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
| Autor: | Katherine F. Roby, Joseph S. Tash, George C. Enders, Vijayalaxmi Gupta, Lesya Holets-Bondar |
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| Rok vydání: | 2015 |
| Předmět: |
Male
Pathology medicine.medical_specialty Time Factors Tissue Fixation Biospecimen Article Subject Preservation Biological lcsh:Medicine Biology Bioinformatics General Biochemistry Genetics and Molecular Biology Protein expression Protein stability medicine Animals Genitalia General Immunology and Microbiology Protein Stability Genitalia.female lcsh:R Proteins Genitalia Female General Medicine Space Flight Mice Inbred C57BL Regimen Tissue and Organ Harvesting Postharvest RNA Female Gerbillinae Research Article |
| Zdroj: | BioMed Research International BioMed Research International, Vol 2015 (2015) |
| ISSN: | 2314-6141 2314-6133 |
| DOI: | 10.1155/2015/475935 |
| Popis: | 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 RNAlaterup 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. |
| Databáze: | OpenAIRE |
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