title: A tissue retrieval and postharvest processing regimen for rodent reproductive tissues compatible with long-term storage on the international space station and postflight biospecimen sharing program.
abstract: Collection and processing of tissues to preserve space flight effects from animals after return to Earth is challenging. Specimens must be harvested with minimal time after landing to minimize postflight readaptation alterations in protein expression/translation, posttranslational modifications, and expression, as well as changes in gene expression and tissue histological degradation after euthanasia. We report the development of a widely applicable strategy for determining the window of optimal species-specific and tissue-specific posteuthanasia harvest that can be utilized to integrate into multi-investigator Biospecimen Sharing Programs. We also determined methods for ISS-compatible long-term tissue storage (10 months at -80°C) that yield recovery of high quality mRNA and protein for western analysis after sample return. Our focus was reproductive tissues. The time following euthanasia where tissues could be collected and histological integrity was maintained varied with tissue and species ranging between 1 and 3 hours. RNA quality was preserved in key reproductive tissues fixed in RNAlater up to 40 min after euthanasia. Postfixation processing was also standardized for safe shipment back to our laboratory. Our strategy can be adapted for other tissues under NASA's Biospecimen Sharing Program or similar multi-investigator tissue sharing opportunities.
authors: Department of Molecular & Integrative Physiology, University of Kansas Medical Center, Mail Stop 3050, 3901 Rainbow Boulevard, HLSIC 3098, Kansas City, KS 66160, USA.
authors: Vijayalaxmi
authors: 0000-0001-6339-5570
authors: V
authors: Gupta
authors: Department of Molecular & Integrative Physiology, University of Kansas Medical Center, Mail Stop 3050, 3901 Rainbow Boulevard, HLSIC 3098, Kansas City, KS 66160, USA.
authors: Lesya
authors: L
authors: Holets-Bondar
authors: Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160, USA ; Institute for Reproductive Health and Regenerative Medicine, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors: Katherine F
authors: KF
authors: Roby
authors: Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors: George
authors: G
authors: Enders
authors: Department of Molecular & Integrative Physiology, University of Kansas Medical Center, Mail Stop 3050, 3901 Rainbow Boulevard, HLSIC 3098, Kansas City, KS 66160, USA.
authors: Joseph S
authors: JS
authors: Tash
mesh_terms: D000818:Animals; D005260:Female; D005835:Genitalia / Q000166:cytology / Q000502:physiology*; D005836:Genitalia, Female / Q000166:cytology / Q000502:physiology*; D005849:Gerbillinae; D008297:Male; D008810:Mice, Inbred C57BL; D011309:Preservation, Biological; D055550:Protein Stability; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D013026:Space Flight; D013997:Time Factors; D016707:Tissue Fixation; D020858:Tissue and Organ Harvesting / Q000379:methods*
chemical_list: D011506:Proteins; D012313:RNA
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