Characterization of Foxp2-expressing cells in the developing spinal cord

Two members of winged-helix/forkhead transcription factors, Foxp1 and Foxp2, are expressed in the developing and adult CNS, including the striatum, cerebral cortex, and thalamus. In a previous study, we have demonstrated that Foxp1 is expressed in a subpopulation of V1 interneurons in addition to motor neurons of the spinal cord during mouse embryogenesis. However, the detailed expression pattern of Foxp2 and its relationship with Foxp1 in the developing spinal cord remains to be elucidated. To shed light on the potential roles of Foxp1 and Foxp2 in the developing spinal cord, we characterized Foxp2-expressing cells during mouse embryogenesis. At embryonic day (E) 11.0, Foxp2-expressing cells were first observed in the ventral spinal cord, which were Pax6(-), p27(+), and neuron-specific class III beta-tubulin(+) postmitotic neurons. Between E13.5 and E15.5, high expression of Foxp2 was observed in both medial and lateral parts of the ventral spinal cord. Double-immunofluorescence staining for Foxp2 with some homeodomain transcription factors revealed that Foxp2-expressing neurons were Pax2(+), En1(+), Evx1(-), Chx10(-), Gata3(-), and Lhx3(-) V1 interneurons in the intermediate zone throughout the ventral spinal cord, indicating that Foxp2-expressing neurons were also V1 interneurons with the same phenotypes as Foxp1-expressing interneurons. In addition, neither Foxp1 nor Foxp2 was expressed in ventral calbindin(+) Renshaw cells. However, Foxp2 did not colocalize with Foxp1 in interneurons of the ventral spinal cord. These findings suggest that Foxp1 and Foxp2 are expressed in the distinct subsets of V1 interneurons that belong to non-Renshaw cells in the ventral spinal cord during embryogenesis. Thus, Foxp1 and Foxp2 may be involved in the determination of the cell type identities during late embryogenesis: the classes of neurotransmitters and the functional subtypes of non-Renshaw cells, such as Ia and Ib inhibitory interneurons.