Abstract Kinesin and kinesin-like proteins (KLPs) constitute a superfamily of microtubule motor proteins found in all eukaryotic organisms. Members of the kinesin superfamily are known to play important roles in many fundamental cellular and developmental processes. To date, few published studies have reported on the effects of microgravity on kinesin expression. In this paper, we describe the expression pattern and microgravity-sensitive genes of kinesin in rat bone marrow stromal cells cultured in a ground-based rotating bioreactor. The quantity of kinesin under the clinorotation condition was examined by immunoblot analysis with anti-kinesin. Furthermore, the distribution of kinesin at various times during clinorotation was determined by dual immunostaining, using anti-kinesin monoclonal antibody or anti-β-tubulin monoclonal antibody. In terms of kinesin quantity, we found that the ratios of the amounts of clinorotated/stationary KLPs decreased from clinorotation day 5 to day 10, although it increased on days 2 and 3. Immunofluorescence analysis revealed that kinesin in the nucleus was the first to be affected by simulated microgravity, following the kinesin at the periphery that was affected at various times during clinorotation. Real-time RT-PCR analysis of kinesin mRNA expression was performed and led to the identification of 3 microgravity-sensitive kinesin genes: KIF9, KIFC1, and KIF21A. Our results suggest that kinesin has a distinct expression pattern, and the identification of microgravity-sensitive kinesin genes offers insight into fundamental cell biology.
Detection of the quantity of kinesin and microgravity-sensitive kinesin genes in rat bone marrow stromal cells grown in a simulated microgravity environment
Acta Astronautica ; 68 , 11-12 ; 1722-1728
2010-12-21
7 pages
Article (Journal)
Electronic Resource
English
Kinesin , Kinesin-like proteins , Clinorotation , Microgravity-sensitive kinesin genes , Kinesin expression , BMSCs , bone marrow stromal cells , BSA , bovine serum albumin , DMEM , Dulbecco's Modified Eagle's Medium , EDTA , ethylenediaminetetraacetic acid , EGTA , ethylene glycol-bis(3-aminoethyl ether)-<italic>N</italic>,<italic>N</italic>,<italic>N</italic>′,<italic>N</italic>′-tetraacetate , FITC , fluorescein isothiocyanate , GAPDH , glyceraldehyde-3-phosphate dehydrogenase , HEPES , Na-<italic>N</italic>-(2-hydroxy-ethyl) piperazine-<italic>N</italic>′-2-ethanesulfonic acid , KLP , kinesin-like protein , MT , microtubule , MTOC , microtubule-organizing center , PCR , polymerase chain reaction , PBS , phosphate-buffered saline , PIPES , piperazine-<italic>N</italic>′, <italic>N</italic>-bis 2-ethanesulfonic acid , TRITC , tetramethylrhodamine isothiocyanate
Response and adaptation of bone cells to simulated microgravity
Online Contents | 2014
|Response and adaptation of bone cells to simulated microgravity
Elsevier | 2014
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