Molecular and functional profiling of histamine receptor-mediated calcium ion signals in different cell lines

Anal Biochem. 2015 Oct 1:486:96-101. doi: 10.1016/j.ab.2015.06.037. Epub 2015 Jul 4.

Abstract

Calcium ions (Ca(2+)) play a pivotal role in cellular physiology. Often Ca(2+)-dependent processes are studied in commonly available cell lines. To induce Ca(2+) signals on demand, cells may need to be equipped with additional proteins. A prominent group of membrane proteins evoking Ca(2+) signals are G-protein coupled receptors (GPCRs). These proteins register external signals such as photons, odorants, and neurotransmitters and convey ligand recognition into cellular responses, one of which is Ca(2+) signaling. To avoid receptor cross-talk or cross-activation with introduced proteins, the repertoire of cell-endogenous receptors must be known. Here we examined the presence of histamine receptors in six cell lines frequently used as hosts to study cellular signaling processes. In a concentration-dependent manner, histamine caused a rise in intracellular Ca(2+) in HeLa, HEK 293, and COS-1 cells. The concentration for half-maximal activation (EC50) was in the low micromolar range. In individual cells, transient Ca(2+) signals and Ca(2+) oscillations were uncovered. The results show that (i) HeLa, HEK 293, and COS-1 cells express sufficient amounts of endogenous receptors to study cellular Ca(2+) signaling processes directly and (ii) these cell lines are suitable for calibrating Ca(2+) biosensors in situ based on histamine receptor evoked responses.

Keywords: Biosensors; Cellular signaling; GPCR; Oscillation; Second messenger.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism
  • Calcium Signaling* / drug effects
  • Cell Line
  • Dose-Response Relationship, Drug
  • Gene Expression Regulation / drug effects
  • Histamine / pharmacology
  • Humans
  • Intracellular Space / drug effects
  • Intracellular Space / metabolism
  • Receptors, Histamine / genetics
  • Receptors, Histamine / metabolism*
  • Single-Cell Analysis

Substances

  • Receptors, Histamine
  • Histamine
  • Calcium