Functional regionalization of the differentiating cerebellar Purkinje cell population occurs in an activity-dependent manner

Alessandro Dorigo; Komali Valishetti; Florian Hetsch; Hideaki Matsui; Jochen C Meier; Kazuhiko Namikawa; Reinhard W Köster

doi:10.3389/fnmol.2023.1166900

Functional regionalization of the differentiating cerebellar Purkinje cell population occurs in an activity-dependent manner

Front Mol Neurosci. 2023 Apr 27:16:1166900. doi: 10.3389/fnmol.2023.1166900. eCollection 2023.

Authors

Alessandro Dorigo¹, Komali Valishetti¹, Florian Hetsch^{2

3}, Hideaki Matsui^{1

4}, Jochen C Meier², Kazuhiko Namikawa¹, Reinhard W Köster¹

Affiliations

¹ Cellular and Molecular Neurobiology, Technische Universität Braunschweig, Braunschweig, Germany.
² Cell Physiology, Zoological Institute, Technische Universität Braunschweig, Braunschweig, Germany.
³ Institute of Pathophysiology, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
⁴ Department of Neuroscience of Disease, Brain Research Institute, Niigata University, Niigata, Japan.

Abstract

Introduction: The cerebellum is organized into functional regions each dedicated to process different motor or sensory inputs for controlling different locomotor behaviors. This functional regionalization is prominent in the evolutionary conserved single-cell layered Purkinje cell (PC) population. Fragmented gene expression domains suggest a genetic organization of PC layer regionalization during cerebellum development. However, the establishment of such functionally specific domains during PC differentiation remained elusive.

Methods and results: We show the progressive emergence of functional regionalization of PCs from broad responses to spatially restricted regions in zebrafish by means of in vivo Ca2+-imaging during stereotypic locomotive behavior. Moreover, we reveal that formation of new dendritic spines during cerebellar development using in vivo imaging parallels the time course of functional domain development. Pharmacological as well as cell-type specific optogenetic inhibition of PC neuronal activity results in reduced PC dendritic spine density and an altered stagnant pattern of functional domain formation in the PC layer.

Discussion: Hence, our study suggests that functional regionalization of the PC layer is driven by physiological activity of maturing PCs themselves.

Keywords: Purkinje cell; calcium imaging; cerebellum; functional regionalization; optogenetics; spine density; zebrafish.

Grants and funding

The authors gratefully acknowledge funding of this project by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation–Projektnummer 241961032 to RK), the Volkswagenstiftung (project HOMEO-HIRN, ZN3673, to RK and JM), funds of Lower Saxony, Era Net Neuron II project CIPRESS (to JM), and an Alexander von Humboldt Fellowship (to HM).