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  • Title: Plasma and intracellular membrane inositol 1,4,5-trisphosphate receptors mediate the Ca(2+) increase associated with the ATP-induced increase in ciliary beat frequency.
    Author: Barrera NP, Morales B, Villalón M.
    Journal: Am J Physiol Cell Physiol; 2004 Oct; 287(4):C1114-24. PubMed ID: 15175223.
    Abstract:
    An increase in intracellular free Ca(2+) concentration ([Ca(2+)](i)) has been shown to be involved in the increase in ciliary beat frequency (CBF) in response to ATP; however, the signaling pathways associated with inositol 1,4,5-trisphosphate (IP(3)) receptor-dependent Ca(2+) mobilization remain unresolved. Using radioimmunoassay techniques, we have demonstrated the appearance of two IP(3) peaks occurring 10 and 60 s after ATP addition, which was strongly correlated with a release of intracellular Ca(2+) from internal stores and an influx of extracellular Ca(2+), respectively. In addition, ATP-dependent Ca(2+) mobilization required protein kinase C (PKC) and Ca(2+)/calmodulin-dependent protein kinase II activation. We found an increase in PKC activity in response to ATP, with a peak at 60 s after ATP addition. Xestospongin C, an IP(3) receptor blocker, significantly diminished both the ATP-induced increase in CBF and the initial transient [Ca(2+)](i) component. ATP addition in the presence of xestospongin C or thapsigargin revealed that the Ca(2+) influx is also dependent on IP(3) receptor activation. Immunofluorescence and confocal microscopic studies showed the presence of IP(3) receptor types 1 and 3 in cultured ciliated cells. Immunogold electron microscopy localized IP(3) receptor type 3 to the nucleus, the endoplasmic reticulum, and, interestingly, the plasma membrane. In contrast, IP(3) receptor type 1 was found exclusively in the nucleus and the endoplasmic reticulum. Our study demonstrates for the first time the presence of IP(3) receptor type 3 in the plasma membrane in ciliated cells and leads us to postulate that the IP(3) receptor can directly trigger Ca(2+) influx in response to ATP.
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