Document Detail

Preparatory tests for immunodetection of microtubules in protoplasts during IML-2.
MedLine Citation:
PMID:  11541198     Owner:  NASA     Status:  MEDLINE    
Preparatory tests to improve methods for detection of cortical microtubules (cMTs) in rapeseed protoplasts for the IML-2 experiment "TRANSFORM" were undertaken. This study is based on the results obtained in the "PROTO" experiment onboard the shuttle Discovery during an 8-day IML-1 flight in 1992. The use of free-floating protoplasts on IML-1 made it technically impossible for the astronauts to remove the glutaraldehyde fixative during the orbital period resulting in high background fluorescence which made it very difficult to detect MTs. In order to avoid this on the IML-2 mission, protoplasts will be immobilized in alginate beads. The effect of variable concentrations and fixation periods for two different fixatives on the preservation of cMTs was tested. Best results were obtained using 3.5% paraformaldehyde (PFA) for 1 hour, but 1% glutaraldehyde (GA) also gave acceptable results. The effect of low temperatures on microtubule depolymerization was also examined as freshly isolated protoplasts have to be kept at 5 degrees C for up to 20 hours pre-launch and before activation of Biorack. Only slight changes in cMT-appearance were observed at 4 degrees C indicating a minor depolymerization compared to the cMTs in non-chilled protoplasts.
E B Skagen; O S Rasmussen; T H Iversen
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Microgravity quarterly : MGQ     Volume:  4     ISSN:  0958-5036     ISO Abbreviation:  Microgravity Q     Publication Date:  1994 Apr 
Date Detail:
Created Date:  1999-03-06     Completed Date:  1999-03-06     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  100971563     Medline TA:  Microgravity Q     Country:  ITALY    
Other Details:
Languages:  eng     Pagination:  83-91     Citation Subset:  S    
Department of Botany, University of Trondheim, Norway.
Space Flight Mission:
Flight Experiment; STS-42 Shuttle Project; STS-65 Shuttle Project; manned; short duration
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MeSH Terms
Brassica / cytology,  physiology
Cell Culture Techniques / methods
Cells, Cultured
Cold Temperature
Fixatives / chemistry
Formaldehyde / chemistry
Glutaral / chemistry
Hypocotyl / cytology,  physiology,  ultrastructure
Immunohistochemistry / methods*
Microtubules / physiology,  ultrastructure*
Polymers / chemistry
Protoplasts / cytology*,  physiology,  ultrastructure
Space Flight*
Tissue Fixation / methods*
Reg. No./Substance:
0/Alginates; 0/Fixatives; 0/Polymers; 111-30-8/Glutaral; 30525-89-4/paraform; 50-00-0/Formaldehyde

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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