Simultaneous somatic patch-pipette recording of a single astrocyte to evoke voltage-gated calcium currents, and Ca2+ imaging, were used to study the spatial and temporal profiles of depolarization-induced changes in intracellular Ca2+ ([Ca2+]i) in the processes of cultured rat cortical astrocytes existing as pairs. Transient Ca2+ changes locked to depolarization were observed as microdomains in the processes of the astrocyte pairs, and the responses were more pronounced in the adjoining astrocyte. Considering the functional significance of higher concentrations of glutamate observed in certain pathological conditions, Ca2+ transients were recorded following pretreatment of cells with glutamate (500 μm for 20 min). This showed distance-dependent incremental scaling and attenuation in the presence of the metabotropic glutamate receptor (mGluR) antagonist, α-methyl(4-carboxy-phenyl) glycine (MCPG). Estimation of local Ca2+ diffusion coefficients in the astrocytic processes indicated higher values in the adjoining astrocyte of the glutamate pretreated group. Intracellular heparin introduced into the depolarized astrocyte did not affect the Ca2+ transients in the heparin-loaded astrocyte but attenuated the [Ca2+]i responses in the adjoining astrocyte, suggesting that inositol 1,4,5 triphosphate (IP3) may be the transfer signal. The uncoupling agent, 1-octanol, attenuated the [Ca2+] i responses in both the control and glutamate pretreated astrocytes, indicating the role of gap junctional communication. Our studies indicate that individual astrocytes have distinct functional domains, and that the glutamate-induced alterations in Ca2+ signaling involve a sequence of intra- and intercellular steps in which phospholipase C (PLC), IP3, internal Ca2+ stores, VGCC and gap junction channels appear to play an important role.
- Functional microdomains
- Intracellular and intercellular calcium signaling
- Voltage-gated calcium channels
ASJC Scopus subject areas
- Cellular and Molecular Neuroscience