Characterization of Mitofusin-2 knock down in Sertoli cell line and its role in phagocytosis

Cereceda, K; Munoz, J. P.; Slebe J.C; Zorzano, A; Concha, I


Sertoli cells have multiple roles in germ cell development, ranging from physical support to supply of nutrients. The mechanisms that regulate Sertoli cell metabolism are central to the maintenance of spermatogenesis and male fertility. One of the functions of Sertoli cells corresponds to phagocytosis of residual bodies and apoptotic spermatogenic cells. It has been shown in phagocytes that there is a correlation between mitochondrial membrane potential and the engulfment capacity. Mitochondrial dynamics and quality control includes the exchange of mitochondrial components through mitochondrial fusion and fission. Mitofusin 2 (Mfn2) is a mitochondrial outer membrane protein involved in the rearrangement of these organelles through the regulation of the fusion process. In this work we evaluated the role of Mfn2 in the maintenance of mitochondrial and phagocytic function in Sertoli cells. We generated a Sertoli 42GPA9 Mitofusin 2 knockdown (KD)cell line and evaluated different parameters of mitochondrial function and observed an increase in ROS production, a decrease in mitochondrial membrane potential, higher calcium levels and a fragmented mitochondrial morphology, demonstrating an altered mitochondrial function. Phagocytosis was determined by incubating the cells with fluorescent beads and evaluated its internalization by microscopy and flow cytometry, which was reduced in a 30%. These results suggest that Mfn2 is involved in Sertoli cell phagocytosis

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Fecha de publicación: 2017
Año de Inicio/Término: 26-29 Septiembre, 2017