A Four-Group Comparative Chemogenetic Protocol for Dissecting Neural Circuit Contributions to Non-Empirical Representational Consolidation: Habit Rigidity, Agency Attribution, Belief Updating, and Thalamic Filtering in Mus musculus

Non-empirical representational consolidation(NERC) is the synaptic encoding of propositionalcontent driven by endogenously generated saliencesignals in the absence of correlated externalevidence. Two mechanistically distinct hypotheseshave been advanced regarding the primary neuralgenerators of NERC: the input suppressionhypothesis (PPC + NAcc + ACC) and the flexibilitydeficit hypothesis (lOFC + PrL/IL + MD + DLS). This paper specifies a five-group comparativeDREADD protocol in C57BL/6J mice (N=50, n=10/group)testing both hypotheses simultaneously: Group 1: PrL inhibition + lOFC/MD activation + DLS inhibitionGroup 2: PrL activation + lOFC/MD activation + DLS inhibitionGroup 3: PPC inhibition + NAcc inhibition + ACC activationGroup 4: PPC inhibition onlyGroup 5: Empty vector control The protocol resolves the PrL directionalityquestion, identifies the minimum effectivecircuit intervention, and specifies thetranslational path to prime editing viaAAV-PHP.eB and MRgFUS delivery. Vector design: AAV-hSyn/CaMKII-DREADD-T2A-fluorophore with WPRE. Distinct serotypesper injection site to prevent immune competition.Ligands: CNO (1.0 mg/kg) and DCZ (0.1 mg/kg). Companion preprints:Version 1 (theoretical framework):doi: 10.5281/zenodo.20236971Version 2 (quantitative threshold protocol):[doi version 2]