{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:19:18Z","timestamp":1785410358228,"version":"3.56.0"},"posted":{"date-parts":[[2025,10,14]]},"group-title":"Cell Biology","reference-count":63,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T00:00:00Z","timestamp":1760400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"award":["R01HD101452"],"award-info":[{"award-number":["R01HD101452"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R21HD107212"],"award-info":[{"award-number":["R21HD107212"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Mammalian spermatogonial stem cells (SSCs) sustain male fertility through continuous self-renewal and differentiation, leading to the production of haploid spermatozoa throughout adulthood. However, SSCs are vulnerable to genotoxic drugs, and patients receiving chemotherapy face a high risk of germline instability and infertility. The molecular mechanisms and cellular pathways that choreograph SSC recovery after chemotherapeutic insult remain unknown. Previously, we identified SPRY4 as an ERK-dependent negative feedback regulator of growth factor signaling that is critical for preservation of stem cell activity in cultured mouse SSCs. Here, we demonstrate that following alkylating agent busulfan (BU)-induced injury in adult mice, germline-specific\n                  <jats:italic>Spry4<\/jats:italic>\n                  gene deletion (\n                  <jats:italic>Spry4<\/jats:italic>\n                  <jats:sup>G-KO<\/jats:sup>\n                  ) reduces stem cell regeneration with an enhanced genotoxic stress response and differentiation with rapidly enhanced nuclear ERK1\/2 activity in undifferentiated (A\n                  <jats:sub>undiff<\/jats:sub>\n                  ) spermatogonia (including SSCs). Genes essential for stem cell maintenance, including\n                  <jats:italic>Id1<\/jats:italic>\n                  and\n                  <jats:italic>Cxcl12<\/jats:italic>\n                  , were dysregulated by loss of\n                  <jats:italic>Spry4<\/jats:italic>\n                  . Furthermore, the MEK1\/2 inhibitor PD0325901, but not mTORC1 inhibitor Rapamycin, was sufficient to promote spermatogonial proliferation in\n                  <jats:italic>Spry4<\/jats:italic>\n                  <jats:sup>G-KO<\/jats:sup>\n                  testis 10 days post-BU treatment. Notably, the restoration of both spermatogonia pool and fertility was delayed in adult\n                  <jats:italic>Spry4<\/jats:italic>\n                  <jats:sup>G-KO<\/jats:sup>\n                  males long-term after injury. In summary, germline-specific deletion of\n                  <jats:italic>Spry4<\/jats:italic>\n                  results in hyper-activation of the MAPK\/ERK pathway in A\n                  <jats:sub>undiff<\/jats:sub>\n                  spermatogonia, reducing spermatogonial genome integrity, unleashing excessive spermatogenesis after germline damage, and ultimately impairing germline regeneration in adult males. Our study indicates an essential role for SPRY4-ERK signaling as a molecular checkpoint in securing SSC recovery upon chemotherapy drug-induced germline damage, revealing how stem cells normally withstand environmental stress.\n                <\/jats:p>","DOI":"10.1101\/2025.10.12.681919","type":"posted-content","created":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T18:00:17Z","timestamp":1760464817000},"source":"Crossref","is-referenced-by-count":0,"title":["Restoration of Spermatogenesis is Dependent on Activation of a SPRY4-ERK Checkpoint Following Germline Stem Cell Damage"],"prefix":"10.64898","author":[{"given":"Ying","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Surgery, Weill Cornell Medicine, New York, NY, USA"},{"name":"Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Ansary Stem Cell Institute, Department of Medicine, Weill Cornell Medicine, New York, NY, 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