Influence of ejaculate proteins from different canine breeds on quality and survival refrigerated sperm with or without seminal plasma

Abstract

The aim of the present work is to study canine breeds influence on protein profile of Seminal Plasma (SP) and sperm cell, as well as on cooled sperm survial and quality; besides evaluate SP influence on sperm quality and Reatives Oxygen Species (ROS) production during cooling semen procedure. Semen collection from four different canine breeds (n=20) will be performed at the first stage of the study for proteomic analysis (spermatozoa and SP) of each individual. In the second stage, pool of 5 animals of each the same breeds will be performed. Pools will be divided into 3 aliquots and the treatments will be composed of 1) semen centrifuged and without SP; 2) semen centrifuged and diluted in autologous SP; and 3) semen not centrifuged. The 3 samples will be evaluated, refrigerated with diluent medium and transported to the laboratory, where they will be kept at 5ºC for 168 hours. Complete semen analysis in all samples will be performed every 48 hours until 168 hours of refrigeration, as well as evaluation of spermatic membranes integrity by flow cytometry, mitochondrial potential and the production of mitochondrial superoxide. ROS production evaluation by TBARs (Thiobarbituric Acid) and antioxidant enzymes will also be performed. It is expected to observe an influence of the canine breeds on SP protein profile and sperm cells, with respect to semen survival during the refrigeration; and a change in ROS production due to the presence of SP during cooling semen. (AU)