein biosynthesis in human endometrium. J. Clin. Endocrinol. Metab. 80: 1908–1915.
  2. Boediono and Suzuki. (1994). Offsprig born from chimeras reconstructed from parthenogenetic and in vitro fertilized bovine embryo. Mol. Reproduction and Development. 53(2): 159 70.
  3. De Luca, L. M. (1991). Retinoids and their receptors in differentiation, embryogenesis, and neoplasia. FASEB J. 5: 2924-2933.
  4. Green, J. B. (1990). Retinoic acid: the morphogen of the main body axis? Bioessays. 12: 437-439.
  5. Gupta, M. K. Sang, J. U. and Lee, H. K. (2007). Embryo development, embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin. Mol.Reprod. Dev. 74 (4):435-44.
  6. Harms,C. M. Lautenschlager, A. Bergk, D. Freyer, M. Weih, U.Dirnagl, J.R. Weber and Hortnagl, H. (2000). Melatonin is protective in necrotic but not in caspase-dependent, free radical-independent apoptotic neuronal cell death in primary neuronal cultures. FASEB J. 14: 1814–1824
  7. Hidalgo, C. Diez, C. Duque, P. Prendes, J.M. Rodriguez, A. Goyache, F. Fernandez, I. Facal, N. Ikeda, S. and Alonso- Montes, C. (2005). Oocytes recovered from cows treated with retinol become unviable as blastocysts produced in vitro. Reproduction. 129:411–421
  8. Jones, K. T. Carroll, J. and Whittingham, D. G. (1995). Ionomycin, Thapsigargin, Ryanodine, and Sperm induced Ca2+ release increase during meiotic maturation of mouse oocytes. J. Biol. Chem. 270: 6671–6677.
  9. Kauma, S.W. (2000). Cytokines in implantation. J. Reprod. Fertil. Suppl. 55:31-42.
  10. Kim, M. J. Kim, H. K. Kim, B. S. and Yim, S. V. (2004). Melatonin increases cell proliferation in the dentate gyrus of maternally separated rats. J. Pineal Res. 37: 193–197.
  11. Kline, D. and Kline, J. T. (1992). Repetitive calcium transients and the role of calcium in exocytosis mouse egg and suppresses repetitive calcium transients in the fertilized egg. J. Biol. Chem. 267: 17624–17630.
  12. Kong, X. Li, X. Cai, Z. Yang, N. Liu Y. and Shu. J. (2008). Melatonin regulates the viability and differentiation of rat midbrain neural stem cells. Cell Mol. Neurobiol. 28: 569–579.
  13. Kronmiller, J. E. Beeman, C. S. Nguyen T. and Berndt, W. (1995a). Blockade of the initiation of murine odontogenesis in vitro by citral, an inhibitor of endogenous retinoic acid synthesis. Archs oral Biol. 40: 645-652.
  14. Lawrence, J.L. Payton, R.R. Godkin, J.D. Saxton, A.M. Schrick, F.N. and Edwards, J.L. (2004). Retinol improves development of bovine oocytes compromised by heat stress during maturation. J. Dairy Sci. 87:2449–2454.
  15. Leid, M. Kastner, P. Lyons, R. Nakshatri, H. Saunders, M. Zacharewski, T. Chen, J. Y. Staub, A. Garnier, J. M. and Mader, S. (1992). Puriûcation, cloning, and RXR identity of the HeLa cell factor with which RAR or TR heterodimerizes to bind target sequences efficiently. Cell. 68:377-395.
  16. Leroy, J.L. Genicot, G. Donnay, I. and Van Soom, A. (2005). Evaluation of the lipid content in bovine oocytes and embryos with Nile red: a practical approach. Reprod. Domest. Anim. 40:76–78.
  17. Livingston, T. Eberhardt, D. Edwards, J.L. and Godkin, J. (2004). Retinol improves bovine embryonic development in vitro. Reprod. Biol. Endocrinol. 2:83.
  18. Moriya, T. Horie, N. Mitome, M. and Shinohara, K. (2007). Melatonin influences the proliferative and differentiative activity of neural stem cells. J. Pineal Res. 42: 411–418.
  19. Nagai, T. (1987). Parthenogenetic activation of cattle follicular oocytes in vitro with ethanol. Gamete. Res. 16: 243- 249.
  20. Reiter, R. J. (1998b). Oxidative damage in the central nervous system: protection by melatonin. Prog. Neurobiol. 56: 359–384.
  21. Rodrýguez G. R. Klempin, F. Babu, H. King, G. B. and Kempermann, G. (2009). Melatonin Modulates Cell Survival of New Neurons in the Hippocampus of Adult Mice. Neuropsychopharmacology. 34: 2180–2191.
  22. Ross, C. (1975). Alternative Livestock: with particular response to the water buffalo (Bubalus bubalis). Meat. 1:507- 524.
  23. Schuh, T. Hall, B. Kraft, C. J. Privalsky, M. and Kimmelman, D. (1993). V-erb and citral reduce tertagenic effects of all-trans-retinoic acid and retinol, respectively,in Xenopus embryogenesis. Dev. 119: 785-798.
  24. Turunen, O. Lundqvist, C. and de la Chapelle, A. (1977). Stimulation of human fetal lymphocytes by lipopolysaccharide B in culture. Scand. J. Immunol. 6(4):335-42.
  25. Vijayalaxmi, Reiter, R. J. Leal B. Z. and Meltz, M. L. (1996). Effect of melatonin on mitotic and proliferation indices, and sister chromatid exchange in human blood lymphocytes. Mutat. Res. 351:187-91.
  26. Wang, Z.G. Wang, W. Yu, S.D. and Xu, Z. R. (2007). Effect of different activation protocols on preimplantation development and apoptosis and ploidy of bovine parthenogenetic embryos. Anim. Reprod Sci. 105: 292-301.
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Indian Journal of Animal Research
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    volume 47 issue 3 (june 2013) : 212-219

    EFFECTS OF DIFFERENT MITOTIC INDUCERS ON MULTIPLICATION OF PARTHENOGENETIC EMBRYONIC CELLS IN BUFFALO

    B
    Bijay Kumar Chaudhari*
    B
    B.C. Das
    A
    A.K. Singh1
    J
    J.K. Singh2
    P
    P. K. Maurya1
    S
    S. Kumar1
    1Division of Physiology & Climatology Indian Veterinary Research Institute, Izatnagar - 243 122, India

    • Submitted|

    • First Online |

    • doi

    Cite article:- Chaudhari* Kumar Bijay, Das B.C., Singh1 A.K., Singh2 J.K., Maurya1 K. P., Kumar1 S. (2025). EFFECTS OF DIFFERENT MITOTIC INDUCERS ON MULTIPLICATION OF PARTHENOGENETIC EMBRYONIC CELLS IN BUFFALO. Indian Journal of Animal Research. 47(3): 212-219. doi: .

    ABSTRACT

    The present study was conducted to see the effects of mitotic inducers on multiplication of cultured parthenogenetic blastomere cells in buffalo. Total 1364 ovaries were aspirated and out of which 1034 good quality oocytes were recovered at the rate of 75.80 %. The overall recovery rate of oocytes was 0.75 / ovary in the present study. In protocol-I, activation of matured oocytes was done with 5% ethanol for 7min+6-DMAP for 4hrs where as in protocol-II; activation was done with with 5µm ionomycin for 5min+6DMAP for 4hrs.  In protocol I, the cleavage rate and morula development was 60.47% and 10.23% respectively, where as in protocol-II, the cleavage rate and morula development was 38.11% and 7.69% respectively. It was observed that protocol-I was found to be effective and it differs from protocol II in cleavage rate as well morula development.  Blastomeres were cultured for day 0, 4, 8, 12 and day 20 on feeder layers in media containing different factors like mitotic inducers (melatonin and lipopolysaccharides) and retinoic acid blocker (citral). It was observed that the colony size was comparable to colony size of LPS (lipopolysaccharides) on different days of culture. The colony size developed in presence of melatonin was comparatively larger as compared to any other culture condition for all days of culture.

    REFERENCES

    1. Arici, A. Engin, O. Attar, E. and Olive, D.L. (1995). Modulation of leukemia inhibitory factor gene expression and protein biosynthesis in human endometrium. J. Clin. Endocrinol. Metab. 80: 1908–1915.
    2. Boediono and Suzuki. (1994). Offsprig born from chimeras reconstructed from parthenogenetic and in vitro fertilized bovine embryo. Mol. Reproduction and Development. 53(2): 159 70.
    3. De Luca, L. M. (1991). Retinoids and their receptors in differentiation, embryogenesis, and neoplasia. FASEB J. 5: 2924-2933.
    4. Green, J. B. (1990). Retinoic acid: the morphogen of the main body axis? Bioessays. 12: 437-439.
    5. Gupta, M. K. Sang, J. U. and Lee, H. K. (2007). Embryo development, embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin. Mol.Reprod. Dev. 74 (4):435-44.
    6. Harms,C. M. Lautenschlager, A. Bergk, D. Freyer, M. Weih, U.Dirnagl, J.R. Weber and Hortnagl, H. (2000). Melatonin is protective in necrotic but not in caspase-dependent, free radical-independent apoptotic neuronal cell death in primary neuronal cultures. FASEB J. 14: 1814–1824
    7. Hidalgo, C. Diez, C. Duque, P. Prendes, J.M. Rodriguez, A. Goyache, F. Fernandez, I. Facal, N. Ikeda, S. and Alonso- Montes, C. (2005). Oocytes recovered from cows treated with retinol become unviable as blastocysts produced in vitro. Reproduction. 129:411–421
    8. Jones, K. T. Carroll, J. and Whittingham, D. G. (1995). Ionomycin, Thapsigargin, Ryanodine, and Sperm induced Ca2+ release increase during meiotic maturation of mouse oocytes. J. Biol. Chem. 270: 6671–6677.
    9. Kauma, S.W. (2000). Cytokines in implantation. J. Reprod. Fertil. Suppl. 55:31-42.
    10. Kim, M. J. Kim, H. K. Kim, B. S. and Yim, S. V. (2004). Melatonin increases cell proliferation in the dentate gyrus of maternally separated rats. J. Pineal Res. 37: 193–197.
    11. Kline, D. and Kline, J. T. (1992). Repetitive calcium transients and the role of calcium in exocytosis mouse egg and suppresses repetitive calcium transients in the fertilized egg. J. Biol. Chem. 267: 17624–17630.
    12. Kong, X. Li, X. Cai, Z. Yang, N. Liu Y. and Shu. J. (2008). Melatonin regulates the viability and differentiation of rat midbrain neural stem cells. Cell Mol. Neurobiol. 28: 569–579.
    13. Kronmiller, J. E. Beeman, C. S. Nguyen T. and Berndt, W. (1995a). Blockade of the initiation of murine odontogenesis in vitro by citral, an inhibitor of endogenous retinoic acid synthesis. Archs oral Biol. 40: 645-652.
    14. Lawrence, J.L. Payton, R.R. Godkin, J.D. Saxton, A.M. Schrick, F.N. and Edwards, J.L. (2004). Retinol improves development of bovine oocytes compromised by heat stress during maturation. J. Dairy Sci. 87:2449–2454.
    15. Leid, M. Kastner, P. Lyons, R. Nakshatri, H. Saunders, M. Zacharewski, T. Chen, J. Y. Staub, A. Garnier, J. M. and Mader, S. (1992). Puriûcation, cloning, and RXR identity of the HeLa cell factor with which RAR or TR heterodimerizes to bind target sequences efficiently. Cell. 68:377-395.
    16. Leroy, J.L. Genicot, G. Donnay, I. and Van Soom, A. (2005). Evaluation of the lipid content in bovine oocytes and embryos with Nile red: a practical approach. Reprod. Domest. Anim. 40:76–78.
    17. Livingston, T. Eberhardt, D. Edwards, J.L. and Godkin, J. (2004). Retinol improves bovine embryonic development in vitro. Reprod. Biol. Endocrinol. 2:83.
    18. Moriya, T. Horie, N. Mitome, M. and Shinohara, K. (2007). Melatonin influences the proliferative and differentiative activity of neural stem cells. J. Pineal Res. 42: 411–418.
    19. Nagai, T. (1987). Parthenogenetic activation of cattle follicular oocytes in vitro with ethanol. Gamete. Res. 16: 243- 249.
    20. Reiter, R. J. (1998b). Oxidative damage in the central nervous system: protection by melatonin. Prog. Neurobiol. 56: 359–384.
    21. Rodrýguez G. R. Klempin, F. Babu, H. King, G. B. and Kempermann, G. (2009). Melatonin Modulates Cell Survival of New Neurons in the Hippocampus of Adult Mice. Neuropsychopharmacology. 34: 2180–2191.
    22. Ross, C. (1975). Alternative Livestock: with particular response to the water buffalo (Bubalus bubalis). Meat. 1:507- 524.
    23. Schuh, T. Hall, B. Kraft, C. J. Privalsky, M. and Kimmelman, D. (1993). V-erb and citral reduce tertagenic effects of all-trans-retinoic acid and retinol, respectively,in Xenopus embryogenesis. Dev. 119: 785-798.
    24. Turunen, O. Lundqvist, C. and de la Chapelle, A. (1977). Stimulation of human fetal lymphocytes by lipopolysaccharide B in culture. Scand. J. Immunol. 6(4):335-42.
    25. Vijayalaxmi, Reiter, R. J. Leal B. Z. and Meltz, M. L. (1996). Effect of melatonin on mitotic and proliferation indices, and sister chromatid exchange in human blood lymphocytes. Mutat. Res. 351:187-91.
    26. Wang, Z.G. Wang, W. Yu, S.D. and Xu, Z. R. (2007). Effect of different activation protocols on preimplantation development and apoptosis and ploidy of bovine parthenogenetic embryos. Anim. Reprod Sci. 105: 292-301.
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