Organelle-specific probe JC-1 identifies membrane potential differences in the mitochondrial function of bovine sperm

Autor: D. L. Garner, C.A. Thomas
Rok vydání: 1999
Předmět:
Zdroj: Molecular Reproduction and Development. 53:222-229
ISSN: 1098-2795
1040-452X
DOI: 10.1002/(sici)1098-2795(199906)53:2<222::aid-mrd11>3.0.co;2-l
Popis: The mitochondrial probe 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyan ine iodide (JC-1) not only identifies mitochondria exhibiting low membrane potentials by the emission of green fluorescence (range, 510-520 nm) but also differentiates these from mitochondria exhibiting relatively high membrane potentials. This discrimination occurs because JC-1 forms aggregates at high membrane potentials. These J-aggregates emit a bright red-orange fluorescence at 590 nm. In this study, JC-1 was combined with the classical dead cell stain, propidium iodide (PI), to identify a spectrum of functional sperm along with degenerate sperm. Flow cytometric analysis of bull sperm showed that the aggregate:monomer ratio differed among bulls before cryopreservation (P 0.05). The effects of stain equilibration time, sperm concentration, and live:dead ratios were examined. The addition of SYBR-14 to the JC-1 and PI combination enhanced the distinction between the red PI-stained and red-orange JC-1-stained populations. This discrimination between J-aggregates and the PI-stained sperm was affected by sperm concentration. These studies show that JC-1 can be useful in monitoring mitochondrial function in bovine sperm.
Databáze: OpenAIRE
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