Developmental molecular signatures define de novo cortico-brainstem circuit for skilled forelimb movement.

Skilled movement relies on descending cortical projections to the brainstem and spinal cord. While corticospinal neurons (CSN) have long been recognized for their role in fine motor control, the contribution of cortical projections to the brainstem remains poorly understood. Here, we identify a previously unrecognized direct cortico-brainstem circuit that emerges early in development and persists into adulthood. A subset of subcerebral projection neurons (SCPN) limit their projections to the brainstem from the earliest stages of axon extension without ever extending to the spinal cord. Using FACS purification and single-cell RNA sequencing, we show that these cortico-brainstem neurons (CBN) can be prospectively identified by the expression of Neuropeptide Y (Npy) in development. Functional silencing of Npy+ CBN in adulthood leads to impaired skilled forelimb reaching, demonstrating their essential role in adult motor control. Npy+ CBN project preferentially to rostral brainstem regions, including the midbrain reticular formation. These findings reveal developmental molecular signatures that define cortico-brainstem pathways for adult skilled movement. Our work provides new insights into the developmental logic that establishes descending cortical circuits and opens avenues for targeted investigation of their roles in motor function and recovery after injury.