{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T13:05:45Z","timestamp":1793106345802,"version":"4.5.0"},"reference-count":84,"publisher":"Oxford University Press (OUP)","issue":"6","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,12,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Purpose<\/jats:title>\n                    <jats:p>Developing protocols for ovarian cortex preservation paves the way for collection of valuable early germ cells from all female individuals, regardless of age and reproductive status. This study aimed to further optimize the microwave-assisted dehydration protocol by characterizing ovarian tissue stress response to drying in the domestic cat model.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods and findings<\/jats:title>\n                    <jats:p>Ovaries from prepubertal cats were dissected using different techniques, exposed to different treatments, and analyzed for ovarian cortex integrity and function after dehydration and rehydration protocols. Using RNA sequencing, we highlighted functions and pathways most affected by dehydration and selected marker genes for quantitative PCR to test different protocol conditions. Switching to a biopsy puncher over our previously used dissection technique introduced the first major advancement in the protocol, improving follicular integrity and transcriptional activity after 10\u00a0min of drying compared to our previous reports. This change also improved the expression of genes related to signaling, membrane transport, and transcription regulation after 10\u00a0min of drying (loss of 87.3% of water), while almost all genes were significantly downregulated after 15\u00a0min of drying (loss of 89.7% of water). Using lucifer yellow as a proxy for trehalose, we indirectly estimated trehalose uptake into follicles after membrane permeabilization with digitonin. Although increased digitonin incubation resulted in potentially higher uptake of trehalose, the associated membrane damage led to compromised follicular integrity and decreased transcriptional activity after dehydration.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>These results expand our knowledge of the ovarian tissue stress response to the dehydration challenge and bring us closer to optimizing the tissue preservation protocol at ambient temperatures.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Summary Sentence<\/jats:title>\n                    <jats:p>This study advances ambient-temperature preservation of ovarian cortex by characterizing tissue stress response to the microwave-assisted dehydration protocol and exploring ways to reduce stress-induced damage in the domestic cat model.<\/jats:p>\n             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