Oxidative Stress Dysregulates Protein Homeostasis Within the Male Germ Line

Oxidative stress is causally linked to male reproductive pathologies, driven primarily by lipid peroxidation and an attendant production of highly reactive lipid aldehydes, such as 4-hydroxynonenal (4HNE) within the male germ line. In somatic cells, 4HNE dysregulates proteostasis targeting of vulnerable proteins for adduction, causing protein misfolding and eventually aggregation. The aims of this study were to explore whether oxidative stress precipitates an equivalent response in the male germ line and determine the protective mechanisms used by germ cells to prevent this cascade of protein damage. We reveal a causative role for oxidative stress in the accumulation of protein deposits in male germ cells. Specifically, 4HNE treatment resulted in a significant increase in cytosolic protein aggregation within pre- and post-meiotic germ cells as measured by the aggregate-detecting fluorophores ProteoStat and Thioflavin T, and the amyloid-specific anti-A11 and anti-OC antibodies. Our data implicate nucleocytoplasmic transport machinery and molecular chaperones as potential mechanisms for the subcellular compartmentalization and/or suppression of aggregating proteins. Thus, the inhibition of karyopherin transport proteins and molecular chaperones resulted in a significant increase in the accumulation of aggregated cellular protein. These data establish the novel paradigm that lipid peroxidation is a key contributor to a decline in proteostasis in developing germ cells. These findings will inform the development of novel strategies to protect germ cells from oxidative stress. Together, these results shed light on proteostasis mechanisms that may assist in the management of misfolded proteins in the male germ line under conditions of acute oxidative stress.