AUTHOR=Turner Katerina A. , Fishman Emily L. , Asadullah Mariam , Ott Brooke , Dusza Patrick , Shah Tariq A. , Sindhwani Puneet , Nadiminty Nagalakshmi , Molinari Emanuela , Patrizio Pasquale , Saltzman Barbara S. , Avidor-Reiss Tomer 

TITLE=Fluorescence-Based Ratiometric Analysis of Sperm Centrioles (FRAC) Finds Patient Age and Sperm Morphology Are Associated With Centriole Quality

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=9

YEAR=2021

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2021.658891

DOI=10.3389/fcell.2021.658891

ISSN=2296-634X

ABSTRACT=<p>A large proportion of infertility and miscarriage causes are unknown. One potential cause is a defective sperm centriole, a subcellular structure essential for sperm motility and embryonic development. Yet, the extent to which centriolar maladies contribute to male infertility is unknown due to the lack of a convenient way to assess centriole quality. We developed a robust, location-based, ratiometric assay to overcome this roadblock, the Fluorescence-based Ratiometric Assessment of Centrioles (FRAC). We performed a case series study with semen samples from 33 patients, separated using differential gradient centrifugation into higher-grade (pellet) and lower-grade (interface) sperm fractions. Using a reference population of higher-grade sperm from infertile men with morphologically standard sperm, we found that 79% of higher-grade sperm of infertile men with substandard sperm morphology have suboptimal centrioles (<italic>P</italic> = 0.0005). Moreover, tubulin labeling of the sperm distal centriole correlates negatively with age (<italic>P</italic> = 0.004, <italic>R</italic> = −0.66). These findings suggest that FRAC is a sensitive method and that patient age and sperm morphology are associated with centriole quality.</p>