Measurement and analysis of calcium signaling in heterogeneous cell cultures.
High-content imaging platforms capable of studying kinetic responses at a single-cell level have elevated kinetic recording techniques from labor-intensive low-throughput experiments to potential high-throughput screening assays. We have applied this technology to the investigation of heterogeneous cell cultures derived from primary neural tissue. The neuronal cultures mature into a coupled network and display spontaneous oscillations in intracellular calcium, which can be modified by the addition of pharmacological agents. We have developed algorithms to perform Fourier analysis and quantify both the degree of synchronization and the effects of modulators on the oscillations. Functional and phenotypic experiments can be combined using this approach. We have used post-hoc immunolabeling to identify subpopulations of cells in cocultures and to dissect the calcium responses of these cells from the population response. The combination of these techniques represents a powerful tool for drug discovery.
Richards
Gillian R
GR
The Neuroscience Research Centre, Merck Sharp and Dohme Research Laboratories, Essex, United Kingdom.
Jack
Andrew D
AD
Platts
Amy
A
Simpson
Peter B
PB
D000465:Algorithms; D000818:Animals; D001921:Brain / Q000196:embryology; D020013:Calcium Signaling; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured / Q000737:chemistry*; D002540:Cerebral Cortex / Q000378:metabolism; D002626:Chemistry, Pharmaceutical / Q000379:methods; D015195:Drug Design; D005583:Fourier Analysis; D007700:Kinetics; D009474:Neurons / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D013997:Time Factors