Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Reiner, Benjamin C., Crist, Richard C., Borner, Tito, Doyle, Robert P., Hayes, Matthew R., De Jonghe, Bart C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8749158/
https://www.ncbi.nlm.nih.gov/pubmed/34942400
http://dx.doi.org/10.1016/j.molmet.2021.101422
_version_ 1784631168023396352
author Reiner, Benjamin C.
Crist, Richard C.
Borner, Tito
Doyle, Robert P.
Hayes, Matthew R.
De Jonghe, Bart C.
author_facet Reiner, Benjamin C.
Crist, Richard C.
Borner, Tito
Doyle, Robert P.
Hayes, Matthew R.
De Jonghe, Bart C.
author_sort Reiner, Benjamin C.
collection PubMed
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.
format Online
Article
Text
id pubmed-8749158
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-87491582022-01-13 Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment Reiner, Benjamin C. Crist, Richard C. Borner, Tito Doyle, Robert P. Hayes, Matthew R. De Jonghe, Bart C. Mol Metab Brief Communication 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. Elsevier 2021-12-21 /pmc/articles/PMC8749158/ /pubmed/34942400 http://dx.doi.org/10.1016/j.molmet.2021.101422 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Brief Communication
Reiner, Benjamin C.
Crist, Richard C.
Borner, Tito
Doyle, Robert P.
Hayes, Matthew R.
De Jonghe, Bart C.
Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment
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 Brief Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8749158/
https://www.ncbi.nlm.nih.gov/pubmed/34942400
http://dx.doi.org/10.1016/j.molmet.2021.101422
work_keys_str_mv AT reinerbenjaminc singlenucleirnasequencingoftheratapandntsfollowinggdf15treatment
AT cristrichardc singlenucleirnasequencingoftheratapandntsfollowinggdf15treatment
AT bornertito singlenucleirnasequencingoftheratapandntsfollowinggdf15treatment
AT doylerobertp singlenucleirnasequencingoftheratapandntsfollowinggdf15treatment
AT hayesmatthewr singlenucleirnasequencingoftheratapandntsfollowinggdf15treatment
AT dejonghebartc singlenucleirnasequencingoftheratapandntsfollowinggdf15treatment