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Temporal expression of nitric oxide synthase in Ilyanassa obsoleta using an Ilyanassa-specific NOS antibody.

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Allison Deal Weaver (Creator)
Institution
The University of North Carolina at Greensboro (UNCG )
Web Site: http://library.uncg.edu/
Advisor
Mark Hens

Abstract: In the mollusc Ilyanassa obsoleta, metamorphosis is a key event in its life history that is closely regulated by positive and negative modulators. Previous research has determined that nitric oxide (NO) and nitric oxide synthase (NOS) maintain the larval state up to competency. RT-PCR of NOS transcripts, NADPH-diaphorase staining, and immunohistochemistry have all confirmed the presence of NOS during larval development and metamorphosis. However, these assays have given conflicting results on when and where NOS is produced. I developed an Ilyanassa-specific NOS antibody that was used in western blot analysis of larval extracts to determine temporal expression of the NOS protein. In situ hybridization was also done to localize NOS transcripts. My Ilyanassa-specific antibody recognized NOS, but the most prominently recognized epitope, a 66 kDa polypeptide, is likely to be a degradation product of NOS. In situ hybridization on competent larvae provided no meaningful data. My studies of temporal expression of the protein confirmed the previous RT-PCR results which demonstrated a constant level of NOS during larval development and a decrease in NOS during metamorphosis. These results were unlike those of previous NADPH-diaphorase histochemistry and suggest regulation of NOS enzymatic activity. My results indicate that metamorphosis in Ilyanassa may be controlled by the regulation of the quantity of NOS present as well as regulation of the level of NOS enzymatic activity.

Additional Information

Publication
Thesis
Language: English
Date: 2009
Keywords
Ilyanassa obsoleta, Metamorphosis, Mollusc, Nitric oxide synthase
Subjects
Nassarius obsoletus $x Research.
Snails $x Research.
Nitric-oxide synthase.
Nitric oxide $x Metabolism.
Metamorphosis.