Evaluating Mitochondrial Membrane Potential in Cryopreserved Semen of Toraya Buffalo through Flow Cytometry

This paper was not presented at the conference.

Abstract

This paper was not presented at the conference.

Cryopreservation significantly reduces sperm quality, including mitochondrial membrane potential (MMP), in buffalo bull semen. This study aimed to examine mitochondrial membrane potential using the fluorescent dyes JC-1 and Mitostatus in cryopreserved semen of Toraya buffalo, employing the flow cytometry method. Frozen semen was obtained from seven Toraya buffalo bulls aged 4-8 years. Quantitative assessment of buffalo sperm MMPs was conducted using the fluorescent dyes JC-1 (5,5,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazoylcarbocyanine iodide) and MitoStatus TMRE (Tetramethylrhodamine ethyl ester). Flow cytometry analysis was performed using a BD Accuri 6 plus with a 488 nm laser. Data were analyzed using an independent T-test in Minitab 18.0. The results showed no difference in MMP evaluation using JC-1 and Mitostatus (P<0.05). In conclusion, The combined use of JC-1 staining and flow cytometry has been recognized as an effective approach for assessing mitochondrial membrane potential (MMP) in buffalo sperm. Additionally, Mitostatus TMRE has been identified as a viable alternative method for evaluating MMP in buffalo sperm