Effect of LPS on In Vitro Embryo Development in Bubalus bubalis: A Model for Studying Inflammatory Pathways.

Area of Science:

• Veterinary Reproduction and Embryology
• Molecular Biology
• Animal Science

Background:

• Reproductive efficiency in buffalo is vital for sustainable agriculture.
• Endotoxin lipopolysaccharide (LPS) triggers inflammatory responses affecting female fertility and embryo development.
• Endometritis serves as an in vitro model to study LPS effects on embryo competence.

Purpose of the Study:

• To investigate the molecular mechanisms by which LPS impacts buffalo embryo competence.
• To evaluate the effects of LPS on oocyte and embryo development in vitro.
• To elucidate the role of inflammatory and oxidative stress pathways in LPS-induced developmental defects.

Main Methods:

• Buffalo ovaries were sourced, and oocytes were aspirated.
• Oocytes underwent in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC).
• Media were supplemented with 5 μg/mL lipopolysaccharide (LPS) during IVC.

Main Results:

• LPS exposure significantly reduced cleavage rates, blastocyst formation, and total cell count.
• Mitochondrial membrane potential (MMP) and trophectoderm: inner cell mass (TE:ICM) ratio decreased, while reactive oxygen species (ROS) and apoptosis increased.
• LPS upregulated inflammatory (TLR4, CD14, RPS27A) and apoptotic (Caspase 3) genes, and downregulated antioxidant (GPX1) and pluripotent (Oct4) genes.

Conclusions:

• Lipopolysaccharide (LPS) at 5 μg/mL adversely affects buffalo embryonic development during in vitro culture (IVC).
• LPS induces oxidative stress, apoptosis, and alters key gene expression, negatively impacting embryo quality.
• These findings highlight the detrimental effects of inflammatory stimuli on buffalo reproductive outcomes.