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Temporal RNA Integrity Analysis of Archived Spaceflight Biological Samples

Elizabeth Delgadillo Talburt
Elizabeth Delgadillo Talburt
, 
Alison J. French
Alison J. French
, 
Danielle K. Lopez
Danielle K. Lopez
, 
San-Huei Lai Polo
San-Huei Lai Polo
, 
Valery Boyko
Valery Boyko
, 
Marie T. Dinh
Marie T. Dinh
, 
Jon C. Rask
Jon C. Rask
, 
Helen J. Stewart
Helen J. Stewart
 et 
Kaushik Chakravarty
Kaushik Chakravarty
   | 21 juil. 2020
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Article Category: Research Article
Publié en ligne: 21 juil. 2020
Pages: 44 - 53
DOI: https://doi.org/10.2478/gsr-2018-0009
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© 2018 Elizabeth Delgadillo Talburt et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Quantitative temporal comparison of RNA integrity number (RIN) values of rodent liver tissue samples from scientific payloads returned from the International Space Station from 1991 to 2016. Error bars represent standard error of the mean. Significance within a 99% confidence interval was determined using Tukey's Multiple Comparisons Test. Significant difference from all (p < 0.01); significance from Spacelab Life Sciences 1 (SLS-1), Rodent Research 1 (RR1), and Rodent Research 3 (RR3) (p < 0.01); and significance from SLS-1, Commercial Biomedical Test Module 3 (CBTM-3), and the control (p < 0.01) is indicated by the symbols ¥, †, and ‡, respectively. The sample sizes for SLS-1, CBTM-3, RR1, RR3, and control tissues are n = 4, n = 9, n = 10, n = 20, and n = 12, respectively.
Quantitative temporal comparison of RNA integrity number (RIN) values of rodent liver tissue samples from scientific payloads returned from the International Space Station from 1991 to 2016. Error bars represent standard error of the mean. Significance within a 99% confidence interval was determined using Tukey's Multiple Comparisons Test. Significant difference from all (p < 0.01); significance from Spacelab Life Sciences 1 (SLS-1), Rodent Research 1 (RR1), and Rodent Research 3 (RR3) (p < 0.01); and significance from SLS-1, Commercial Biomedical Test Module 3 (CBTM-3), and the control (p < 0.01) is indicated by the symbols ¥, †, and ‡, respectively. The sample sizes for SLS-1, CBTM-3, RR1, RR3, and control tissues are n = 4, n = 9, n = 10, n = 20, and n = 12, respectively.

Figure 2

Qualitative temporal comparison of RNA depicting the overall quality of each extraction protocol generated using the Agilent Bioanalyzer 2100 System. Extractions were derived from the Spacelab Life Sciences 1 (SLS-1), Commercial Biomedical Test Module 3 (CBTM-3, Rodent Research 1 (RR1), and Rodent Research 3 (RR3) and control tissue.
Qualitative temporal comparison of RNA depicting the overall quality of each extraction protocol generated using the Agilent Bioanalyzer 2100 System. Extractions were derived from the Spacelab Life Sciences 1 (SLS-1), Commercial Biomedical Test Module 3 (CBTM-3, Rodent Research 1 (RR1), and Rodent Research 3 (RR3) and control tissue.

Figure 3

RIN comparison between payloads by treatment group. Payloads divided into subgroups by treatment type – ground, flight, and basal – when applicable. There was no significant difference between treatment groups within any payload. In each subgroup, the payloads SLS-1, CBTM-3, RR1, and RR3 had sample sizes of n = 4, n = 3, n = 5, and n = 10, respectively.
RIN comparison between payloads by treatment group. Payloads divided into subgroups by treatment type – ground, flight, and basal – when applicable. There was no significant difference between treatment groups within any payload. In each subgroup, the payloads SLS-1, CBTM-3, RR1, and RR3 had sample sizes of n = 4, n = 3, n = 5, and n = 10, respectively.

Figure 4

Mean RIN values of RR3 livers compared between fixative methods. Error bars represent SEM and the asterisk indicates a significant difference (p < 0.05, two-tailed t-test with Welch's Correction). For the liquid nitrogen fixed group and the RNAlater fixed group, the sample sizes were n = 12 and n = 8, respectively.
Mean RIN values of RR3 livers compared between fixative methods. Error bars represent SEM and the asterisk indicates a significant difference (p < 0.05, two-tailed t-test with Welch's Correction). For the liquid nitrogen fixed group and the RNAlater fixed group, the sample sizes were n = 12 and n = 8, respectively.

Tissues and protocols used in experimental study.

YearExperimental GroupsPayloadAnimal ModelStorage ContainerFixativeDissection Protocol
1991Ground (N=4)SLS-1RatFoilLN2Rats were dissected upon shuttle return. Rats were sacrificed by decapitation, then dissected immediately. Livers were removed, placed on tinfoil, and frozen on dry ice immediately.
2011Ground (N=3)Flight (N=3)Basal (N=3)CBTM-3MouseCryovialLN2Mice were sacrificed by 4% isoflurane inhalation and dissected within 3–5.5 hours after shuttle landing and snap-frozen in liquid nitrogen.
2014Ground (N=5)Flight (N=5)RR1MouseCryovialLN2Mice were sacrificed on board the ISS, then frozen at −80°C. Mouse carcasses were taken from −80°C freezer and thawed at room temperature for approximately 20 minutes. Livers were taken and either snap-frozen in LN2 or placed into cryovials with RNAlater.
2016Ground (N=6)Flight (N=6)RR3MouseCryovialLN2Mice were sacrificed on board the ISS, then frozen at −80°C. Mouse carcasses were taken from −80°C freezer and thawed at room temperature for approximately 20 minutes. Livers were taken and either snap-frozen in LN2 or placed into cryovials with RNAlater.
2016Ground (N=4)Flight (N=4)RR3MouseCryovialRNAlaterMice were sacrificed on board the ISS, then frozen at −80°C. Mouse carcasses were taken from −80°C freezer and thawed at room temperature for approximately 20 minutes. Livers were taken and either snap-frozen in LN2 or placed into cryovials with RNAlater.
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