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Open Access Highly Accessed Research article

Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate

Neva J Fudge and Karen M Mearow*

Author Affiliations

Division of BioMedical Sciences, Memorial University of Newfoundland, St. John’s, NL, Canada

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BMC Neuroscience 2013, 14:15  doi:10.1186/1471-2202-14-15

Published: 30 January 2013

Additional files

Additional file 1: Table S1:

List of ECM genes that were assessed by microarray analysis. Bold type indicates the genes that were detected as being expressed in either or both populations of DRG neurons used in this study. Genes chosen for further analyses were those that were shown to be detected on >3 arrays. Number of biological replicates (and arrays) ranged from 3–7.

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Additional file 2: Figure S1:

Heat maps of gene expression in microarrays. Representative heat maps of 2 different experiments showing differences in gene expression using either the initial arrays (ORN 13) or the second array series (ORN 13.2).

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Additional file 3: Table S2:

36 genes were expressed by both DRG neuron populations. RNA prepared from IB4+ and IB4- DRG neurons was analysed using small scale oligonucleotide microarrays. The values presented are the mean spot density from 3–7 different arrays determined as outlined in the Methods; SEM is shown in italics. Eight genes (

    underlined
) were highly expressed in both populations. Nine genes (shown in bold) were differentially expressed between the IB4+ and IB4- populations at either t=0 or t=24LN.

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Additional file 4: Table S3:

Microarray analysis showed that 21 genes were differentially expressed between the populations or in response to LN. Relative quantity of gene expression was determined by expressing the normalized mean values obtained from microarray analyses relative to the IB4- levels at t=0 or IB4+ at t=0. Values that indicated differential expression (defined as >1.5 fold increase or <0.6 fold decrease) are

    underlined
. Nine genes (shown in bold) were differentially expressed between the IB4+ and IB4- populations at either t=0 or t=24LN, and were chosen for further study. Statistical significance * p<0.05; + p<0.10.

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Additional file 5: Table S4:

qRT-PCR quantitation of gene expression. Nine genes were subjected to qRT-PCR as described in the text and Methods. The data (means + SEM) are expressed relative to the IB4- t=0 condition.

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Additional file 6: Figure S2:

Densitometric analyses of ICC protein expression for selected proteins in dissociated DRG neurons. Quantitation of ICC staining (using average gray level measurements) was performed as outlined in the Methods. Total dissociated neuronal cultures were analysed comparing IB4+ vs IB4- cells in the same culture wells, as well as across time points or plating experiments. Statistical significance was noted by ANOVA or Students t-Test. Numbers of cells counted are noted within the bars.

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Additional file 7: Figure S3:

Colour composite images of immunostained DRG sections – series 1. Cryosections of adult rat DRGs were subject to immunohistochemistry for selected proteins (red or green as noted), as well as concomitant labeling with the IB4-lectin (blue). The final column of panels presents the merged images. Scale bar – 50 μm.

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Additional file 8: Figure S4:

Colour composite images of immunostained DRG sections – series 2. Cryosections of adult rat DRGs were subject to immunohistochemistry for selected proteins (red or green as noted), as well as concomitant labeling with the IB4-lectin (blue). The final column of panels presents the merged images. Scale bar – 50 μm.

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