Effect of Melatonin-loaded Chitosan Nanoparticles (CMN) on Gene Expression of In-vitro Matured Buffalo Oocyte

Abstract

This study aimed to evaluate the effect of melatonin and melatonin-loaded chitosan nanoparticle (CMN) supplementation to maturation media on buffalo oocyte maturation rate and relative expression of genes: growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), B Cell Lymphoma 2 (BCL2), Associated X protein (BAX) and superoxide dismutase 1 (SOD1). Buffalo ovaries were heaved from Al-Mounib slaughterhouse, cumulus-oocyte complexes (COCs) were in vitro matured in three different media, TCM-199 medium (control), TCM-199 with melatonin 10-9M, and TCM-199 with CMN 10-9M. The assessment of the nuclear maturation rate was carried out through the presence of the first polar body. In addition, the mature buffalo oocytes were stored on RNA later for genetic analysis of GDF9, BMP15, SOD1, BCL2, and BAX genes using quantitative real-time PCR (qRT-PCR). The results were reported that buffalo oocytes supplemented with melatonin-loaded chitosan nanoparticle (CMN) or melatonin have a significant effect on nuclear maturation rate 94.04±0.65 and 88.74±0.77 respectively when compared with buffalo oocytes matured with basic media (control) 79.67±1.35. Furthermore, buffalo oocytes supplemented with melatonin-loaded chitosan nanoparticle (CMN) or melatonin showed significant upregulation of GDF9, BMP15, SOD1, and BCL2 genes and significant downregulation of BAX gene when compared with oocyte matured with basic media (control). In conclusion, the results of nuclear maturation rate and relative expression pattern of GDF9, BMP15, SOD1, BCL2, and BAX reflect that melatonin-loaded chitosan nanoparticle (CMN) and melatonin` may play an important role in the buffalo oocytes developmental competence.

Keywords

• Buffalo oocyte
• Melatonin-loaded chitosan nanoparticle (CMN)
• In-vitro maturation
• Relative gene expression
• Melatonin