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NanoCAGE analysis of the mouse olfactory epithelium identifies the expression of vomeronasal receptors and of proximal LINE elements.

Pascarella G, Lazarevic D, Plessy C, Bertin N, Akalin A, Vlachouli C, Simone R, Faulkner GJ, Zucchelli S, Kawai J, Daub CO, Hayashizaki Y, Lenhard B, Carninci P, Gustincich S - Front Cell Neurosci (2014)

Bottom Line: These loci also show a massive expression of Long Interspersed Nuclear Elements (LINEs).We have validated the expression of selected receptors detected by nanoCAGE with in situ hybridization, RT-PCR and qRT-PCR.This work extends the repertory of receptors capable of sensing chemical signals in the MOE, suggesting intriguing interplays between MOE and VNO for pheromone processing and positioning transcribed LINEs as candidate regulatory RNAs for VRs expression.

View Article: PubMed Central - PubMed

Affiliation: Area of Neuroscience, International School for Advanced Studies (SISSA) Trieste, Italy ; RIKEN Yokohama Institute, Center for Life Science Technologies, Division of Genomic Technologies Tsurumi-ku, Yokohama, Japan.

ABSTRACT
By coupling laser capture microdissection to nanoCAGE technology and next-generation sequencing we have identified the genome-wide collection of active promoters in the mouse Main Olfactory Epithelium (MOE). Transcription start sites (TSSs) for the large majority of Olfactory Receptors (ORs) have been previously mapped increasing our understanding of their promoter architecture. Here we show that in our nanoCAGE libraries of the mouse MOE we detect a large number of tags mapped in loci hosting Type-1 and Type-2 Vomeronasal Receptors genes (V1Rs and V2Rs). These loci also show a massive expression of Long Interspersed Nuclear Elements (LINEs). We have validated the expression of selected receptors detected by nanoCAGE with in situ hybridization, RT-PCR and qRT-PCR. This work extends the repertory of receptors capable of sensing chemical signals in the MOE, suggesting intriguing interplays between MOE and VNO for pheromone processing and positioning transcribed LINEs as candidate regulatory RNAs for VRs expression.

No MeSH data available.


Related in: MedlinePlus

In situ hybridization with Vmn2r26 riboprobes on serial sections of the MOE. (A,B) The Vmn2r26 antisense riboprobe hybridizes with a high number of cells throughout the MOE, mostly located in the basal and medial layers. (C) The Vmn2r26 antisense riboprobe specifically stains a high number of cells throughout the VNO. (D–F) Panels (A–C) merged with stained nuclei (DAPI). Scale bars: (A,B,D,E) 50 μm; (C,F) 60 μm.
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Figure 4: In situ hybridization with Vmn2r26 riboprobes on serial sections of the MOE. (A,B) The Vmn2r26 antisense riboprobe hybridizes with a high number of cells throughout the MOE, mostly located in the basal and medial layers. (C) The Vmn2r26 antisense riboprobe specifically stains a high number of cells throughout the VNO. (D–F) Panels (A–C) merged with stained nuclei (DAPI). Scale bars: (A,B,D,E) 50 μm; (C,F) 60 μm.

Mentions: For the V2Rs family we synthesized riboprobes for Vmn2r26 and Vmn2r69. Due to the high level of homology among V2Rs genes, the antisense probe for Vmn2r26 was able to hybridize to Vmn2r19, Vmn2r23, and Vmn2r24, sharing a homology rate ≥80%. All these V2Rs presented non-repeats mapping TCs in close proximity to the annotated RefSeq 5′ end. The riboprobe for Vmn2r69 was specific for this receptor. Vmn2r26 antisense riboprobe distinguished numerous cells in the middle layer as well as a group of cells located in the basal layer of the MOE; all of them displayed morphological features remarkably similar to those of the OSNs. Vmn2r26+ cells were mainly localized in the central/dorsal turbinates of the MOE (Figures 4A,B,D,E).


NanoCAGE analysis of the mouse olfactory epithelium identifies the expression of vomeronasal receptors and of proximal LINE elements.

Pascarella G, Lazarevic D, Plessy C, Bertin N, Akalin A, Vlachouli C, Simone R, Faulkner GJ, Zucchelli S, Kawai J, Daub CO, Hayashizaki Y, Lenhard B, Carninci P, Gustincich S - Front Cell Neurosci (2014)

In situ hybridization with Vmn2r26 riboprobes on serial sections of the MOE. (A,B) The Vmn2r26 antisense riboprobe hybridizes with a high number of cells throughout the MOE, mostly located in the basal and medial layers. (C) The Vmn2r26 antisense riboprobe specifically stains a high number of cells throughout the VNO. (D–F) Panels (A–C) merged with stained nuclei (DAPI). Scale bars: (A,B,D,E) 50 μm; (C,F) 60 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3927265&req=5

Figure 4: In situ hybridization with Vmn2r26 riboprobes on serial sections of the MOE. (A,B) The Vmn2r26 antisense riboprobe hybridizes with a high number of cells throughout the MOE, mostly located in the basal and medial layers. (C) The Vmn2r26 antisense riboprobe specifically stains a high number of cells throughout the VNO. (D–F) Panels (A–C) merged with stained nuclei (DAPI). Scale bars: (A,B,D,E) 50 μm; (C,F) 60 μm.
Mentions: For the V2Rs family we synthesized riboprobes for Vmn2r26 and Vmn2r69. Due to the high level of homology among V2Rs genes, the antisense probe for Vmn2r26 was able to hybridize to Vmn2r19, Vmn2r23, and Vmn2r24, sharing a homology rate ≥80%. All these V2Rs presented non-repeats mapping TCs in close proximity to the annotated RefSeq 5′ end. The riboprobe for Vmn2r69 was specific for this receptor. Vmn2r26 antisense riboprobe distinguished numerous cells in the middle layer as well as a group of cells located in the basal layer of the MOE; all of them displayed morphological features remarkably similar to those of the OSNs. Vmn2r26+ cells were mainly localized in the central/dorsal turbinates of the MOE (Figures 4A,B,D,E).

Bottom Line: These loci also show a massive expression of Long Interspersed Nuclear Elements (LINEs).We have validated the expression of selected receptors detected by nanoCAGE with in situ hybridization, RT-PCR and qRT-PCR.This work extends the repertory of receptors capable of sensing chemical signals in the MOE, suggesting intriguing interplays between MOE and VNO for pheromone processing and positioning transcribed LINEs as candidate regulatory RNAs for VRs expression.

View Article: PubMed Central - PubMed

Affiliation: Area of Neuroscience, International School for Advanced Studies (SISSA) Trieste, Italy ; RIKEN Yokohama Institute, Center for Life Science Technologies, Division of Genomic Technologies Tsurumi-ku, Yokohama, Japan.

ABSTRACT
By coupling laser capture microdissection to nanoCAGE technology and next-generation sequencing we have identified the genome-wide collection of active promoters in the mouse Main Olfactory Epithelium (MOE). Transcription start sites (TSSs) for the large majority of Olfactory Receptors (ORs) have been previously mapped increasing our understanding of their promoter architecture. Here we show that in our nanoCAGE libraries of the mouse MOE we detect a large number of tags mapped in loci hosting Type-1 and Type-2 Vomeronasal Receptors genes (V1Rs and V2Rs). These loci also show a massive expression of Long Interspersed Nuclear Elements (LINEs). We have validated the expression of selected receptors detected by nanoCAGE with in situ hybridization, RT-PCR and qRT-PCR. This work extends the repertory of receptors capable of sensing chemical signals in the MOE, suggesting intriguing interplays between MOE and VNO for pheromone processing and positioning transcribed LINEs as candidate regulatory RNAs for VRs expression.

No MeSH data available.


Related in: MedlinePlus