Effects of Nicotine and Tocotrienol-Rich Fraction Supplementation on Cytoskeletal Structures of Murine Pre-Implantation Embryos

Nurul Kamsani Hamirah, Yuhaniza Shafinie Kamsani, Nor-Ashikin Mohamed Nor Khan, Sharaniza Ab Rahim, Mohd Hamim Rajikin, Nurul Kamsani Hamirah, Yuhaniza Shafinie Kamsani, Nor-Ashikin Mohamed Nor Khan, Sharaniza Ab Rahim, Mohd Hamim Rajikin

Abstract

BACKGROUND Cytoskeletal structures, in particular actin and tubulin, provide a fundamental framework in all cells, including embryos. The objective of this study was to evaluate the effects of nicotine, which is a source of oxidative stress, and subsequent supplementation with Tocotrienol-rich fraction (TRF) on actin and tubulin of 2- and 8-cell murine embryos. MATERIAL AND METHODS Thirty female Balb/C mice were divided into 4 groups: Group 1 received: subcutaneous (sc) injection of 0.9% NaCl; Group 2 received sc injection of 3.0 nicotine mg/kg bw/day; Group 3 received 3.0 sc injection of nicotine mg/kg bw/day +60 mg/kg bw/day TRF; and Group 4 received 60 sc injection of TRF mg/kg bw/day for 7 consecutive days. The animals were superovulated with 5 IU PMSG followed by 5 IU hCG 48 h later. Animals were cohabited with fertile males overnight and euthanized through cervical dislocation at 24 h post coitum. Embryos at the 2- and 8-cell stages were harvested, fixed, and stained to visualize actin and tubulin distributions by using CLSM. RESULTS Results showed that at 2-cell stage, actin intensities were significantly reduced in the nicotine group compared to that of the control group (p<0.001). In Group 3, the intensity of actin significantly increased compared to that of the nicotine group (p<0.001). At 8-cell stage, actin intensity of the nicotine group was significantly lower than that of the control group (p<0.001). The intensities of actin in Group 3 were increased compared to that of nicotine treatment alone (p<0.001). The same trend was seen in tubulin at 2- and 8-cell stages. Interestingly, both actin and tubulin structures in the TRF-treated groups were enhanced compared to the control. CONCLUSIONS This study suggests that TRF prevents the deleterious effects of nicotine on the cytoskeletal structures of 2- and 8-cell stages of pre-implantation mice embryos in vitro.

Figures

Figure 1
Figure 1
Immunofluorescent intensities of actin at 2-cell (A–D) and 8-cell (E–H) stages of embryos. Control group (A, E), nicotine group (B, F), nicotine + TRF group (C, G) and TRF group (D, H) embryos stained with Alexa Fluor 635 phallodin for actin (red) were observed under a CLSM.
Figure 2
Figure 2
Comparisons of actin intensity (%) between all treatment groups. Values with different superscripts are significantly different. Nic – Nicotine, TRF – tocotrienol-rich.
Figure 3
Figure 3
Immunofluorescent intensities of tubulin at 2-cell (A–D) and 8-cell (E–H) stages of embryos. Control group (A, E), nicotine group (B, F), nicotine + TRF group (C, G) and TRF group (D, H) embryos stained with anti-α-tubulin for tubulin (green) were observed under a CLSM.
Figure 4
Figure 4
Comparisons of tubulin intensity (%) between all treatment groups. Values with different superscripts are significantly different. Nic – Nicotine, TRF – tocotrienol-rich.

References

    1. Ko TJ, Tsai LY, Chu LC, et al. Parental smoking during pregnancy and its association with low birth weight, small for gestational age, and preterm birth offspring: A birth cohort study. Pediatr Neonatol. 2014;55:20–27.
    1. Harrod CS, Reynolds RM, Chasan-Taber L, et al. Quantity and timing of maternal prenatal smoking on neonatal body composition: the healthy start study. J Pediatr. 2014;165:707–12.
    1. Blatt K, Moore E, Chen A, et al. Association of reported trimester-specific smoking cessation with fetal growth restriction. Obstet Gynecol. 2015;125:1452–59.
    1. Lintsen A, Pasker-de Jong P, De Boer E, et al. Effects of subfertility cause, smoking and body weight on the success rate of IVF. Hum Reprod. 2005;20(7):1867–75.
    1. Thielen A, Klus H, Müller L. Tobacco smoke: unraveling a controversial subject. Exp Toxicol Pathol. 2008;60:141–56.
    1. Knopik VS, Maccani MA, Francazio S, McGeary JE. The epigenetics of maternal cigarette smoking during pregnancy and effects in child development. Dev Psychopathol. 2012;24:1377–90.
    1. Lee KW, Richmond R, Hu P, et al. Prenatal exposure to maternal cigarette smoking and DNA Methylation: Epigenome-wide association in a discovery sample of adolescents and replication in an independent cohort at birth through 17 years of age. Environ Health Perspect. 2015;123:193–99.
    1. Pirini F, Guida E, Lawson F, et al. Nuclear and mitochondrial DNA alterations in newborns with prenatal exposure to cigarette smoke. Int J Environ Res Public Health. 2015;12:1135–55.
    1. Schneider S, Huy C, Schütz J, Diehl K. Smoking cessation during pregnancy: A systematic literature review. Drug Alcohol Rev. 2010;29:81–90.
    1. Lisboa PC, de Oliveira E, de Moura EG. Obesity and endocrine dysfunction programmed by maternal smoking in pregnancy and lactation. Front Physiol. 2012;3:437–46.
    1. Ornoy A. Embryonic oxidative stress as a mechanism of teratogenesis with special emphasis on diabetic embryopathy. Reprod Toxicol. 2007;24:31–41.
    1. Bellomo G, Mirabelli F. Oxidative stress and the cytoskeletal alterations. Ann NY Acad Sci. 1992;663:97–109.
    1. Simonian NA, Coyle JT. Oxidative stress in neurodegenerative diseases. Annu Rev Pharmacol Toxicol. 1996;36:83–106.
    1. Wang X, Falcone T, Attaran M, et al. Vitamin C and Vitamin E supplementation reduce oxidative stress-induced embryo toxicity and improve the blastocysts development rate. Fert Steril. 2002;78:1271–77.
    1. Rajikin MH, Latif ES, Mar MR, et al. Deleterious effects of nicotine on the ultrastructure of oocytes: Role of gamma tocotrienol. Med Sci Monit. 2009;15(12):BR378–83.
    1. Barnett DK, Clayton MK, Kimura J, Bavister BD. Glucose and phosphate toxicity in hamster preimplantation embryos involves disruption of cellular organization, including distribution of active mitochondria. Mol Reprod Dev. 1997;48:227–37.
    1. Boldogh I, Vojtov N, Karmon S, Pon LA. Interaction between mitochondria and the actin cytoskeleton in budding yeast requires two integral mitochondrial outer membrane proteins, Mmm1p and Mdm10p. J Cell Biol. 1998;141:1371–81.
    1. Valderrama F, Babia T, Ayala I, et al. Actin microfilaments are essential for the cytological position and morphology of the golgi complex. Eu J Cell Biol. 1998;76:9–17.
    1. Bihan TL, Pelletier D, Tancrede P, et al. Effect of polar headgroup of phospholipids on their interaction with actin. J Colloid and Interface Sci. 2005;288:88–96.
    1. Charras GT, Yarrow JC, Horton MA, et al. Non-equilibration of hydrostatic pressure in blebbing cells. Nature. 2005;435:365–69.
    1. Doherty GJ, McMahon HT. Mediation, modulation and consequences of membrane-cytoskeleton interactions. Annu Rev Biophys. 2008;37:65–95.
    1. Kamsani YS, Rajikin MH, Chatterjee A, et al. Impairment of in vitro embryonic development with a corresponding elevation of oxidative stress following nicotine treatment in mice: Effect of variation in treatment duration. Biomed Res. 2010;21:359–64.
    1. Nesaretnam K, Meganatha P, Veerasenan SD, Selvaduray KR. Tocotrienols and breast cancer: The evidence to date. Genes Nutr. 2012;7:3–9.
    1. Yuen KH, Wong JW, Lim AB, et al. Effect of mixed-tocotrienols in hypercholesterolemic subjects. Funct Food Health Dis. 2011;1:106–17.
    1. Patel V, Rink C, Gordillo GM, et al. Oral tocotrienols are transported to human tissues and delay the progression of the model for end-stage liver disease score in patients. J Nutr. 2012;142:513–19.
    1. Ahmad NS, Khalid BA, Luke DA, Ima Nirwana S. Tocotrienol offers better protection than tocopherol from free radical-induced damage of rat bone. Clin Exp Pharmacol Physiol. 2005;32:761–70.
    1. Hermizi H, Faizah O, Ima-Nirwana S, et al. Beneficial effects of tocotrienol and tocopherol on bone histomorphometric parameters in sprague-dawley male rats after nicotine cessation. Calcif Tissue Int. 2009;84:65–74.
    1. Nasibah A, Rajikin MH, Nor-Ashikin MNK, Nuraliza AS. Tocotrienol improves the quality of impaired mouse embryos induced by corticosterone. Conf. Rec. Symposium on Humanities, Science and Engineering Research (SHUSER2012); 2012; pp. 135–38.
    1. Nasibah A, Rajikin MH, Nor-Ashikin MNK, Nuraliza AS. Effects of tocotrienol supplementation on pregnancy outcome in mice subjected to maternal corticosterone administration. Journal of Oil Palm Research. 2012;24:1550–58.
    1. Kamsani YS, Rajikin MH, Nor-Ashikin MNK, et al. Nicotine-induced cessation of embryonic development is reversed by γ-tocotrienol in mice. Med Sci Monit Basic Res. 2013;19:87–92.
    1. Mokhtar N, Rajikin MH, Zakaria Z. Role of tocotrienol-rich palm vitamin E on pregnancy and preimplantation embryos in nicotine treated rats. Biomed Res. 2008;19:181–84.
    1. Dobrinsky JR. Cellular approach to cryopreservation of embryos. Theriogenology. 1996;45:17–26.
    1. Feltes BC, de Faria Poloni J, Notari DL, Bonatto D. Toxicological effects of the different substances in tobacco smoke on human embryonic development by a systems chemo-biology approach. PLoS One. 2013;8:e61743.
    1. Khanna S, Patel V, Rink C, et al. Delivery of orally supplemented alpha-tocotrienol to vital organs of rats and tocopherol-transport protein deficient mice. Free Radic Biol Med. 2005;39:1310–19.
    1. Okabe M, Oji M, Ikeda I, et al. Tocotrienol levels in various tissues of sprague-dawley rats after intragastric administration of tocotrienols. Biosci Biotechnol Biochem. 2002;66:1768–71.
    1. Ikeda S, Tohyama T, Yoshimura H, et al. Dietary α-tocopherol decreases α-tocotrienol but not γ-tocotrienol concentration in rats. J Nutr. 2003;133:428–34.
    1. Suzuki YJ, Tsuchiya M, Wassall SR, et al. Structural and dynamic membrane properties of alpha-tocopherol and alpha-tocotrienol: Implication to the molecular mechanism of their antioxidant potency. Biochem. 1993;32:10692–99.
    1. Dasiman R, Rahman NS, Othman S, et al. Cytoskeletal alterations in different developmental stages of in vivo cryopreserved preimplantation murine embryos. Med Sci Monit Basic Res. 2013;19:258–66.
    1. Li L, Zheng P, Dean J. Maternal control of early mouse development. Development. 2010;137:859–70.
    1. Syairah SMM, Rajikin MH, Sharaniza AR, et al. Chromosomal status in murine preimplantation 2-cell embryos following annatto (Bixa orellana)-derived pure delta-tocotrienol supplementation in normal and nicotine-treated mice. World Appl Sci J. 2016;34:1855–59.

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