Purkinje cells originate from cerebellar ventricular zone progenitors positive for Neph3 and E-cadherin

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ydbio.2009.11.032 Byline: Eri Mizuhara (a), Yasuko Minaki (a), Tomoya Nakatani (a), Minoru Kumai (b), Takeshi Inoue (a), Keiko Muguruma (c), Yoshiki Sasai (c), Yuichi Ono (a)(b) Keywords: Neph3; E-cadherin; Ptf1a; Purkinje cell; Origin; Neuronal subtype; Cell sorting; Cell surface marker; Fate mapping Abstract: GABAergic Purkinje cells (PCs) provide the primary output from the cerebellar cortex, which controls movement and posture. Although the mechanisms of PC differentiation have been well studied, the precise origin and initial specification mechanism of PCs remain to be clarified. Here, we identified a cerebellar and spinal cord GABAergic progenitor-selective cell surface marker, Neph3, which is a direct downstream target gene of Ptf1a, an essential regulator of GABAergic neuron development. Using FACS, Neph3.sup.+ GABAergic progenitors were sorted from the embryonic cerebellum, and the cell fate of this population was mapped by culturing in vitro. We found that most of the Neph3.sup.+ populations sorted from the mouse E12.5 cerebellum were fated to differentiate into PCs while the remaining small fraction of Neph3.sup.+ cells were progenitors for Pax2.sup.+ interneurons, which are likely to be deep cerebellar nuclei GABAergic neurons. These results were confirmed by short-term in vivo lineage-tracing experiments using transgenic mice expressing Neph3 promoter-driven GFP. In addition, we identified E-cadherin as a marker selectively expressed by a dorsally localized subset of cerebellar Neph3.sup.+ cells. Sorting experiments revealed that the Neph3.sup.+ E-cadherin.sup.high population in the embryonic cerebellum defined PC progenitors while progenitors for Pax2.sup.+ interneurons were enriched in the Neph3.sup.+ E-cadherin.sup.low population. Taken together, our results identify two spatially demarcated subregions that generate distinct cerebellar GABAergic subtypes and reveal the origin of PCs in the ventricular zone of the cerebellar primordium. Author Affiliation: (a) Group for Neuronal Differentiation, KAN Research Institute Inc., Kobe, Hyogo 650-0047, Japan (b) Group for Transgenic Technology, KAN Research Institute Inc., Kobe, Hyogo 650-0047, Japan (c) Organogenesis and Neurogenesis Group, Center for Developmental Biology, RIKEN, Kobe, Hyogo 650-0047, Japan Article History: Received 30 July 2009; Revised 12 November 2009; Accepted 30 November 2009