HPA Axis Stress Response Assessment
Objective: To assess whether central methionine infusion reverses the effects of maternal programming on HPA axis stress responses in adult offspring by measuring DNA methylation, GR expression, and behavioral/physiological stress responses
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Equipment1
Not specified • Not specified • Not specified • Not mentioned
Materials1
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Protocol Steps
Central methionine infusion
Adult offspring received central infusion of L-methionine, a precursor to S-adenosyl-methionine that serves as methyl group donor for DNA methylation
Note: Infusion was performed in adult animals to test reversibility of early-life epigenetic programming
View evidence from paper
“we centrally infused the adult offspring with the essential amino acid l-methionine, a precursor to S-adenosyl-methionine”
Stress response assessment
Measurement of hypothalamic-pituitary-adrenal axis responses to stress following methionine treatment
Note: Assessment included both physiological and behavioral stress responses
View evidence from paper
“methionine infusion reverses the effect of maternal behavior on DNA methylation, nerve growth factor-inducible protein-A binding to the exon 1 7 promoter, GR expression, and hypothalamic-pituitary-adrenal and behavioral responses to stress”
DNA methylation analysis
Examination of DNA methylation patterns at the hippocampal exon 1 7 glucocorticoid receptor promoter
Note: Assessed whether methionine infusion reversed maternal programming effects on epigenomic marking
View evidence from paper
“methionine infusion reverses the effect of maternal behavior on DNA methylation”
Glucocorticoid receptor expression measurement
Quantification of GR expression levels in hippocampus following methionine treatment
Note: GR expression was assessed as indicator of epigenetic reversal
View evidence from paper
“methionine infusion reverses the effect of maternal behavior on DNA methylation, nerve growth factor-inducible protein-A binding to the exon 1 7 promoter, GR expression”
Nerve growth factor-inducible protein-A binding assessment
Measurement of nerve growth factor-inducible protein-A binding to the exon 1 7 glucocorticoid receptor promoter
Note: Used as indicator of epigenomic state changes
View evidence from paper
“methionine infusion reverses the effect of maternal behavior on DNA methylation, nerve growth factor-inducible protein-A binding to the exon 1 7 promoter”