P. Vitullo, C. Cossetti, M. A. Virmani^*

Show more
Research, Innovation and Development, Sigma-Tau Health Science International BV, Utrecht, The Netherland.
Show less

Abstract: Introduction: The roles of genetic, epigenetic, metabolic and other environmental factors such as nutrition and stress, are becoming evident for a successful and healthy pregnancy. This raises the possibility and question, if and how, we improve the probability of pregnancy and of a healthy fetus? The present study examined the role of metabolic, antioxidant and minerals and the results suggest that these factors may positively influence the oocyte quality and the pregnancy rate. Methods: CD1 female mice aged 15 and 5 weeks were divided into four groups of ten each and treated by intragastric gavage daily for 3 weeks. G1: Vehicle; G2: Carnitines (L-carnitine 0.4 mg and acetyl-L-carnitine 0.12 mg/mouse); G3: Microelements (Zinc 4 ng, Copper 0.8 ng, Iron 7 ng/mouse); G4: G3+G2. At the end of the treatment period superovulation was induced and oocytes were collected to assess their quantity and quality. Further, in vitro fertilization (IVF) experiments were performed to assess the preimplantation embryo development. The birth success rate was also analyzed in old and young female. The mice were in vivo fertilized. qRT-PCR were performed to analyze a possible modulation in key genes of the reproductive process. Results: The number of oocytes was significantly higher in groups 2 and 4 compared to the control group. The oocyte number in group 3 was not affected. The level of degraded oocytes was 29.1% and 19.3% (group 2 and 4) versus 34.3% (control). Concomitantly, the numbers of embryos arriving to successful birth were also increased in G4, both in the old and young group of mice. Preliminary analysis of genes affected evidenced that AMH was up regulated in the ovary and KITL in the uterus in group 2. Conclusion: Results showed that L-carnitine, acetyl-L-carnitine and micronutrients were able to improve both oocytes quality and success rate of pregnancy. Further studies are planned to further examine ways to improve pregnancy and fetal health.

Keywords: Pregnancy, L-Carnitine, Acetyl-L-Carnitine, Micronutrients, Oocytes, Birth, Fetal Health, Embryos, Key Genes, Fertilization, Reproductive

Cite this paper: Vitullo, P. , Cossetti, C. and Virmani, M. (2018) Effect of Specific Nutrients on Ovulation, Oocytes Development, Gene Expression and Coupling Success in Mice. International Journal of Clinical Medicine, 9, 660-674. doi: 10.4236/ijcm.2018.99055.

References

[1]   Hakim, R.B., Gray, R.H. and Zacur, H. (1998) Alcohol and Caffeine Consumption and Decrease Fertility. Fertility and Sterility, 70, 632-637.
https://doi.org/10.1016/S0015-0282(98)00257-X

[2]   Gray, R.H. and Becker, S. (2000) Selected Topics in the Epidemiology of Reproductive Outcomes. Epidemiologic Reviews, 22, 71-75.
https://doi.org/10.1093/oxfordjournals.epirev.a018027

[3]   Virmani, M.A., et al. (2013) Food, Nutrigenomics and Neurodegeneration—Neuro-protection by What You Eat! Molecular Neurobiology, 48, 353-362.
https://doi.org/10.1007/s12035-013-8498-3

[4]   Virmani, M.A. and Angeli, A. (2013) Role of Nutrigenomics and Metabolic Markers in Evaluating Oxidative Stress and Inflammation Dysfunctions in Female Fertility. Abstract and Posters, Vitafoods, Geneva.

[5]   Kim, C.S., et al. (1990) L-Carnitine Prevents Mitochondrial Damage Induced by Octanoid Acid in the Rat Choroid Plexus. Brain Research, 536, 335-338.
https://doi.org/10.1016/0006-8993(90)90046-E

[6]   Virmani, M.A., et al. (1995) Protective Actions of L-Carnitine and Acetyl-L-Carnitine on the Neurotoxicity Evoked by Mitochondrial Uncoupling or Inhibitors. Pharmacological Research, 32, 383-389.
https://doi.org/10.1016/S1043-6618(05)80044-1

[7]   Binienda, Z., et al. (1999) Protective Effect of L-Carnitine in the Neurotoxicity Induced by Mitochondrial Inhibitor 3-Nitrorpopionic Acid (3-NPA). Annals of the New York Academy of Sciences, 890, 173-178.
https://doi.org/10.1111/j.1749-6632.1999.tb07992.x

[8]   Matzuk, M.M. and Lamb, D.J. (2002) Genetic Dissection of Mammalian Fertility Pathways. Nature Cell Biology, 4, s41-49.
https://doi.org/10.1038/ncb-nm-fertilityS41

[9]   Virant-Klun, I., Knez, K., Tomazevic, T. and Skutella, T. (2013) Gene Expression Profiling of Human Oocytes Developed and Matured In Vivo or In Vitro. BioMed Research International, 2013, Article ID: 879489.
https://doi.org/10.1155/2013/879489

[10]   Mansour, G., Abdelrazik, H., Sharma, R.K., Radwan, E., Falcone, T. and Agarwal, A. (2009) L-Carnitine Supplementation Reduces Oocyte Cytoskeleton Damage and Embryo Apoptosis Induced by Incubation in Peritoneal Fluid from Patients with Endometriosis. Fertility and Sterility, 91, 2079-2086.
https://doi.org/10.1016/j.fertnstert.2008.02.097

[11]   Virmani, M.A., Krismanovic, L.Z., Stojilkovic, S.S. and Catt, K.J. (1991) Stimulatory Effects of L-Acetylcarnitine on the Pituitary-Gonadal Axis in Female Rats. In: Adashi, E.Y. and Mancus, S., Eds., Major Advances in Human Female Reproduction, Raven Press, 73, 291-296.

[12]   Debbie, M., Frida, E., Isabelle, L., Stephanie, B., Timur, G. and Yves, M. (2012) Carnitine Content in the Follicular Fluid and Expression of the Enzymes Involved in Beta-Oxidation in Oocyte and Cumulus Cells. Journal of Assisted Reproduction and Genetics, 29, 1221-1225.
https://doi.org/10.1007/s10815-012-9855-2

[13]   Dunning, K.R., Cashman, K., Russel, D.L., Thompson, J.G., Norman, R.J. and Robker, R.L. (2010) Beta-Oxidation Is Essential for Mouse Oocyte Development Competence and Early Embryo Development. Biology of Reproduction, 83, 909-918.
https://doi.org/10.1095/biolreprod.110.084145

[14]   Virmani, M.A., Zerelli, S., Vitullo, P. and Cossetti, C. (2015) Effect of Nutrients on Ovulation and Oocytes Quality in Mice. Giornale Italiano di Ostetricia e Ginecologia, 37, 212-214.

[15]   Virmani, A., Diedenhofen, A. and Zerelli, S. (2014) Mitochondriotropic Compounds in Energy, Oxidative Stress and Inflammation: Role in Reproductive Health, Fertility and Successful Pregnancy. Giornale Italiano di Ostetricia e Ginecologia, 36, 293-296.

[16]   Vitullo, P., Sciamanna, I., Baiocchi, M., Sinibaldi-Vallebona, P. and Spadafora, C. (2012) LINE-1 Retrotransposon Copies Are Amplified during Murine Early Embryo Development. Molecular Reproduction and Development, 79, 118-127.
https://doi.org/10.1002/mrd.22003

[17]   Meng, R. (2018) Relative Quantification: Data Management and Analysis Settings. Bio-Rad Laboratories Gene Expression Division.
http://www.bio-rad.com/genomics/pcrsupport

[18]   Greene, A.D., Patounakis, G. and Segars, J.H. (2014) Genetic Associations with Diminished Ovarian Reserve: A Systematic Review of the Literature. Journal of Assisted Reproduction and Genetics, 31, 935-946.
https://doi.org/10.1007/s10815-014-0257-5

[19]   Yding Andersen, C. (2017) Inhibin-B Secretion and FSH Isoform Distribution May Play an Integral Part of Follicular Selection in the Natural Menstrual Cycle. Molecular Human Reproduction, 23, 16-24.
https://doi.org/10.1093/molehr/gaw070

[20]   Beck, F., Erler, T., Russell, A. and James, R. (1995) Expression of Cdx-2 in the Mouse Embryo and Placenta: Possible Role in Patterning of the Extra-Embryonic Membranes. Developmental Dynamics, 204, 219-227.
https://doi.org/10.1002/aja.1002040302

[21]   Strumpf, D., Mao, C.-A., Yamanaka, Y., Ralston, A., Chawengsaksophak, K., Beck, F. and Rossant, J. (2005) Cdx2 Is Required for Correct Cell Fate Specification and Differentiation of Trophectoderm in the Mouse Blastocyst. Development, 132, 2093-2102.
https://doi.org/10.1242/dev.01801