BODIPY-cholesterol can be reliably used to monitor cholesterol efflux from capacitating mammalian spermatozoa

N C Bernecic, M Zhang, B M Gadella, J F H M Brouwers, J W A Jansen, G J A Arkesteijn, S P de Graaf, T Leahy

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    Capacitation is the final maturation step spermatozoa undergo prior to fertilisation. The efflux of cholesterol from the sperm membrane to the extracellular environment is a crucial step during capacitation but current methods to quantify this process are suboptimal. In this study, we validate the use of a BODIPY-cholesterol assay to quantify cholesterol efflux from spermatozoa during in vitro capacitation, using the boar as a model species. The novel flow cytometric BODIPY-cholesterol assay was validated with endogenous cholesterol loss as measured by mass spectrometry and compared to filipin labelling. Following exposure to a range of conditions, the BODIPY-cholesterol assay was able to detect and quantify cholesterol efflux akin to that measured with mass spectrometry. The ability to counterstain for viability is a unique feature of this assay that allowed us to highlight the importance of isolating viable cells only for a reliable measure of cholesterol efflux. Finally, the BODIPY-cholesterol assay proved to be the superior method to quantify cholesterol efflux relative to filipin labelling, though filipin remains useful for assessing cholesterol redistribution. Taken together, the BODIPY-cholesterol assay is a simple, inexpensive and reliable flow cytometric method for the measurement of cholesterol efflux from spermatozoa during in vitro capacitation.

    Original languageEnglish
    Article number9804
    JournalScientific Reports
    Volume9
    Issue number1
    DOIs
    Publication statusPublished - 8 Jul 2019

    Fingerprint

    Dive into the research topics of 'BODIPY-cholesterol can be reliably used to monitor cholesterol efflux from capacitating mammalian spermatozoa'. Together they form a unique fingerprint.

    Cite this