Efficiency of CellROX deep red (R) and CellROX orange (R) fluorescent probes in identifying reactive oxygen species in sperm samples from high and low fertility bulls

Author(s)

Carvalho Celeghini, Eneiva Carla ^[1] ; Rodrigues Alves, Maira Bianchi ^[1] ; de Arruda, Rubens Paes ^[1] ; de Rezende, Gabriela Marques ^[1] ; Florez-Rodriguez, Shirley Andrea ^[1] ; de Sa Filho, Manoel Francisco ^{[1, 2]}

Affiliation

^[1] Univ Sao Paulo, Sch Vet Med & Anim Sci, Dept Anim Reprod, St Duque Caxias Norte 225, BR-13635900 Pirassununga - Brazil

^[2] Alta Genet, Uberaba - Brazil

• Document type: Journal article
• Source: ANIMAL BIOTECHNOLOGY; v. 32, n. 1 AUG 2019.
• Web of Science Citations: 10

Abstract

Detection of reactive oxygen species (ROS) is of great interest in semen analysis since their excess is detrimental to sperm function and male fertility. Fluorescence microscopy has achieved attention for providing broad possibilities of sperm evaluations and also for presenting substantial accessibility. In this context, this study investigated the efficiency of CellROX Deep Red (R) and Orange (R) probes in detecting ROS in bovine sperm cells and assessed their relationship with sperm fertility potential. First, 16 ejaculates were assigned in three treatments: T0 (no ROS production induced), T1x (ROS production induced once) and T2x (ROS production induced twice). Samples were incubated with Red and Orange probes and percentages of cells producing ROS were evaluated using fluorescence microscopy. Coefficient of determination was 0.61 for Red and 0.56 for Orange. Afterwards, frozen-thawed semen samples from high and low fertility bulls were evaluated regarding percentages of cells producing ROS detected by Red and Orange. Higher levels of ROS assessed by Red were detected in low fertility bovine samples. In conclusion, CellROX Red (R) and Orange (R) are both efficient in detecting ROS in bovine spermatozoa. Furthermore, higher sperm ROS detection by CellROX Red (R) might be associated with low fertility samples. (AU)

• FAPESP's process: 15/09154-6 - Profile of microRNAs present in semen and embryos and their relationship to fertility
• Grantee: Maíra Bianchi Rodrigues Alves
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/05395-1 - Bull semen and embryos microRNAs profile and these correlation with fertility
• Grantee: Eneiva Carla Carvalho Celeghini
• Support Opportunities: Regular Research Grants