Increases in intracellular calcium via activation of potentially multiple phospholipase C isozymes in mouse olfactory neurons
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Type of Work14 pages
Citation of Original PublicationSteven A. Szebenyi, Tatsuya Ogura, Aaron Sathyanesan, Abdullah K. AlMatrouk, Justin Chang and Weihong Lin, Increases in intracellular calcium via activation of potentially multiple phospholipase C isozymes in mouse olfactory neurons, Front. Cell. Neurosci., DOI: https://doi.org/10.3389/fncel.2014.00336
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Phospholipase C (PLC) and internal Ca²⁺ stores are involved in a variety of cellular functions. However, our understanding of PLC in mammalian olfactory sensory neurons (OSNs) is generally limited to its controversial role in odor transduction. Here we employed single-cell Ca²⁺ imaging and molecular approaches to investigate PLC-mediated Ca²⁺ responses and its isozyme gene transcript expression. We found that the pan-PLC activator m-3M3FBS (25 μM) induces intracellular Ca²⁺ increases in vast majority of isolated mouse OSNs tested. Both the response amplitude and percent responding cells depend on m-3M3FBS concentrations. In contrast, the inactive analog o-3M3FBS fails to induce Ca2+ responses. The m-3M3FBS-induced Ca²⁺ increase is blocked by the PLC inhibitor U73122, while its inactive analog U73433 has no effect. Removal of extracellular Ca²⁺ does not change significantly the m-3M3FBS-induced Ca²⁺ response amplitude. Additionally, in the absence of external Ca²⁺, we found that a subset of OSNs respond to an odorant mixture with small Ca²⁺ increases, which are significantly suppressed by U73122. Furthermore, using reverse transcription polymerase chain reaction and real-time quantitative polymerase chain reaction, we found that multiple PLC isozyme gene transcripts are expressed in olfactory turbinate tissue in various levels. Using RNA in situ hybridization analysis, we further show expression of β4, γ1, γ2 gene transcripts in OSNs. Taken together, our results establish that PLC isozymes are potent enzymes for mobilizing intracellular Ca²⁺ in mouse OSNs and provide molecular insight for PLC isozymes-mediated complex cell signaling and regulation in the peripheral olfactory epithelium.
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