A new approach to inhibiting astrocytic IP3-induced intracellular calcium increase in an astrocyte-neuron co-culture system.
Astrocytes exhibit dynamic Ca2+ mobilization, such as Ca2+ wave and Ca2+ oscillation, via an inositol 1,4,5-triphosphate-induced Ca2+ release (IICR)-dependent mechanism. The physiological functions of astrocytic Ca2+ mobilization, however, are poorly understood. To investigate this issue, we created a plasmid encoding an enhanced green fluorescent protein-tagged inositol 1,4,5-triphosphate absorbent protein and expressed it in cultured astrocytes. Expression of this protein inhibited both IICR and the Ca2+ wave in cultured astrocytes. By combining this method to the single cell electroporation technique, we were able to inhibit IICR specifically in astrocytes in an astrocyte-neuron co-culture system. Our approach provides a useful tool for direct examination of the physiological role of astrocytic Ca2+ signaling on neuronal function.
Saheki
Yasunori
Y
Department of Physiology, Graduate School of Medicine and Dentistry, Okayama University, 2-5-1 Shikata, Okayama 700-8558, Japan.
Li
Sheng-Tian
ST
Matsushita
Masayuki
M
Wu
Yu-Mei
YM
Cai
Wei-Hua
WH
Wei
Fan-Yan
FY
Lu
Yun-Fei
YF
Moriwaki
Akiyoshi
A
Tomizawa
Kazuhito
K
Matsui
Hideki
H
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism; D001921:Brain / Q000166:cytology; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects*; D002450:Cell Communication; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D004347:Drug Interactions; D018274:Electroporation / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005110:Extracellular Space / Q000187:drug effects; D016257:Fura-2 / Q000378:metabolism; D006023:Glycoproteins; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology*; D053496:Inositol 1,4,5-Trisphosphate Receptors; D051379:Mice; D009154:Mutation; D009474:Neurons / Q000502:physiology*; D011189:Potassium Chloride / Q000494:pharmacology; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology
D015220:Calcium Channels; D004791:Enzyme Inhibitors; D006023:Glycoproteins; D053496:Inositol 1,4,5-Trisphosphate Receptors; D018160:Receptors, Cytoplasmic and Nuclear; D049452:Green Fluorescent Proteins; D011189:Potassium Chloride; D019284:Thapsigargin; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate; D002118:Calcium; D016257:Fura-2