Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment
Objective: Growth differentiation factor 15 (GDF15) is known to play a role in feeding, nausea, and body weight, with action through the GFRAL-RET receptor complex in the area postrema (AP) and nucleus tractus solitarius (NTS). To further elucidate the underlying cell type-specific molecular mechani...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-02-01
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| Series: | Molecular Metabolism |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2212877821002805 |
| _version_ | 1798031275753209856 |
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| author | Benjamin C. Reiner Richard C. Crist Tito Borner Robert P. Doyle Matthew R. Hayes Bart C. De Jonghe |
| author_facet | Benjamin C. Reiner Richard C. Crist Tito Borner Robert P. Doyle Matthew R. Hayes Bart C. De Jonghe |
| author_sort | Benjamin C. Reiner |
| collection | DOAJ |
| description | Objective: Growth differentiation factor 15 (GDF15) is known to play a role in feeding, nausea, and body weight, with action through the GFRAL-RET receptor complex in the area postrema (AP) and nucleus tractus solitarius (NTS). To further elucidate the underlying cell type-specific molecular mechanisms downstream of GDF15 signaling, we used a single nuclei RNA sequencing (snRNAseq) approach to profile AP and NTS cellular subtype-specific transcriptomes after systemic GDF15 treatment. Methods: AP and NTS micropunches were used for snRNAseq from Sprague Dawley rats 6 h following GDF15 or saline injection, and Seurat was used to identify cellular subtypes and cell type-specific alterations in gene expression that were due to the direct and secondary effects of systemic GDF15 treatment. Results: Using the transcriptome profile of ∼35,000 individual AP/NTS nuclei, we identified 19 transcriptomically distinct cellular subtypes, including a single population Gfral and Ret positive excitatory neurons, representing the primary site of action for GDF15. A total of ∼600 cell type-specific differential expression events were identified in neurons and glia, including the identification of transcriptome alterations specific to the direct effects of GDF15 in the Gfral-Ret positive excitatory neurons and shared transcriptome alterations across neuronal and glial cell types. Downstream analyses identified shared and cell type-specific alterations in signaling pathways and upstream regulatory mechanisms of the observed transcriptome alterations. Conclusions: These data provide a considerable advance in our understanding of AP and NTS cell type-specific molecular mechanisms associated with GDF15 signaling. The identified cellular subtype-specific regulatory mechanism and signaling pathways likely represent important targets for future pharmacotherapies. |
| first_indexed | 2024-04-11T19:54:51Z |
| format | Article |
| id | doaj.art-6f6cbabc2b6a49a58d1c61626c1bfb87 |
| institution | Directory Open Access Journal |
| issn | 2212-8778 |
| language | English |
| last_indexed | 2024-04-11T19:54:51Z |
| publishDate | 2022-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Metabolism |
| spelling | doaj.art-6f6cbabc2b6a49a58d1c61626c1bfb872022-12-22T04:06:12ZengElsevierMolecular Metabolism2212-87782022-02-0156101422Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatmentBenjamin C. Reiner0Richard C. Crist1Tito Borner2Robert P. Doyle3Matthew R. Hayes4Bart C. De Jonghe5Department of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA; Corresponding author. TRL Building, 125 South 31st Street, Philadelphia, PA 19104, USA.Department of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USADepartment of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Biobehavioral Health Sciences, University of Pennsylvania, Philadelphia, PA 19104, USASyracuse University, Department of Chemistry, 111 College Place, Syracuse, NY 13244, USADepartment of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Biobehavioral Health Sciences, University of Pennsylvania, Philadelphia, PA 19104, USADepartment of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Biobehavioral Health Sciences, University of Pennsylvania, Philadelphia, PA 19104, USA; Corresponding author. TRL Building, 125 South 31st Street, Philadelphia, PA 19104, USA.Objective: Growth differentiation factor 15 (GDF15) is known to play a role in feeding, nausea, and body weight, with action through the GFRAL-RET receptor complex in the area postrema (AP) and nucleus tractus solitarius (NTS). To further elucidate the underlying cell type-specific molecular mechanisms downstream of GDF15 signaling, we used a single nuclei RNA sequencing (snRNAseq) approach to profile AP and NTS cellular subtype-specific transcriptomes after systemic GDF15 treatment. Methods: AP and NTS micropunches were used for snRNAseq from Sprague Dawley rats 6 h following GDF15 or saline injection, and Seurat was used to identify cellular subtypes and cell type-specific alterations in gene expression that were due to the direct and secondary effects of systemic GDF15 treatment. Results: Using the transcriptome profile of ∼35,000 individual AP/NTS nuclei, we identified 19 transcriptomically distinct cellular subtypes, including a single population Gfral and Ret positive excitatory neurons, representing the primary site of action for GDF15. A total of ∼600 cell type-specific differential expression events were identified in neurons and glia, including the identification of transcriptome alterations specific to the direct effects of GDF15 in the Gfral-Ret positive excitatory neurons and shared transcriptome alterations across neuronal and glial cell types. Downstream analyses identified shared and cell type-specific alterations in signaling pathways and upstream regulatory mechanisms of the observed transcriptome alterations. Conclusions: These data provide a considerable advance in our understanding of AP and NTS cell type-specific molecular mechanisms associated with GDF15 signaling. The identified cellular subtype-specific regulatory mechanism and signaling pathways likely represent important targets for future pharmacotherapies.http://www.sciencedirect.com/science/article/pii/S2212877821002805Area postremaNucleus of the solitary tractGDF15GFRALRET |
| spellingShingle | Benjamin C. Reiner Richard C. Crist Tito Borner Robert P. Doyle Matthew R. Hayes Bart C. De Jonghe Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment Molecular Metabolism Area postrema Nucleus of the solitary tract GDF15 GFRAL RET |
| title | Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment |
| title_full | Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment |
| title_fullStr | Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment |
| title_full_unstemmed | Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment |
| title_short | Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment |
| title_sort | single nuclei rna sequencing of the rat ap and nts following gdf15 treatment |
| topic | Area postrema Nucleus of the solitary tract GDF15 GFRAL RET |
| url | http://www.sciencedirect.com/science/article/pii/S2212877821002805 |
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