🧫 Experiment Protocol
ValidationRab27aNeonatal mice with dorsal subventricular zone manipulationproposed
Neonatal electroporation and shRNA-mediated knockdown of Rab27a was performed in dorsal subventricular zone neural stem cells (NSCs) and astrocytes. This manipulation increased the number of CD11b/IBA1 positive rounded microglia, suggesting that disruption of EV release machinery affects microglial activation state. The experiment demonstrated that EV release pathways, particularly those involving Rab27a, are important for regulating microglial morphology and potentially activation in the developing nervous system.
PRIMARY OUTCOME
Changes in microglia morphology and number following Rab27a knockdown
EXPECTED OUTCOMES
Increased rounded microglia following disruption of EV release
SUCCESS CRITERIA
Significant increase in CD11b/IBA1 positive rounded microglia
PROTOCOL
Neonatal electroporation, shRNA-mediated knockdown, immunofluorescence analysis of microglia