Odor Fear Conditioning
Objective: To examine the inheritance of parental traumatic exposure by subjecting F0 mice to odor fear conditioning before conception and measuring behavioral sensitivity and neuroanatomical changes in F1 and F2 generations
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Materials1
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Protocol Steps
F0 Odor Fear Conditioning
F0 mice were subjected to odor fear conditioning before conception using acetophenone as the conditioned odor stimulus
Note: Conditioning performed on F0 generation prior to mating
View evidence from paper
“We subjected F0 mice to odor fear conditioning before conception”
F1 and F2 Generation Behavioral Testing
Subsequently conceived F1 and F2 generations were tested for behavioral sensitivity to the F0-conditioned odor (acetophenone) and other odors
Note: Behavioral sensitivity measured in naive offspring without prior conditioning
View evidence from paper
“subsequently conceived F1 and F2 generations had an increased behavioral sensitivity to the F0-conditioned odor, but not to other odors”
Neuroanatomical Analysis
Enhanced neuroanatomical representation of the Olfr151 pathway was examined in F1 and F2 generations
Note: Neuroanatomical changes complemented behavioral sensitivity findings
View evidence from paper
“the behavioral sensitivity of the F1 and F2 generations to acetophenone was complemented by an enhanced neuroanatomical representation of the Olfr151 pathway”
Sperm DNA Bisulfite Sequencing
Bisulfite sequencing was performed on sperm DNA from conditioned F0 males and F1 naive offspring to examine CpG methylation patterns in the Olfr151 gene
Note: Analysis revealed CpG hypomethylation in the Olfr151 gene
View evidence from paper
“Bisulfite sequencing of sperm DNA from conditioned F0 males and F1 naive offspring revealed CpG hypomethylation in the Olfr151 gene”
In Vitro Fertilization and Cross-Fostering Studies
In vitro fertilization, F2 inheritance studies, and cross-fostering experiments were conducted to determine the route of transgenerational inheritance
Note: These experiments demonstrated that transgenerational effects are inherited via parental gametes
View evidence from paper
“In addition, in vitro fertilization, F2 inheritance and cross-fostering revealed that these transgenerational effects are inherited via parental gametes”