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.2010 May 28:11:341.
doi: 10.1186/1471-2164-11-341.

Factorial microarray analysis of zebra mussel (Dreissena polymorpha: Dreissenidae, Bivalvia) adhesion

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Factorial microarray analysis of zebra mussel (Dreissena polymorpha: Dreissenidae, Bivalvia) adhesion

Wei Xu et al. BMC Genomics..

Abstract

Background: The zebra mussel (Dreissena polymorpha) has been well known for its expertise in attaching to substances under the water. Studies in past decades on this underwater adhesion focused on the adhesive protein isolated from the byssogenesis apparatus of the zebra mussel. However, the mechanism of the initiation, maintenance, and determination of the attachment process remains largely unknown.

Results: In this study, we used a zebra mussel cDNA microarray previously developed in our lab and a factorial analysis to identify the genes that were involved in response to the changes of four factors: temperature (Factor A), current velocity (Factor B), dissolved oxygen (Factor C), and byssogenesis status (Factor D). Twenty probes in the microarray were found to be modified by one of the factors. The transcription products of four selected genes, DPFP-BG20_A01, EGP-BG97/192_B06, EGP-BG13_G05, and NH-BG17_C09 were unique to the zebra mussel foot based on the results of quantitative reverse transcription PCR (qRT-PCR). The expression profiles of these four genes under the attachment and non-attachment were also confirmed by qRT-PCR and the result is accordant to that from microarray assay. The in situ hybridization with the RNA probes of two identified genes DPFP-BG20_A01 and EGP-BG97/192_B06 indicated that both of them were expressed by a type of exocrine gland cell located in the middle part of the zebra mussel foot.

Conclusions: The results of this study suggested that the changes of D. polymorpha byssogenesis status and the environmental factors can dramatically affect the expression profiles of the genes unique to the foot. It turns out that the factorial design and analysis of the microarray experiment is a reliable method to identify the influence of multiple factors on the expression profiles of the probesets in the microarray; therein it provides a powerful tool to reveal the mechanism of zebra mussel underwater attachment.

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Figures

Figure 1
Figure 1
Comparison of the primary structures of the excretory gland peptide encoding genes whose expression profiles can be modified by byssogenic activity. The comparison was performed by using ClustalW multiple sequences alignment. The protein product encoded by the EST BG97/192_B06 is homologous to an excretory protein isolated from a filarial nematodeLitomosoides sigmodontiswhile the rest of the EGPs encoded by selected ESTs have a salivary gland peptide from blacklegged tick (Ixodes scapularis) as their homologue. The nucleotides were colored with black when more than 50% nucleotides in this locus were identical. The gap between the nucleotides were labeled as "-".
Figure 2
Figure 2
The similarity of the EGP encoding genes demonstrated by multiple sequences alignment indicating the similarity. The three EGP genes are all differentially expressed under the change of water temperature. The EST BG27_B08 is homologous to salivary gland peptide identified from the western black-legged tick (Ixodes pacificus). The other two probesets are all homologous to the blacklegged tick (Ixodes scapularis). The nucleotides were colored with black when more than 50% nucleotides in this locus were identical. The gap between the nucleotides were labeled as "-".
Figure 3
Figure 3
The distribution of the mRNA products of the selected genes within zebra mussel tissues. The (+) indicated the existence of the gene in the tissue; and the (-) suggested no detected transcripts in the tissue. For each gene, the lowest (+) sample was used as control and the relative expression levels of the gene in other tissues were calculated by using 2-ΔΔCtmodel.
Figure 4
Figure 4
The qRT-PCR results demonstrated the relative expression levels of the gene during the byssiogenesis. The byssogenesis and non-byssogenesis samples were treated as described in materials and methods, and the non-byssogenesis sample was used as control with value 1. The amount of the transcripts of the gene was calculated by using 2-ΔΔCtmodel. The comparisons of the gene expression levels between byssogenesis and non-byssogenesis and detached sample were made at 0, 12 hours, 1 day, 2 days, and 3 days post-treatment.
Figure 5
Figure 5
The distribution of the zebra mussel byssus gland cells in mussel foot. The zebra mussel foot sections are stained by H&E method. The sections are made along the longitude axis as demonstrated by the paralleled dash lines across the foot. Arrows with dashed line indicate the position of the three major byssal glands embeded in the mussel's foot. a: A longitudinal section in the root of the mussel's foot. Arrows point to the stem-forming glandular cells (S). b: The section of middle area of the foot. The light purple cells along the epethelial surface of the foot were thread-forming cells which were demostrated by arrows and the letter T. c: The tip of the foot containing the deep purple stained plaque-forming gland cells that were labled as P.
Figure 6
Figure 6
Thein situexpression of the gene DPFP-BG20_A01 and EGP-BG97/192_B06 in zebra mussel foot tissue. The synthesized antisense RNA of DPFP-BG20_A01 was labeled with Alexa 488 dye (green) while the complementary RNA for EGP-BG97/192_B06 was labeled with Alexa 594 (red). The foot was cut along longitude as shown by the two paralleled dash lines across the foot. The positions of the byssus glands were labeled on the foot. The dash lines connected between slides and foot indicated the position of the three slides on the foot. a: The root area of zebra mussel foot including stem-forming gland; b: The middle region of the mussel foot containing thread-forming gland; c: The tip section of the foot including plaque-forming gland. T: Thread-forming gland cells.
Figure 7
Figure 7
The identified probesets withP< 0.05 were hierarchically clustered based on their expression profiles under the effects of the four factors. One hundred and seventeen genes with differentially expressed profiles under the effect of at least one factor were used in this analysis. The logarithmic values of the ratios of two levels in each factor are indicated by different colors. Green and red color stands for up- and down-regulation, respectively. Black encodes no signifcant changes. (A) Hierarchical average linkage clustering. Ten clusters have been created with each of them labeled on the side of the heatmap. (B) The ten clusters represent the genes with the expressions induced by the change of the four factors.
Figure 8
Figure 8
The numbers of genes whose expression profiles are modified by single factor or multiple factors. The values in the ellipse are the numbers of the probes which were modified by the factor with the same color. For example, the sum of the numbers in red ellipse is 19+1+1+6 = 27 meaning that 27 probes were found modulated by the factor A. The underline(s) under each number suggested the amount and types of factors could regulate its expression profiles. For instance, the number 19 with a read underline means the expression profiles of the 19 genes can only be affected by factor A while the value 1 with a red and blue lines suggests that one gene was regulated by both Factor A and Factor B.
Figure 9
Figure 9
Effects of temperature, dissolved oxygen, current velocity, and byssogenesis status on gene expression in the zebra mussel foot using cDNA microarray analysis. The figure displays the different treatment combination selected as per the loop design approach. Each arrow represents one microarray hybridization. The start of the arrow stands for the sample labelled with dye Alexa 647 while the end point of an arrow represents the sample labelled with dye Alexa 555. L stands for low temperature (4°C) while R stands for a higher temperature (22°C); S stands for static water while F means flow water stirred by magnetic stirring bars; H represents the low dissolved oxygen level (5 mg/L) while the N represents the normal dissolved oxygen level (10 mg/L); A and D stand for attachment and detachment status, respectively.
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References

    1. Hebert PDN, Muncaster BW, Mackie GL. Ecological and genetic studies on Dreissena polymorpha (Pallas): a new mollusc in the Great Lakes. Can J Fish Aquat Sci. 1989;46:1587–1591.
    1. Dreissena polymorpha. USGS nonindigenous aquatic species database.http://nas.er.usgs.gov/queries/FactSheet.asp?speciesID=5
    1. Nalepa TF, Schloesser DW. Zebra mussels: biology, impact and control. Boca Raton, Florida: Lewis Publisher; 1993.
    1. New-York-Sea-Grant. Book Dreissena polymorpha information review (Editor ed.^eds.) 1. Vol. 5. City: New York Sea Grant; 1994. Dreissena polymorpha information review; pp. 14–15.
    1. New-York-Sea-Grant. Book "Agency Activities." Dreissena! 3. Vol. 5. City: New York Sea Grant; 1994. "Agency Activities." Dreissena! pp. 1–2.

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