Histamine-induced Ca2+ entry in human astrocytoma U373 MG cells: evidence for involvement of store-operated channels.

Abstract:

:Glial and glia-derived cells express a variety of receptors for neurotransmitters and hormones, the majority of which evoke both Ca(2+) release from intracellular stores and Ca(2+) entry across the plasma membrane. We investigated the links between histamine H(1) receptor activation, Ca(2+) release from intracellular stores and Ca(2+) influx in human astrocytoma U373 MG cells. Histamine, through a H(1) receptor-mediated effect, evoked an increase in cytoplasmic free calcium concentration ([Ca(2+)](i)) that occurred in two phases: an initial, transient, increase owing to Ca(2+) mobilization from intracellular pools, and a second, sustained increase dependent on both Ca(2+) influx and continuous receptor occupancy. The characteristics of histamine-induced increases in [Ca(2+)](i) were similar to the capacitative entry evoked by emptying of the Ca(2+) stores with thapsigargine, and different from that observed when Ca(2+) influx was activated with OAG (1-oleoyl-2-acetyl-sn-glycerol), a diacylglycerol (DAG) analog. OAG application or increased endogenous DAG, resulting from DAG kinase inhibition, reduced the histamine-induced response. Furthermore, activation of the DAG target, protein kinase C (PKC), by TPA (12-O-tetradecanoyl 4beta-phorbol 13alpha-acetate) resulted in inhibition of the histamine-induced Ca(2+) response, an action prevented by PKC inhibitors. By using reverse transcriptase-polymerase chain reaction analysis, mRNAs for transient receptor potential channels (TRPCs) 1, 4, and 6 as well as for STIM1 (stromal-interacting molecule) and Orai1 were found to be expressed in the U373 MG cells, and confocal microscopy using specific antibodies revealed the presence of the corresponding proteins. Therefore, TRPCs may be candidate proteins forming store-operated channels in the U373 MG cell line. Further, our results confirm the involvement of PKC in the regulation of H(1) receptor-induced responses and point out to the existence of a feedback mechanism acting via PKC to limit the increase in [Ca(2+)](i).

journal_name

J Neurosci Res

authors

Barajas M,Andrade A,Hernandez-Hernandez O,Felix R,Arias-Montaño JA

doi

10.1002/jnr.21784

subject

Has Abstract

pub_date

2008-11-15 00:00:00

pages

3456-68

issue

15

eissn

0360-4012

issn

1097-4547

journal_volume

86

pub_type

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