Changes in expression and distribution of the glutamate transporter EAAT4 in developing mouse Purkinje cells

EAAT4 is a Purkinje cell (PC)-specific, postsynaptic glutamate transporter in the adult mouse brain. Here, we performed developmental analyses to reveal its temporal expression in relation to PC differentiation. Using in situ hybridization, EAAT4 mRNA was specifically detected in the PC layer of the cerebellar primordium at embryonic day 13 (E13). During late fetal and neonatal periods, the transcripts were detected only in the PC layer in the caudal cerebellum. At postnatal day 7 (P7) and, thereafter, the prominent transcripts were found on monolayered PCs in the entire cerebellum. Using immunohistochemistry, low levels of EAAT4 immunoreactivity were first observed at E18 in the perikarya of PCs in the caudal cerebellum, and this pattern of immunostaining was maintained at P1 and P7. At P14 and, thereafter, the molecular layer in the entire cerebellum became immunopositive for EAAT4, and the intense immunoreactivity was detected preferentially in PC spines synapsing with parallel fiber terminals. Therefore, the present study has clarified that the transcription of EAAT4 begins in PCs from early embryonic stages, and that the synaptic localization of EAAT4 is established during the second postnatal week. When considered in the light of the synaptogenesis of parallel fiber-PC synapses which actively occurs in the rodent cerebellum during the second and third weeks of life, synaptic localization of the glutamate transporter EAAT4 may be closely associated with the synapse formation.