Intercellular Bridge Mediates Ca²⁺ Signals between Micropatterned Cells via IP₃ and Ca²⁺ Diffusion.

ABSTRACT: Intercellular bridges are plasma continuities formed at the end of the cytokinesis process that facilitate intercellular mass transport between the two daughter cells. However, it remains largely unknown how the intercellular bridge mediates Ca²⁺ communication between postmitotic cells. In this work, we utilize BV-2 microglial cells planted on dumbbell-shaped micropatterned assemblies to resolve spatiotemporal characteristics of Ca²⁺ signal transfer over the intercellular bridges. With the use of such micropatterns, considerably longer and more regular intercellular bridges can be obtained than in conventional cell cultures. The initial Ca²⁺ signal is evoked by mechanical stimulation of one of the daughter cells. A considerable time delay is observed between the arrivals of passive Ca²⁺ diffusion and endogenous Ca²⁺ response in the intercellular-bridge-connected cell, indicating two different pathways of the Ca²⁺ communication. Extracellular Ca²⁺ and the paracrine pathway have practically no effect on the endogenous Ca²⁺ response, demonstrated by application of Ca²⁺-free medium, exogenous ATP, and P2Y₁₃ receptor antagonist. In contrast, the endoplasmic reticulum Ca²⁺-ATPase inhibitor thapsigargin and inositol trisphosphate (IP₃) receptor blocker 2-aminoethyl diphenylborate significantly inhibit the endogenous Ca²⁺ increase, which signifies involvement of IP₃-sensitive calcium store release. Notably, passive Ca²⁺ diffusion into the connected cell can clearly be detected when IP₃-sensitive calcium store release is abolished by 2-aminoethyl diphenylborate. Those observations prove that both passive Ca²⁺ diffusion and IP₃-mediated endogenous Ca²⁺ response contribute to the Ca²⁺ increase in intercellular-bridge-connected cells. Moreover, a simulation model agreed well with the experimental observations.

SUBMITTER: Xing F 

PROVIDER: S-EPMC7063411 | biostudies-literature |

REPOSITORIES: biostudies-literature