Alterations in sperm RNAs persist after alcohol cessation and correlate with epididymal mitochondrial dysfunction

Abstract

BackgroundChronic preconception paternal alcohol use adversely modifies the sperm epigenome, inducing fetoplacental and craniofacial growth defects in the offspring of exposed males. A crucial outstanding question in the field of paternal epigenetic inheritance concerns the resilience of the male germline and its capacity to recover and correct sperm‐inherited epigenetic errors after stressor withdrawal.ObjectivesWe set out to determine if measures of the sperm‐inherited epigenetic program revert to match the control treatment 1 month after withdrawing the daily alcohol treatments.Materials and methodsUsing a voluntary access model, we exposed C57BL/6J males to 6% or 10% alcohol for 10 weeks, withdrew the alcohol treatments for 4 weeks, and used RNA sequencing to examine gene expression patterns in the caput section of the epididymis. We then compared the abundance of sperm small RNA species between treatments.ResultsIn the caput section of the epididymis, chronic alcohol exposure induced changes in the transcriptional control of genetic pathways related to the mitochondrial function, oxidative phosphorylation, and the generalized stress response (EIF2 signaling). Subsequent analysis identified region‐specific, alcohol‐induced changes in mitochondrial DNA copy number across the epididymis, which correlated with increases in the mitochondrial DNA content of alcohol‐exposed sperm. Notably, in the corpus section of the epididymis, increases in mitochondrial DNA copy number persisted 1 month after alcohol cessation. Analysis of sperm noncoding RNAs between control and alcohol‐exposed males 1 month after alcohol withdrawal revealed a ∼100‐fold increase in mir‐196a, a microRNA induced as part of the nuclear factor erythroid 2‐related factor 2 (Nrf2)‐driven cellular antioxidant response.Discussion and conclusionOur data reveal that alcohol‐induced epididymal mitochondrial dysfunction and differences in sperm noncoding RNA content persist after alcohol withdrawal. Further, differences in mir‐196a and sperm mitochondrial DNA copy number may serve as viable biomarkers of adverse alterations in the sperm‐inherited epigenetic program.

Chronic paternal alcohol exposures induce dose-dependent changes in offspring craniofacial shape and symmetry

“…We postulate that sperm-inherited epigenetic factors alter the embryonic control of mitochondrial and oxidative stress-related genetic pathways, disrupting neural crest cell function. Enrichment of these same mitochondrial and redox-related pathways in the reproductive tract and sperm noncoding RNA profile of alcohol-exposed males support this assertion ( Roach et al, 2023 ).…”