Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesis

Abstract Cellular decision-making and tissue homeostasis are governed by transcriptional networks shaped by chromatin accessibility. Using single-nucleus multi-omics, we jointly profile gene expression and chromatin accessibility in 10,335 cells from the Drosophila testis apical tip. This enables in...

Full description

Saved in:
Bibliographic Details
Main Authors: Patrick van Nierop y Sanchez, Pallavi Santhi Sekhar, Kerem Yildirim, Tim Lange, Laura Zoe Kreplin, Vigneshwarr Muruga Boopathy, Stephanie Rosswag de Souza, Kim Dammer, David Ibberson, Qian Wang, Katrin Domsch, Anniek Stokkermans, Shubhanshu Pandey, Petra Kaspar, Rafael Martinez-Gallegos, Xuefan Gao, Aakriti Singh, Natalja Engel, Fillip Port, Michael Boutros, Josephine Bageritz, Ingrid Lohmann
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62046-9
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849234720467976192
author Patrick van Nierop y Sanchez
Pallavi Santhi Sekhar
Kerem Yildirim
Tim Lange
Laura Zoe Kreplin
Vigneshwarr Muruga Boopathy
Stephanie Rosswag de Souza
Kim Dammer
David Ibberson
Qian Wang
Katrin Domsch
Anniek Stokkermans
Shubhanshu Pandey
Petra Kaspar
Rafael Martinez-Gallegos
Xuefan Gao
Aakriti Singh
Natalja Engel
Fillip Port
Michael Boutros
Josephine Bageritz
Ingrid Lohmann
author_facet Patrick van Nierop y Sanchez
Pallavi Santhi Sekhar
Kerem Yildirim
Tim Lange
Laura Zoe Kreplin
Vigneshwarr Muruga Boopathy
Stephanie Rosswag de Souza
Kim Dammer
David Ibberson
Qian Wang
Katrin Domsch
Anniek Stokkermans
Shubhanshu Pandey
Petra Kaspar
Rafael Martinez-Gallegos
Xuefan Gao
Aakriti Singh
Natalja Engel
Fillip Port
Michael Boutros
Josephine Bageritz
Ingrid Lohmann
author_sort Patrick van Nierop y Sanchez
collection DOAJ
description Abstract Cellular decision-making and tissue homeostasis are governed by transcriptional networks shaped by chromatin accessibility. Using single-nucleus multi-omics, we jointly profile gene expression and chromatin accessibility in 10,335 cells from the Drosophila testis apical tip. This enables inference of 147 cell type-specific enhancer-gene regulons using SCENIC + . We functionally validate key transcription factors, including ovo and klumpfuss, known from other stem cell systems but not previously linked to spermatogenesis. CRISPR-mediated knockout reveals their essential roles in germline stem cell regulation, and we provide evidence that they co-regulate shared targets through overlapping enhancer elements. We further uncover a critical role for canonical Wnt signaling, with Pangolin/Tcf activating lineage-specific targets in the germline, soma, and niche. The Pan eRegulon links Wnt activity to cell adhesion, intercellular signaling and germline stem cell maintenance. Together, our study defines the enhancer-driven regulatory landscape of early spermatogenesis and reveals conserved, combinatorial mechanisms of niche-dependent stem cell control.
format Article
id doaj-art-bf2a620753274ea4a2392fce368c8a7f
institution Kabale University
issn 2041-1723
language English
publishDate 2025-07-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-bf2a620753274ea4a2392fce368c8a7f2025-08-20T04:03:03ZengNature PortfolioNature Communications2041-17232025-07-0116111910.1038/s41467-025-62046-9Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesisPatrick van Nierop y Sanchez0Pallavi Santhi Sekhar1Kerem Yildirim2Tim Lange3Laura Zoe Kreplin4Vigneshwarr Muruga Boopathy5Stephanie Rosswag de Souza6Kim Dammer7David Ibberson8Qian Wang9Katrin Domsch10Anniek Stokkermans11Shubhanshu Pandey12Petra Kaspar13Rafael Martinez-Gallegos14Xuefan Gao15Aakriti Singh16Natalja Engel17Fillip Port18Michael Boutros19Josephine Bageritz20Ingrid Lohmann21Heidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceDeep Sequencing Core Facility, BioQuant, Heidelberg UniversityHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHubrecht Institute-KNAWHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Stem Cell Niche HeterogeneityGerman Cancer Research Center (DKFZ), Division of Signaling and Functional Genomics and University of Heidelberg, Department of Cell and Molecular BiologyGerman Cancer Research Center (DKFZ), Division of Signaling and Functional Genomics and University of Heidelberg, Department of Cell and Molecular BiologyGerman Cancer Research Center (DKFZ), Division of Signaling and Functional Genomics and University of Heidelberg, Department of Cell and Molecular BiologyHeidelberg University, Centre for Organismal Studies (COS) Heidelberg, Department of Developmental Biology and Cell Networks—Cluster of ExcellenceAbstract Cellular decision-making and tissue homeostasis are governed by transcriptional networks shaped by chromatin accessibility. Using single-nucleus multi-omics, we jointly profile gene expression and chromatin accessibility in 10,335 cells from the Drosophila testis apical tip. This enables inference of 147 cell type-specific enhancer-gene regulons using SCENIC + . We functionally validate key transcription factors, including ovo and klumpfuss, known from other stem cell systems but not previously linked to spermatogenesis. CRISPR-mediated knockout reveals their essential roles in germline stem cell regulation, and we provide evidence that they co-regulate shared targets through overlapping enhancer elements. We further uncover a critical role for canonical Wnt signaling, with Pangolin/Tcf activating lineage-specific targets in the germline, soma, and niche. The Pan eRegulon links Wnt activity to cell adhesion, intercellular signaling and germline stem cell maintenance. Together, our study defines the enhancer-driven regulatory landscape of early spermatogenesis and reveals conserved, combinatorial mechanisms of niche-dependent stem cell control.https://doi.org/10.1038/s41467-025-62046-9
spellingShingle Patrick van Nierop y Sanchez
Pallavi Santhi Sekhar
Kerem Yildirim
Tim Lange
Laura Zoe Kreplin
Vigneshwarr Muruga Boopathy
Stephanie Rosswag de Souza
Kim Dammer
David Ibberson
Qian Wang
Katrin Domsch
Anniek Stokkermans
Shubhanshu Pandey
Petra Kaspar
Rafael Martinez-Gallegos
Xuefan Gao
Aakriti Singh
Natalja Engel
Fillip Port
Michael Boutros
Josephine Bageritz
Ingrid Lohmann
Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesis
Nature Communications
title Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesis
title_full Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesis
title_fullStr Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesis
title_full_unstemmed Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesis
title_short Dissecting the enhancer gene regulatory network in early Drosophila spermatogenesis
title_sort dissecting the enhancer gene regulatory network in early drosophila spermatogenesis
url https://doi.org/10.1038/s41467-025-62046-9
work_keys_str_mv AT patrickvannieropysanchez dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT pallavisanthisekhar dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT keremyildirim dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT timlange dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT laurazoekreplin dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT vigneshwarrmurugaboopathy dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT stephanierosswagdesouza dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT kimdammer dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT davidibberson dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT qianwang dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT katrindomsch dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT anniekstokkermans dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT shubhanshupandey dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT petrakaspar dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT rafaelmartinezgallegos dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT xuefangao dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT aakritisingh dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT nataljaengel dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT fillipport dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT michaelboutros dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT josephinebageritz dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis
AT ingridlohmann dissectingtheenhancergeneregulatorynetworkinearlydrosophilaspermatogenesis