Other
16 Authors
- Kage H,
- Horie M,
- Zhou B,
- Wang H,
- Marconett CN,
- Borok Z,
- Hung L,
- Offringa IA,
- Wilkinson TA,
- Castaldi A,
- Li F,
- Ji Y,
- Sunohara M,
- Flodby P,
- Liu Y,
- Shen H
| First Author | Horie M | Year | 2020 |
| Journal | Cells | Volume | 9 |
| Issue | 11 | PubMed ID | 33187367 |
| Mgi Jnum | J:329563 | Mgi Id | MGI:6751300 |
| Doi | 10.3390/cells9112460 | Citation | Horie M, et al. (2020) Integrated Single-Cell RNA-Sequencing Analysis of Aquaporin 5-Expressing Mouse Lung Epithelial Cells Identifies GPRC5A as a Novel Validated Type I Cell Surface Marker. Cells 9(11):2460 |
| abstractText | Molecular and functional characterization of alveolar epithelial type I (AT1) cells has been challenging due to difficulty in isolating sufficient numbers of viable cells. Here we performed single-cell RNA-sequencing (scRNA-seq) of tdTomato(+) cells from lungs of AT1 cell-specific Aqp5-Cre-IRES-DsRed (ACID);R26tdTomato reporter mice. Following enzymatic digestion, CD31(-)CD45(-)E-cadherin(+)tdTomato(+) cells were subjected to fluorescence-activated cell sorting (FACS) followed by scRNA-seq. Cell identity was confirmed by immunofluorescence using cell type-specific antibodies. After quality control, 92 cells were analyzed. Most cells expressed 'conventional' AT1 cell markers (Aqp5, Pdpn, Hopx, Ager), with heterogeneous expression within this population. The remaining cells expressed AT2, club, basal or ciliated cell markers. Integration with public datasets identified three robust AT1 cell- and lung-enriched genes, Ager, Rtkn2 and Gprc5a, that were conserved across species. GPRC5A co-localized with HOPX and was not expressed in AT2 or airway cells in mouse, rat and human lung. GPRC5A co-localized with AQP5 but not pro-SPC or CC10 in mouse lung epithelial cell cytospins. We enriched mouse AT1 cells to perform molecular phenotyping using scRNA-seq. Further characterization of putative AT1 cell-enriched genes revealed GPRC5A as a conserved AT1 cell surface marker that may be useful for AT1 cell isolation. |
