Protocol for measurement of calcium dysregulation in human induced pluripotent stem cell-derived dopaminergic neurons

Xiling Yin; Jungwoo Wren Kim; Shiyu Liu; Ted M Dawson; Valina L Dawson

doi:10.1016/j.xpro.2021.100405

Protocol for measurement of calcium dysregulation in human induced pluripotent stem cell-derived dopaminergic neurons

STAR Protoc. 2021 Mar 23;2(2):100405. doi: 10.1016/j.xpro.2021.100405. eCollection 2021 Jun 18.

Authors

Xiling Yin^{1

2}, Jungwoo Wren Kim^{1

3

4}, Shiyu Liu¹, Ted M Dawson^{1

2

5

6

7

8}, Valina L Dawson^{1

2

3

5

7

8}

Affiliations

¹ Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
² Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
³ Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁴ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
⁵ Soloman H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁶ Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁷ Adrienne Helis Malvin Medical Research Foundation, New Orleans, LA 70130, USA.
⁸ Diana Helis Henry Medical Research Foundation, New Orleans, LA 70130, USA.

Abstract

Calcium regulation is a critical process in neurons, and Ca²⁺ signaling is a major contributor to neurological disorders including Parkinson's disease (PD). Here, combining calcium imaging with whole-cell Ca²⁺ current recording, we provide a detailed protocol for measuring Ca²⁺ homeostasis in dopaminergic (DA) neurons derived from human induced pluripotent stem cells (hiPSCs). This approach can be applied to investigate the role of Ca²⁺ homeostasis in neuronal functionality as well as in disease processes. For complete details on the use and execution of this protocol, please refer to Kim et al. (2020).

Keywords: Cell Differentiation; Microscopy; Neuroscience; Stem Cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Calcium / metabolism*
Calcium Signaling / physiology*
Dopaminergic Neurons* / cytology
Dopaminergic Neurons* / pathology
Electrophysiology / methods*
Humans
Induced Pluripotent Stem Cells / cytology*
Molecular Imaging
Patch-Clamp Techniques / methods

Substances

Calcium

Grants and funding

P50 NS038377/NS/NINDS NIH HHS/United States