Enigmatic cerebrospinal fluid-contacting neurons arise even after the termination of neurogenesis in the rat spinal cord during embryonic development and retain their immature-like characteristics until adulthood.

Despite the abundance of cerebrospinal fluid-contacting neurons (CSF-cNs) lining the central canal of the spinal cord of mammals, little information is known regarding the phenotype and fate of these cells during development and in adulthood. Using immunofluorescence of spinal cord tissue of rats from the first postnatal day (P1) until the end of the 5th postnatal week (P36), we observed that these neurons show both immature (doublecortin+, β-III-tubulin+, neurofilament 200 kDa-) and more mature (weak NeuN+, P2X2+, GAD65+) characteristics during the first postnatal weeks. Because of the gradually decreasing number of CSF-cNs in the central canal lining during development, we were also interested in the migration potential of these cells. However, the assessment of the number of CSF-cNs in the lining of the central canal during postnatal development revealed that this decline is most likely associated with the growth of the spinal cord. Lastly, to reveal the birth date of CSF-cNs, we performed 5-bromo-2-deoxyuridine administration and colocalization analyses. We found that production of these cells appears from day 12 of embryonic development (E12) until E22. The vast majority of CSF-contacting neurons arise on E14 and E15. In contrast with other types of spinal neurons, the production of CSF-cNs is not restricted to a particular neuroepithelial region and occurs even after what is thought to be the termination of neurogenesis.