Studies of Ca2+-ATPase involvement in the gravity-directed calcium current and polar axis alignment of germinating Ceratopteris richardii spores
by Bushart, Thomas James, Ph.D., THE UNIVERSITY OF TEXAS AT AUSTIN, 2007, 101 pages; 3271392

Abstract:

All organisms have been subjected to and have evolved with the ubiquitous force of gravity, and most exhibit the ability to sense and respond to this stimulus. To simplify an investigation of the molecular components of a cell's gravity response, this dissertation employs the single-celled spores of the fern Ceratopteris richardii. These spores have a polar calcium flux that is determined by the gravity vector, but an understanding of what the molecular components driving this flux are and how they influence subsequent developmental processes is lacking. Of the possible molecular components, available literature pointed to Ca2+-ATPase transporters as an obvious key participant and so they were selected as the main molecule of investigation.

Our results describe the first cloned Ca2+-ATPase from C. richardii, CrACA1. CrACA1 has high similarity to known plant Ca 2+-ATPases, specifically plasma membrane (PM) Ca2+-ATPases from Arabidopsis, and exhibits in vivo Ca 2+-ATPase activity. An improved method for the statistical analysis and presentation of qualitative RT-PCR data was employed. The RNA, as well as the protein, of CrACA1 is present during the polarity fixation window which supported the need for further analyses of the role of Ca2+-ATPases.

Our results showing that Ca2+-ATPase inhibitors significantly alter the gravity-directed calcium flux of spores are consistent with previous work but offer valuable new insights. The spore PM Ca2+-ATPases have large impacts on the calcium flux and rhizoid growth but no appreciable impact on polar axis alignment. The results on endomembrane-type Ca 2+-ATPases make it clear that this class of pumps has major roles in both axis alignment and tip growth; rhizoid growth is inhibited but alignment to the gravity vector is improved.

The updated model for gravity perception responses in C. richardii spores places a strong emphasis on calcium channels and Ca 2+-ATPases working in concert to result in a bottom-localized calcium pool to align the polar axis with hints of store-operated calcium mobilization. The work presented represents an increase in our knowledge of one way a single cell can respond to the force of gravity, offering testable hypotheses to further refine gravity perception models incorporating calcium localization.

 
AdviserStanley J. Roux, Jr.
SchoolTHE UNIVERSITY OF TEXAS AT AUSTIN
SourceDAI/B 68-06, p. , Oct 2007
Source TypeDissertation
SubjectsMolecular biology; Cellular biology; Plant Physiology Biology
Publication Number3271392
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