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The findings were verified in rat and human IVD by immunostaining and qRT\u2010PCR. Tamoxifen\u2010administered Pdgfra\n                      <jats:sup>CreERT2<\/jats:sup>\n                      ;R26\n                      <jats:sup>tdTomato<\/jats:sup>\n                      mice were adopted to track platelet\u2010derived growth factor receptor alpha (Pdgfra) positive cells.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Puncture response regions were revealed on day 1 post\u2010puncture, for which oxidative stress emerged as a prominent pathway in a Stress Zone consisting of lipocalin\u20102 (Lcn2)+ annulus fibrosus (AF) cells (AFC), which propagated and migrated into nucleus pulposus (NP), playing a key role in delivering injury signals and triggering pathological processes, including ferroptosis, fibrosis, and immune reactions. In the NP, Collagen 3\u2010high (Col3hi) NP cells (NPC) were another induced population demonstrating a fibrochondrocyte\u2010like phenotype and high epithelial\u2013mesenchymal transition activation, an important pathway involved in tissue fibrosis. Crucially, lineage tracing experiments in Pdgfra\n                      <jats:sup>CreERT2<\/jats:sup>\n                      ;R26\n                      <jats:sup>tdTomat<\/jats:sup>\n                      mice revealed the significant migration and proliferation of Pdgfra+ AFCs from the AF into the NP following puncture. These findings provide direct evidence that both Pdgfra+ AFCs and Col3hi NP cells may contribute to NP fibrosis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Puncture\u2010induced oxidative stress in a stress zone is the primary reaction playing an important role in initiating IVDD. Several puncture\u2010induced cell subpopulations were identified, including Lcn2+ AFC, Col3hi NPC, and Pdgfra+ AFC. Lcn2+ AFC plays a pivotal role in connecting oxidative stress with other pathological processes. Our results clarified the dual origin of Pdgfra+ cells, highlighting the contribution of AF\u2010derived cells to the NP during degeneration and emphasizing the complexity of cellular changes underlying NP fibrosis. Further investigation into the specific contributions of Pdgfra+ cells from different origins to fibrosis is warranted.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Key points<\/jats:title>\n                    <jats:p>\n                      <jats:list list-type=\"bullet\">\n                        <jats:list-item>\n                          <jats:p>Puncture induced oxidative stress in a Stress Zone is the primary reaction in initiating IVDD.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>Puncture induced several IVD cell subpopulations, including Lcn2+ AFC, Col3hi NPC and Pdgfra+ AFC.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>Lcn2+ AFC plays a pivotal role in connecting oxidative stress with other pathological processes.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>Pdgfra+ cells in the NP derived from both Pdgfra+ AFC and Col3hi NPC, highlighting dual origin of NP fibrosis.<\/jats:p>\n                        <\/jats:list-item>\n                      <\/jats:list>\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/ctm2.70370","type":"journal-article","created":{"date-parts":[[2025,6,17]],"date-time":"2025-06-17T08:04:14Z","timestamp":1750147454000},"source":"Crossref","is-referenced-by-count":6,"title":["Single nuclear\u2010spatial transcriptomic sequencing reveals distinct puncture\u2010induced cell subpopulations in the intervertebral disc of a rat model"],"prefix":"10.1002","volume":"15","author":[{"given":"Guoyan","family":"Liang","sequence":"first","affiliation":[{"name":"Department of Spine Surgery Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences) Southern Medical University  Guangzhou P.R. 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