Analysis of ATP-Induced Ca 2+ Responses at Single Cell Level in Retinal Pigment Epithelium Monolayers

Adv Exp Med Biol. 2019;1185:525-530. doi: 10.1007/978-3-030-27378-1_86.

Abstract

Calcium is one of the most important second messengers in cells and thus involved in a variety of physiological processes. In retinal pigment epithelium (RPE), Ca2+ and its ATP-dependent signaling pathways play important roles in the retina maintenance functions. Changes in intracellular Ca2+ concentration can be measured from living cells by Ca2+ imaging. Combining these measurements with quantitative analysis of Ca2+ response properties enables studies of signaling pathways affecting RPE functions. However, robust tools for response analysis from large cell populations are lacking. We developed MATLAB-based analysis tools for single cell level Ca2+ response data recorded from large fields of intact RPE monolayers. The analysis revealed significant heterogeneity in ATP-induced Ca2+ responses inside cell populations regarding magnitude and response kinetics. Further analysis including response grouping and parameter correlations allowed us to characterize the populations at the level of single cells.

Keywords: ATP-induced Ca2+ response; Ca2+ imaging; Human embryonic stem cells; Image analysis; Retinal pigment epithelium.

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Calcium Signaling*
  • Cells, Cultured
  • Humans
  • Retinal Pigment Epithelium / cytology*

Substances

  • Adenosine Triphosphate