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Dynamic Expression and New Functions of Early B Cell Factor 2 in Cerebellar Development

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Abstract

The collier/Olf1/EBF family genes encode helix-loop-helix transcription factors (TFs) highly conserved in evolution, initially characterized for their roles in the immune system and in various aspects of neural development. The Early B cell Factor 2 (Ebf2) gene plays an important role in the establishment of cerebellar cortical topography and in Purkinje cell (PC) subtype specification. In the adult cerebellum, Ebf2 is expressed in zebrin II (ZII)-negative PCs, where it suppresses the ZII+ molecular phenotype. However, it is not clear whether Ebf2 is restricted to a PC subset from the onset of its expression or is initially distributed in all PCs and silenced only later in the prospective ZII+ subtype. Moreover, the dynamic distribution and role of Ebf2 in the differentiation of other cerebellar cells remain unclarified. In this paper, by genetic fate mapping, we determine that Ebf2 mRNA is initially found in all PC progenitors, suggesting that unidentified upstream factors silence its expression before completion of embryogenesis. Moreover we show Ebf2 activation in an early born subset of granule cell (GC) precursors homing in the anterior lobe. Conversely, Ebf2 transcription is repressed in other cerebellar cortex interneurons. Last, we show that, although Ebf2 only labels the medial cerebellar nuclei (CN) in the adult cerebellum, the gene is expressed prenatally in projection neurons of all CN. Importantly, in Ebf2 nulls, fastigial nuclei are severely hypocellular, mirroring the defective development of anterior lobe PCs. Our findings further clarify the roles of this terminal selector gene in cerebellar development.

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Acknowledgments

Oocyte injections were performed at the Center for Conditional Mutagenesis (CFCM), San Raffaele Scientific Institute. Image analysis was carried out at ALEMBIC, an advanced microscopy laboratory established by the San Raffaele Scientific Institute and University.

Funding

G.G.C.’s research was funded by the Italian Telethon Foundation, grant GGP13146. O.C. was the recipient of a grant from the Italian Ministry of Health (Ministero della Salute Ricerca Finalizzata 2011-PE-2011-02347716). R.H. was supported by an award from the Canadian Institutes of Health Research.

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Authors and Affiliations

  1. Division of Neuroscience, San Raffaele Scientific Institute, Via Olgettina 60, Milan, 20132, Italy

    Aurora Badaloni, Filippo Casoni, Laura Croci, Francesca Chiara, Ottavio Cremona & G. Giacomo Consalez

  2. Università Vita-Salute San Raffaele, Via Olgettina 58, 20132, Milan, Italy

    Filippo Casoni, Ottavio Cremona & G. Giacomo Consalez

  3. Department of Basic Medical Sciences, Neurosciences and Sensory Organs, University of Bari Medical School, Piazza Giulio Cesare 11, Bari, 70124, Italy

    Antonella Bizzoca & Gianfranco Gennarini

  4. Department of Cell Biology and Anatomy and Hotchkiss Brain Institute, Cumming School of Medicine, University of Calgary, Calgary, Alberta, T2N 1N4, Canada

    Richard Hawkes

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  1. Aurora Badaloni
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  2. Filippo Casoni
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Correspondence to G. Giacomo Consalez.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The experimental plan was designed in agreement with the stipulations of the San Raffaele Institutional Animal Care and Use Committee (permit number 336).

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Badaloni, A., Casoni, F., Croci, L. et al. Dynamic Expression and New Functions of Early B Cell Factor 2 in Cerebellar Development. Cerebellum 18, 999–1010 (2019). https://doi.org/10.1007/s12311-019-01051-3

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