Single-Cell Transcriptomic Analysis of Rat Urethral Tissue Following Simulated Childbirth Injury and Microenergy Acoustic Pulse Therapy
4 affiliations
- 1Knuppe Molecular Urology Laboratory, Department of Urology, School of Medicine, University of California, San Francisco, CA 94143, USA
- 2Department of Urology, Department of Interventional Medicine, and Guangdong Provincial Key Laboratory of Biomedical Imaging, The Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai 519000, China
- 3Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
- 4Andrology Center, Peking University First Hospital, Beijing 100034, China
Abstract
Microenergy acoustic pulse (MAP) therapy has been associated with functional and structural improvement after childbirth-related urethral injury, but its multicellular effects remain incompletely defined. We explored treatment- and injury-associated differences in the urethral microenvironment using single-cell RNA sequencing. Eight female Sprague–Dawley rats were assigned to sham, vaginal balloon dilation plus ovariectomy (VBDO), VBDO plus beta-aminopropionitrile (BAPN), or VBDO plus BAPN and MAP (MAP) groups, with one 24-week-old and one 48-week-old rat per group. Urethral tissues were analyzed for cell composition, differential gene expression, Gene Ontology enrichment, stem/progenitor features, senescence-associated secretory phenotype activity, and CellChat-inferred intercellular communication. A balanced single-cell dataset comprising 23,128 cells was used for downstream comparative analyses and resolved 19 annotated cell-type/state categories. Across the injury- and age-associated experimental conditions, epithelial, vascular, stromal, immune, and muscle compartments showed distinct transcriptional patterns, whereas MAP-treated samples showed a distinct cellular distribution; a separate histological cohort showed higher urethral muscle content. MAP-treated samples also showed differences in CellChat-inferred signaling activity in selected extracellular-matrix, adhesion, vascular, developmental, and neural/neuromuscular pathways, including fibronectin 1 (FN1), collagen (COLLAGEN), angiopoietin (ANGPT), vascular endothelial growth factor (VEGF), Wingless/Integrated (WNT), Notch (NOTCH), reelin (RELN), Slit guidance ligand (SLIT), neural cell adhesion molecule (NCAM), and neurexin (NRXN). Exploratory enrichment analysis identified treatment-associated differences in translation, oxidative metabolism, stress adaptation, synaptic-organization processes, progenitor-associated transcriptional states, and senescence-related programs. The treatment-split marker manifold approximation and projection (UMAPs) are descriptive and do not provide evidence of increased muscle-cell number; the reported difference in muscle-associated staining content derives from a separate animal-level histological analysis. Overall, the findings are descriptive and hypothesis-generating and require validation in independently replicated cohorts.
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BibTeX
@article{lin2026singlecell,
title = {{Single-Cell Transcriptomic Analysis of Rat Urethral Tissue Following Simulated Childbirth Injury and Microenergy Acoustic Pulse Therapy}},
author = {Guiting Lin and Liangyu Zhao and Emily Xing and Tian Bai and Hao Li and Yuan Tang and Thomas Cao and Guifang Wang and Yuxin Tang and Tom F. Lue},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = nov,
doi = {10.3390/ijms27219833},
url = {https://www.mdpi.com/1422-0067/27/21/9833},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Single-Cell Transcriptomic Analysis of Rat Urethral Tissue Following Simulated Childbirth Injury and Microenergy Acoustic Pulse Therapy
AU - Guiting Lin
AU - Liangyu Zhao
AU - Emily Xing
AU - Tian Bai
AU - Hao Li
AU - Yuan Tang
AU - Thomas Cao
AU - Guifang Wang
AU - Yuxin Tang
AU - Tom F. Lue
PY - 2026
DA - 2026/11/03
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27219833
UR - https://www.mdpi.com/1422-0067/27/21/9833
AB - Microenergy acoustic pulse (MAP) therapy has been associated with functional and structural improvement after childbirth-related urethral injury, but its multicellular effects remain incompletely defined. We explored treatment- and injury-associated differences in the urethral microenvironment using single-cell RNA sequencing. Eight female Sprague–Dawley rats were assigned to sham, vaginal balloon dilation plus ovariectomy (VBDO), VBDO plus beta-aminopropionitrile (BAPN), or VBDO plus BAPN and MAP (MAP) groups, with one 24-week-old and one 48-week-old rat per group. Urethral tissues were analyzed for cell composition, differential gene expression, Gene Ontology enrichment, stem/progenitor features, senescence-associated secretory phenotype activity, and CellChat-inferred intercellular communication. A balanced single-cell dataset comprising 23,128 cells was used for downstream comparative analyses and resolved 19 annotated cell-type/state categories. Across the injury- and age-associated experimental conditions, epithelial, vascular, stromal, immune, and muscle compartments showed distinct transcriptional patterns, whereas MAP-treated samples showed a distinct cellular distribution; a separate histological cohort showed higher urethral muscle content. MAP-treated samples also showed differences in CellChat-inferred signaling activity in selected extracellular-matrix, adhesion, vascular, developmental, and neural/neuromuscular pathways, including fibronectin 1 (FN1), collagen (COLLAGEN), angiopoietin (ANGPT), vascular endothelial growth factor (VEGF), Wingless/Integrated (WNT), Notch (NOTCH), reelin (RELN), Slit guidance ligand (SLIT), neural cell adhesion molecule (NCAM), and neurexin (NRXN). Exploratory enrichment analysis identified treatment-associated differences in translation, oxidative metabolism, stress adaptation, synaptic-organization processes, progenitor-associated transcriptional states, and senescence-related programs. The treatment-split marker manifold approximation and projection (UMAPs) are descriptive and do not provide evidence of increased muscle-cell number; the reported difference in muscle-associated staining content derives from a separate animal-level histological analysis. Overall, the findings are descriptive and hypothesis-generating and require validation in independently replicated cohorts.
SN - 1422-0067
ER -