Two photon imaging of calcium responses in murine Purkinje neurons

Sreeja Kumari Dhanya; Gaiti Hasan

doi:10.1016/j.xpro.2021.101105

Two photon imaging of calcium responses in murine Purkinje neurons

STAR Protoc. 2022 Jan 20;3(1):101105. doi: 10.1016/j.xpro.2021.101105. eCollection 2022 Mar 18.

Authors

Sreeja Kumari Dhanya^{1

2}, Gaiti Hasan¹

Affiliations

¹ National Centre for Biological Science, Tata Institute of Fundamental Research, Bangalore 560065, India.
² Sastra University, Thirumalaisamudram, Thanjavur, Tamil Nadu 613401, India.

Abstract

Purkinje neurons (PNs) are an important component of the motor learning and coordination circuit and are affected in spino-cerebellar ataxias. Maintaining healthy PNs in cerebellar slices and recording their Ca²⁺ transients can be challenging. Here, we describe a protocol for measuring Ca²⁺ transients in PNs from adult mice, including problem-solving tips. This protocol can be used to measure neuronal excitability and agonist-mediated Ca²⁺ signaling in cerebellar slices expressing a genetic Ca²⁺ reporter in all PNs, thus improving yield of data. For complete details on the use and execution of this profile, please refer to Dhanya and Hasan (2021).

Keywords: Cell Biology; Genetics; Microscopy; Molecular Biology; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Calcium / metabolism*
Calcium Signaling
Mice
Neurons / metabolism*
Photons
Purkinje Cells / metabolism*

Substances

Calcium