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Levels of Acid Sphingomyelinase (ASM) in Caenorhabditis elegans in Microgravity

Annabel K. Gravely

Annabel K. Gravely

University of Toronto SchoolsToronto,
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Gravely, Annabel K.
, 
Alice Vlasov

Alice Vlasov

University of Toronto SchoolsToronto,
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Vlasov, Alice
, 
Amy Freeman

Amy Freeman

University of Toronto SchoolsToronto,
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Freeman, Amy
, 
Kay Wu

Kay Wu

University of Toronto SchoolsToronto,
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Wu, Kay
, 
Nathaniel J. Szewczyk

Nathaniel J. Szewczyk

University of Nottingham, MRC/Arthritis Research UK Centre for Musculoskeletal Ageing Research & National Institute for Health Research (NIHR) Nottingham Biomedical Research Centre, Royal Derby Hospital Centre
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Szewczyk, Nathaniel J.
, 
Robert D’Cruz

Robert D’Cruz

Keenan Research Center for Biomedical Sciences, St. Michael's HospitalToronto,
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D’Cruz, Robert
 and 
Jane Batt

Jane Batt

Keenan Research Center for Biomedical Sciences, St. Michael's HospitalToronto,
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Batt, Jane
  
Jul 20, 2020
Levels of Acid Sphingomyelinase (ASM) in Caenorhabditis elegans in Microgravity's Cover Image
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Article Category: Research Article
Published Online: Jul 20, 2020
Page range: 27 - 36
DOI: https://doi.org/10.2478/gsr-2018-0003
Keywords
© 2018 Annabel K. Gravely et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Figure 1

Type 3 Fluids Mixing Enclosure (FME) Mini-Lab. The FME is 17 cm in length with a total volume of 8.4 mL divided into three 2.8 mL compartments. Volume 1 and 3 contained 2.4 mL of CeMM and Volume 2 contained 2.4 mL of CeMM housing 5,000 C. elegans.
Type 3 Fluids Mixing Enclosure (FME) Mini-Lab. The FME is 17 cm in length with a total volume of 8.4 mL divided into three 2.8 mL compartments. Volume 1 and 3 contained 2.4 mL of CeMM and Volume 2 contained 2.4 mL of CeMM housing 5,000 C. elegans.

Figure 2

Length and cross-sectional area (CSA) of ground and space worms. Representative images of (A) ground worms, and (B) space worms are shown. Live C. elegans appear curled and dead C. elegans assume a straight and thick shape. Only live C. elegans were measured. The space worms (N=40) were longer (C), and had larger CSA (D), compared to the control worms (N=40). Data are mean ± SD. ***p<0.05.
Length and cross-sectional area (CSA) of ground and space worms. Representative images of (A) ground worms, and (B) space worms are shown. Live C. elegans appear curled and dead C. elegans assume a straight and thick shape. Only live C. elegans were measured. The space worms (N=40) were longer (C), and had larger CSA (D), compared to the control worms (N=40). Data are mean ± SD. ***p<0.05.

Figure 3

Histograms of length and cross-sectional area (CSA) of space and control C. elegans. The top panels depict the distribution of lengths of randomly selected control C. elegans (n=40, left panel) and space C. elegans (n=40, right panel). Bottom panels similarly demonstrate the distribution of CSAs of control (left panel) and space (right panel) worms. A broader distribution of both length and CSA is evident in the space worms compared to the control ground worms.
Histograms of length and cross-sectional area (CSA) of space and control C. elegans. The top panels depict the distribution of lengths of randomly selected control C. elegans (n=40, left panel) and space C. elegans (n=40, right panel). Bottom panels similarly demonstrate the distribution of CSAs of control (left panel) and space (right panel) worms. A broader distribution of both length and CSA is evident in the space worms compared to the control ground worms.

Figure 4

Ground and space worm lysates immunoblotted for ASM-1 and ASM-2. The space worms exhibited decreased expression of both ASM-1 and ASM-2, compared to the ground worms (left panel ASM Western blot, right panel graph of quantified fluorescent signal). GAPDH immunostaining demonstrates equivalent protein loading. Data are mean ± SD. ***p<0.05, **p<0.1.
Ground and space worm lysates immunoblotted for ASM-1 and ASM-2. The space worms exhibited decreased expression of both ASM-1 and ASM-2, compared to the ground worms (left panel ASM Western blot, right panel graph of quantified fluorescent signal). GAPDH immunostaining demonstrates equivalent protein loading. Data are mean ± SD. ***p<0.05, **p<0.1.

Timeline of the experiment_ The letter A stands for arrival (at the ISS) and the letter U stands for undock from the ISS_

LocationTime StartTime DurationTemperature (°C)Changes made to the FME
Travel from Toronto to Houston (FedEx)A-25 days2 days2–4None
NanoRacks (Houston)A-23 days12 days2–4None
Travel from Ferry to LaunchA-11 days11 days2–4None
Rocket travel to ISSA-6 hours6 hours2–4None
ISSA6 weeks21–24A+0 days: Unclamp Clamp A, shake gently for 3 seconds U-14 days: Unclamp Clamp B, shake gently for 3 seconds
Travel from Landing to HoustonU4 days0–10None
Travel from Houston to Toronto (FedEx)U+4 days24 hours2–4None
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