HPA Stress Axis Responsivity Assessment
Objective: Measurement of offspring hypothalamic-pituitary-adrenal (HPA) stress axis responsivity to stress exposure following paternal stress exposure
Protocol Steps
Paternal Stress Exposure
Male mice were exposed to chronic stress before breeding. Stress exposure occurred in two groups: either throughout puberty or in adulthood.
Note: Two separate paternal stress exposure groups were used to examine variation in germ cell susceptibility across the lifespan
View evidence from paper
“we exposed mice to 6 weeks of chronic stress before breeding. male stress exposure occurred either throughout puberty or in adulthood”
Breeding
Stressed males were bred to produce offspring for HPA axis assessment
Note: Offspring from both paternal stress groups were examined
View evidence from paper
“offspring of sires from both paternal stress groups displayed significantly reduced HPA stress axis responsivity”
HPA Stress Axis Responsivity Assessment
Offspring HPA stress axis responsivity was measured in response to stress exposure
Note: Specific methodology for stress exposure and measurement not detailed in provided text
View evidence from paper
“offspring of sires from both paternal stress groups displayed significantly reduced HPA stress axis responsivity”
Brain Tissue Collection and Gene Expression Analysis
Gene set enrichment analyses were performed in offspring stress-regulating brain regions including the paraventricular nucleus (PVN) and bed nucleus of stria terminalis
Note: Analysis revealed global pattern changes in transcription suggestive of epigenetic reprogramming
View evidence from paper
“Gene set enrichment analyses in offspring stress regulating brain regions, the paraventricular nucleus (PVN) and the bed nucleus of stria terminalis, revealed global pattern changes in transcription”
Sperm MicroRNA Analysis
Sperm microRNA content was examined in stressed males to identify potential epigenetic mechanisms of germ cell transmission
Note: Nine specific microRNAs were significantly increased in both paternal stress groups
View evidence from paper
“we found robust changes in sperm microRNA (miR) content, where nine specific miRs were significantly increased in both paternal stress groups”