Induction and Quantification of shRNA-Induced Cacna1g Knockdown in the Mesencephalic Locomotor Region of Mice

Locomotion is initiated in the brain through the activity of supraspinal networks that project to spinal cord central pattern generators. Of great importance in this pathway is the mesencephalic locomotor region (MLR), a command center that, when electrically stimulated in animal models, elicits and modulates locomotion. The molecular mechanisms, specifically the ion channels and ionic currents, that activate nuclei of the MLR and thus contribute to locomotor program initiation are incompletely understood. We hypothesize that glutamatergic neurons in the MLR become active following disinhibition, which allows MLR neurons to cause excitation of their postsynaptic targets and drive the execution of motor commands. To evaluate the role of excitation-after-inhibition dynamics in the MLR, we employ short hairpin RNA technology to conduct gene knockdown investigations and concurrent locomotor assessments in mice. Our work contributes to a more complete understanding of the midbrain circuits that gait locomotion.