- We find that, as found for solitary chemosensory cells (SCCs) and brush cells in the airway epithelium, and for tuft cells in the intestine, the transcriptome of TRPM5-expressing microvillous cells indicates that they are likely involved in the inflammatory response elicited by viral infection of the olfactory epithelium.. - We crossed a mouse expressing mCherry in the nuclei of OSNs under control of the OMP promoter (OMP-H2B::mCherry mice) with TRPM5-eGFP transgenic mice [14] (OMP- H2B::mCherry/TRPM5-eGFP mice). - TRPM5-eGFP mice expressed nuclear mCherry driven by the OMP promoter in the intermediate layer of the olfactory epithelium (Fig. - eGFP- cells (Fig. - eGFP- cells both the MVC_eGFP cells and OSN_eGFP+. - RNAseq indicates that MVC_eGFP and OSN_eGFP- are distinct groups of chemosensory cells in the mouse olfactory epithelium. - A total of 1073 Olfr genes were included in the analysis (including pseu- dogenes). - While 9 were expressed at higher levels in the MVC_eGFP population their expression levels was very low (<. - a TRPM5 promoter driven expression of eGFP and OMP promoter driven expression of mCherry in the olfactory epithelium. - MVC_eGFP cells expressed genes involved in the taste transduction path- way as expected for chemosensory epithelial cells of the olfactory epithelium [86]. - In contrast, OSN_eGFP- expressed transcripts for markers of canonical OSNs such as OMP, BBS1 and 2 and proteins involved in the canonical olfactory transduction pathway. - eGFP and OSN_eGFP- cells of this study and ChAT- eGFP MVCs and ChAT-eGFP SCCs profiled in the re- spiratory epithelium in the study of Ualiyeva and co- workers [86]. - OSN_eGFP- cells. - c Volcano plot of all olfactory receptors, demonstrating the large number of enriched olfactory receptors in the OSN_eGFP- population. - at higher levels in MVC_eGFP cells. - MVC_eGFP cells. - was lower in OSN_eGFP+ cells (Figure 4 -figure supple- ment 1 shows the results of RNAseq and Figure 4 -figure supplement 2 summarizes the data). - eGFP+ and OSN_eGFP- cells. - eGFP cells respectively (Figure 4 -figure supplement 4).. - eGFP+ compared to both OSN_eGFP- and MVC_eGFP.. - In situ hybridization chain reaction finds strong TRPM5 mRNA expression in MVC_eGFP cells, but not in the nuclear OSN layer. - Studies with regular in situ hybridization find expres- sion of TRPM5 mRNA in MVCs, but not in the OSN nuclear layer [71, 92]. - Consistent with published results [71, 92] we find strong in situ signal for TRPM5 in MVC_eGFP cells located in the apical layer of the olfactory epi- thelium (Fig. - 5a, asterisks, also see Figure 5 – figure supplement 1 for a 3D rendering) and this signal is absent in MVC_eGFP cells in the TRPM5 knockout (Fig. - In addition, we find sparse TRPM5 in situ labeling in the nuclear OSN layer (Fig. - We performed transcriptional profiling of three chemo- sensory cells in the mouse olfactory epithelium: micro- villous cells (MVC_eGFP) and two types of olfactory sensory neurons: OSN_eGFP+ and OSN_eGFP-. - c Volcano plot of all Olfactory receptors, demonstrating the small number of enriched olfactory receptors in the OSN_eGFP+ population. - GOnet GO term enrichment analysis [70] of all 133 immune genes expressed at higher levels in MVC_eGFP cells compared to OSN_eGFP- (Figure 3 – figure supplement 2) revealed that MVCs express a substantial number of genes involved in the innate im- mune response (GO genes matched this list). - Among the list of in- flammation genes found at significantly higher levels in MVC_eGFP compared to OSN_eGFP- cells we find Il25, an interleukin that is involved in the type 2 inflamma- tory response of TRPM5-expressing epithelial cells in the airway epithelium and the gut [62, 84]. - Also, Il25 ex- pression in the skin leads to disruption of the epithelium and enhances HSV-1 and vaccinia virus replication [38].. - Finally, the innate immune response involves recruitment of macrophages that are known to play a protective role in the olfactory epithelium [6].. - In a similar manner in the olfactory epithelium MVCs respond to ATP, which is involved in activating mucociliary movement by releasing acetylcholine and ac- tivating adjacent sustentacular cells through a. - 5 In situ hybridization chain reaction finds strong TRPM5 mRNA expression in MVC_eGFP cells, but not in the nuclear OSN layer. - Pou2f3 transcript was increased in MVC_eGFP cells compared to OSN_eGFP-. - in the intestine [29]. - 6 Model depicting the role for microvillous cells involvement in the olfactory epithelium innate immune response to viral infection. - 3) MVC_eGFP express numerous transcriptional factors involved in the inhibition of viral replication. - Are GFP-expressing OSNs in the TRPM5-GFP mouse a distinct set of OSNs?. - Transcriptional profiling of single cells in the olfactory epithelium from other laboratories found expression of transcripts for both Tmprss2 and Ace2 in in sustentacu- lar cells and stem cells, and at lower levels in MVCs [7, 24]. - These expression patterns suggest that, like tuft cells in the gut and SCCs and brush cells in the airways, the microvillous cells in the olfactory epithelium are involved in the innate im- mune response to viral infection. - eGFP cells and OSN_eGFP+ cells that constitute a small fraction of the cells in the epithelium, we used FACS to separate the cell populations targeted for RNAseq [3]. - When we moved the OMP-H2B::mCherry/TRPM5-eGFP to HV cages we noticed a decrease in the number of OSN_. - The number of OSN_eGFP+. - Following euthanasia via CO2 inhalation, the olfactory epithelium was immediately removed from the nasal cavity and epithelial tissue was separated from the bone in the turbinates. - org/10.1186/s y.. - performed in situ experiments and litera- ture search and wrote the section on viral infection in the discussion. - Differential distribution of six calcium-binding proteins in the rat olfactory epithelium during postnatal development and adulthood. - Macrophage depletion in the murine olfactory epithelium leads to increased neuronal death and decreased neurogenesis. - https://doi.org/10.1002/cne.21252.. - Non-neuronal expression of SARS-CoV-2 entry genes in the olfactory system suggests mechanisms underlying COVID-19-associated anosmia. - https://doi.org/10.101 6/j.cell . - https://doi.org . - https://doi.org/10.1093/chemse/bjj027.. - Detection of sweet and umami taste in the absence of taste receptor T1r3.. - https://doi.. - https://doi.org/10.1093/chemse/bju015.. - SARS- CoV-2 receptor and entry genes are expressed by sustentacular cells in the human olfactory neuroepithelium. - https://doi.org/10.3389/fncel . - https://doi.org/10.1038/nature05732.. - Microvillous cells in the olfactory epithelium express elements of the solitary chemosensory cell transduction signaling cascade.. - https://doi.org/10.1038/nature1652 7.. - https://doi.org/10.1093/. - Neuregulin1 and ErbB expression in the uninjured and regenerating olfactory mucosa. - https://doi.org/10.1016/j.gep . - https://doi.org/10.1016/j.febslet . - https://doi.org/10.3389/fnagi . - Microvillous cells expressing IP3 receptor type 3 in the olfactory epithelium of mice. - https://doi.org/10.1111/j x.. - https://doi.org/10.1038/s . - https://doi.org/10.1073/pnas . - https://doi.org/10.1016/j.. - https://doi.org/10.1016/j.antiviral . - https://doi.org/10.1016/j.conb . - TRPM5-expressing microvillous cells in the main olfactory epithelium. - Transduction for pheromones in the Main olfactory epithelium is mediated by the Ca2+. - https://doi.org/10.1186/s . - https://doi.org/10.1016/j.wjorl . - https://doi.org a0.. - doi.org/10.1523/JNEUROSCI . - org/10.1016/S . - https://doi.org/10.1146/annurev-immunol . - Cholinergic microvillous cells in the mouse main olfactory epithelium and effect of acetylcholine on olfactory sensory neurons and supporting cells. - https://doi.org/10.1152/jn . - Trpc2-expressing sensory neurons in the Main olfactory epithelium of the mouse. - Trpm5 expression in the olfactory epithelium. - https://doi.org/10.1016/j.mcn . - P/Q type Calcium Channel Cav2.1 defines a unique subset of glomeruli in the mouse olfactory bulb. - Development of solitary chemosensory cells in the distal lung after severe influenza injury. - doi.org/10.1152/ajplung . - https://doi.org/10.. - https://doi.org/10.1073/pna s . - https://doi.org X-6-53.. - Sex separation induces differences in the olfactory sensory receptor repertoires of male and female mice.. - https://doi.org/10.1523/JNEUROSCI . - https://doi.org/10.1038/nature16161.. - Skn- 1a/Pou2f3 is required for the generation of Trpm5-expressing microvillous cells in the mouse main olfactory epithelium. - https://doi.org/10.1016/j.celrep . - https://doi.org/10.1093/nar/gkx1098.. - https://doi.org/10.101 6/S . - https://doi.org/10.1016/j.cell
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