The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genes

Abstract The conserved miR-183/96/182 cluster (miR-183C) is expressed in both corneal resident myeloid cells (CRMCs) and sensory nerves (CSN) and modulates corneal immune/inflammatory responses. To uncover cell type-specific roles of miR-183C in CRMC and CSN and their contributions to corneal physio...

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Main Authors: Naman Gupta, Mallika Somayajulu, Katherine Gurdziel, Giovanni LoGrasso, Haidy Aziz, Rita Rosati, Sharon McClellan, Ahalya Pitchaikannu, Manoranjan Santra, Muhammed Farooq Abdul Shukkur, Paul Stemmer, Linda D. Hazlett, Shunbin Xu
Format: Article
Language:English
Published: Nature Portfolio 2024-04-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-024-58403-1
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author Naman Gupta
Mallika Somayajulu
Katherine Gurdziel
Giovanni LoGrasso
Haidy Aziz
Rita Rosati
Sharon McClellan
Ahalya Pitchaikannu
Manoranjan Santra
Muhammed Farooq Abdul Shukkur
Paul Stemmer
Linda D. Hazlett
Shunbin Xu
author_facet Naman Gupta
Mallika Somayajulu
Katherine Gurdziel
Giovanni LoGrasso
Haidy Aziz
Rita Rosati
Sharon McClellan
Ahalya Pitchaikannu
Manoranjan Santra
Muhammed Farooq Abdul Shukkur
Paul Stemmer
Linda D. Hazlett
Shunbin Xu
author_sort Naman Gupta
collection DOAJ
description Abstract The conserved miR-183/96/182 cluster (miR-183C) is expressed in both corneal resident myeloid cells (CRMCs) and sensory nerves (CSN) and modulates corneal immune/inflammatory responses. To uncover cell type-specific roles of miR-183C in CRMC and CSN and their contributions to corneal physiology, myeloid-specific miR-183C conditional knockout (MS-CKO), and sensory nerve-specific CKO (SNS-CKO) mice were produced and characterized in comparison to the conventional miR-183C KO. Immunofluorescence and confocal microscopy of flatmount corneas, corneal sensitivity, and tear volume assays were performed in young adult naïve mice; 3′ RNA sequencing (Seq) and proteomics in the trigeminal ganglion (TG), cornea and CRMCs. Our results showed that, similar to conventional KO mice, the numbers of CRMCs were increased in both MS-CKO and SNS-CKO vs age- and sex-matched WT control littermates, suggesting intrinsic and extrinsic regulations of miR-183C on CRMCs. The number of CRMCs was increased in male vs female MS-CKO mice, suggesting sex-dependent regulation of miR-183C on CRMCs. In the miR-183C KO and SNS-CKO, but not the MS-CKO mice, CSN density was decreased in the epithelial layer of the cornea, but not the stromal layer. Functionally, corneal sensitivity and basal tear volume were reduced in the KO and SNS-CKO, but not the MS-CKO mice. Tear volume in males is consistently higher than female WT mice. Bioinformatic analyses of the transcriptomes revealed a series of cell-type specific target genes of miR-183C in TG sensory neurons and CRMCs. Our data elucidate that miR-183C imposes intrinsic and extrinsic regulation on the establishment and function of CSN and CRMCs by cell-specific target genes. miR-183C modulates corneal sensitivity and tear production through its regulation of corneal sensory innervation.
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spelling doaj-art-cbf9b527a8844be3b0042541dca45a862025-08-20T03:27:18ZengNature PortfolioScientific Reports2045-23222024-04-0114112310.1038/s41598-024-58403-1The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genesNaman Gupta0Mallika Somayajulu1Katherine Gurdziel2Giovanni LoGrasso3Haidy Aziz4Rita Rosati5Sharon McClellan6Ahalya Pitchaikannu7Manoranjan Santra8Muhammed Farooq Abdul Shukkur9Paul Stemmer10Linda D. Hazlett11Shunbin Xu12Department of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityGenome Sciences Core, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversitySchool of Biological Sciences, Wayne State UniversityInstitute of Environmental Health Sciences, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityInstitute of Environmental Health Sciences, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityDepartment of Ophthalmology, Visual and Anatomical Sciences, School of Medicine, Wayne State UniversityAbstract The conserved miR-183/96/182 cluster (miR-183C) is expressed in both corneal resident myeloid cells (CRMCs) and sensory nerves (CSN) and modulates corneal immune/inflammatory responses. To uncover cell type-specific roles of miR-183C in CRMC and CSN and their contributions to corneal physiology, myeloid-specific miR-183C conditional knockout (MS-CKO), and sensory nerve-specific CKO (SNS-CKO) mice were produced and characterized in comparison to the conventional miR-183C KO. Immunofluorescence and confocal microscopy of flatmount corneas, corneal sensitivity, and tear volume assays were performed in young adult naïve mice; 3′ RNA sequencing (Seq) and proteomics in the trigeminal ganglion (TG), cornea and CRMCs. Our results showed that, similar to conventional KO mice, the numbers of CRMCs were increased in both MS-CKO and SNS-CKO vs age- and sex-matched WT control littermates, suggesting intrinsic and extrinsic regulations of miR-183C on CRMCs. The number of CRMCs was increased in male vs female MS-CKO mice, suggesting sex-dependent regulation of miR-183C on CRMCs. In the miR-183C KO and SNS-CKO, but not the MS-CKO mice, CSN density was decreased in the epithelial layer of the cornea, but not the stromal layer. Functionally, corneal sensitivity and basal tear volume were reduced in the KO and SNS-CKO, but not the MS-CKO mice. Tear volume in males is consistently higher than female WT mice. Bioinformatic analyses of the transcriptomes revealed a series of cell-type specific target genes of miR-183C in TG sensory neurons and CRMCs. Our data elucidate that miR-183C imposes intrinsic and extrinsic regulation on the establishment and function of CSN and CRMCs by cell-specific target genes. miR-183C modulates corneal sensitivity and tear production through its regulation of corneal sensory innervation.https://doi.org/10.1038/s41598-024-58403-1
spellingShingle Naman Gupta
Mallika Somayajulu
Katherine Gurdziel
Giovanni LoGrasso
Haidy Aziz
Rita Rosati
Sharon McClellan
Ahalya Pitchaikannu
Manoranjan Santra
Muhammed Farooq Abdul Shukkur
Paul Stemmer
Linda D. Hazlett
Shunbin Xu
The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genes
Scientific Reports
title The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genes
title_full The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genes
title_fullStr The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genes
title_full_unstemmed The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genes
title_short The miR-183/96/182 cluster regulates sensory innervation, resident myeloid cells and functions of the cornea through cell type-specific target genes
title_sort mir 183 96 182 cluster regulates sensory innervation resident myeloid cells and functions of the cornea through cell type specific target genes
url https://doi.org/10.1038/s41598-024-58403-1
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