Cerebellar Craniotomy for In Vivo Calcium Imaging of Astrocytes
Adapted from Imaging in Neuroscience (ed. Helmchen and Konnerth). CSHL Press, Cold Spring Harbor, NY, USA, 2011.Abstract
The cerebellar cortex contains two astrocyte types: the Bergmann glia of the molecular layer and the velate protoplasmic astrocytes of the granule cell layer. In vivo, these cell types generate both subcellular calcium transients and trans-glial calcium waves. It is possible to perform in vivo calcium imaging in cerebellar astrocytes. One method involves injection of a replication-incompetent recombinant adenovirus for gene transfer of a fluorescent calcium indicator protein. A second method uses multicell bolus loading (MCBL) in the molecular layer of the cerebellum with synthetic calcium indicators. This protocol presents a cerebellar craniotomy procedure which can be used to prepare a virus-injected animal for in vivo imaging. It can also be used to prepare an animal for MCBL.
- © 2011 Cold Spring Harbor Laboratory Press
