Exploratory experiment designed to discover new patterns targeting PACAP in mice with viral-mediated PACAP knockdown in mPFC. Primary outcome: ZG therapeutic efficacy and synaptic protein expression following PACAP knockdown
Viral-mediated PACAP knockdown vectors were transfected into the medial prefrontal cortex to specifically reduce PACAP expression and test its necessity for ZG's therapeutic effects. Following PACAP knockdown, mice were treated with ZG and assessed for behavioral improvements and synaptic protein expression changes. This loss-of-function experiment was crucial to demonstrate that PACAP signaling is required for ZG's beneficial effects on both insomnia-anxiety behaviors and synaptic plasticity.
Viral vector transfection into mPFC, PACAP knockdown confirmation, ZG treatment, behavioral and molecular assessments
PACAP knockdown would abolish ZG's therapeutic effects and prevent synaptic protein upregulation
Loss of ZG efficacy and reduced synaptic protein expression in PACAP knockdown animals
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