Identification Of Novel Genes Involved In Zebra Mussel (Dreissena Polymorpha) Underwater Adhesion Mechanism.

Overview

Zebra mussels (Dreissena polymorpha) have invaded North America causing economic and ecologic devastation. The zebra mussel attaches firmly to underwater substrates through a complex system of exocrine glands, byssal threads, and adhesive plaques. In this study, tools were developed and experiments were designed in order to better understand the zebra mussel underwater adhesion mechanisms. A normalized cDNA library of the zebra mussel byssus was constructed with the subtractive suppression hybridization (SSH) ...
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Overview

Zebra mussels (Dreissena polymorpha) have invaded North America causing economic and ecologic devastation. The zebra mussel attaches firmly to underwater substrates through a complex system of exocrine glands, byssal threads, and adhesive plaques. In this study, tools were developed and experiments were designed in order to better understand the zebra mussel underwater adhesion mechanisms. A normalized cDNA library of the zebra mussel byssus was constructed with the subtractive suppression hybridization (SSH) technique. 750 non-redundant expressed sequence tags (ESTs) were obtained from the library. A cDNA microarray was developed with the PCR products of 716 ESTs selected from the cDNA library. The newly developed cDNA microarray was successfully used to compare between two groups of mussels with different byssogenenic activities. Between the two groups, 16 genes were differentially expressed with statistical significance. The results were validated by the quantitative PCR (qPCR). To follow up on genes that are either up or downregulated along the course of byssogenesis, a microarray time-course experiment was designed. Samples were collected at seven time intervals; 12 hours, 1 day, 2 days, 3 days, 4 days, 7 days, and 21 days after severing the byssal threads in the treatment group. The numbers of differentially expressed genes identified at these time points were 13, 13, 20, 17, 16, 20, and 29 respectively. An additional experiment was designed to identify the differentially expressed genes in response to the changes of byssogenesis status and environmental factors, including temperature, current velocity, and dissolved oxygen levels, as well as the status of byssogenesis on the expression of foot-unique genes. The expression profiles of 18 genes were found to be altered by two experimental factors, while 117 genes had differentially expressed profiles in response to only one experimental factor. The numbers of the genes modulated by byssogenesis status, temperature, dissolved oxygen, and current velocity were 59, 27, 26, and 9, respectively. Seven genes identified by the time-course experiment and four genes identified by factorial analysis assay were validated by qPCR with the results consistent to microarray results. Detected by RNA fluorescent in situ hybridization (FISH), both genes BG20_A01 and BG97/192_B06 were found to be expressed within the thread-forming glandular cells. The genes BG15_F03 and BG16_H05 were expressed by stem-forming gland cells and plaque-forming gland cells, respectively. A full length gene in the microarray, homologous to insect defensin A, was cloned from the zebra mussel foot. The analysis of the D. polymorpha defensin (Dpd) suggested that the Dpd is homologous to the insect defensin A. The expression of Dpd in hemocytes was found to be inducible by the stimulation of lipopolysaccharides (LPS), peptidoglycan (PGN), and zymosan (ZYM) by using qPCR. The mature recombinant Dpd showed inhibition activity against four stains of Gram-negative bacteria. Dpd was present not only in the foot, but also in a number of other tissues. Interestingly, the expression levels of Dpd during the early stage of byssogenesis were mostly higher than the non-byssogenesis status, which suggested that the activities of Dpd were associated with byssogenesis.
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Product Details

  • ISBN-13: 9781243714336
  • Publisher: BiblioLabsII
  • Publication date: 9/8/2011
  • Pages: 228
  • Product dimensions: 7.44 (w) x 9.69 (h) x 0.48 (d)

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