Neurons that contact the cerebrospinal fluid (CSF-cNs) are a population of evolutionary conserved cells located around the central canal along the spinal cord. Their physiological role is poorly understood but recent works in lower vertebrates have proposed a regulatory function of spinal neuronal networks that define the locomotion. CSF-cNs have a short dendritic process that ends with a terminal bud in direct contact with the cerebrospinal fluid and express the PKD2L1 channel, a sensory transduction protein sensitive to variation of pH and osmolarity. Their unique anatomical structure, coupled with their sensory molecular apparatus, suggests that CSF-cNs adjust the locomotor activity after changes of cerebrospinal fluid composition or flow.
PKD2L1 are cationic channels that are able to provide sufficient depolarization to drive action potentials in CSF-cNs, which indicates that PKD2L1 channels have an important role in the control of CSF-cNs output. In this context, it is of interest to study the mechanisms underlying the regulation of PKD2L1 channels function and we propose that membrane interacting proteins may contribute to fine-tune the activity of the channel. By combining histolocalization, biochemical and electrophysiological approaches, the Master student will contribute to the identification of the molecular partners of PKD2L1 channels in spinal CSF-cNs of mouse and the study of their regulation of the channel activity as well as the CSF-cNs excitability.
• Patch-clamp recordings on acute spinal cord slices.
• Immunohistofluorescence and/or fluorescence in situ hybridization followed by observation with confocal imaging.
Co-immunoprecipitation from spinal cord lysates.