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Increased Expression of Axogenesis-Related Genes and Mossy Fibre Length in Dentate Granule Cells from Adult HuD Overexpressor Mice

Perrone-Bizzozero, Nora I, Tanner, Daniel C, Mounce, Joanna, Bolognani, Federico·2011·ASN Neuro
DOI 10.1042/an20110015PMC PMC3234101
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Figure 1 LCM of DGCs in HuD-Tg mice and control littermates (A, B) DGC from adult control mice (A) do not express HuD but high levels of the protein is expressed in HuD-Tg mice (B). Scale bar = 150 μm. (C–E) H/E (haematoxylin/eosin) staining demonstrating the successful laser capture of cells in the DGC layer.
Figure 1 LCM of DGCs in HuD-Tg mice and control littermates (A, B) DGC from adult control mice (A) do not express HuD but high levels of the protein is expressed in HuD-Tg mice (B). Scale bar = 150 μm. (C–E) H/E (haematoxylin/eosin) staining demonstrating the successful laser capture of cells in the DGC layer.CC BY-NC · PubMed Central
Figure 2 Increased TTR mRNA levels in DGCs of HuD-Tg mice (A) TTR FISH images from WT and HuD-Tg mice. Low magnification taken using a ×4 objective show expression of TTR mRNA in CP in WT and HuD-Tg mice and in the dentate gyrus of HuD-Tg mice. Scale bar = 750 μm. High magnification (×20) images show increase TTR mRNA in DGC of HuD Tg mice. Scale bar = 150 μm. (B) Quantification of TTR mRNA levels in DGCs of WT and HuD-Tg by ISH and qRT–PCR was performed as described in the Materials and methods section. *P<0.05 and ***P<0.001.
Figure 2 Increased TTR mRNA levels in DGCs of HuD-Tg mice (A) TTR FISH images from WT and HuD-Tg mice. Low magnification taken using a ×4 objective show expression of TTR mRNA in CP in WT and HuD-Tg mice and in the dentate gyrus of HuD-Tg mice. Scale bar = 750 μm. High magnification (×20) images show increase TTR mRNA in DGC of HuD Tg mice. Scale bar = 150 μm. (B) Quantification of TTR mRNA levels in DGCs of WT and HuD-Tg by ISH and qRT–PCR was performed as described in the Materials and methods section. *P<0.05 and ***P<0.001.CC BY-NC · PubMed Central
Figure 3 Analyses of 3′-UTR motifs in HuD-DGCs up-regulated mRNAs The frequencies of the different types of ARE motifs, including the consensus motif in the ARED, GRE motif and HuD-binding motifs (see text for details) in the 3′-UTRs of HuD-DGC transcripts (DG-up, black bars) were analysed and compared with all the genes present in the Affymetrix 430 2.0 chip (Affy-chip white bars), and a list of the top 500 DG expressed transcripts that are present in control mice and whose expression levels are not significantly different in HuD-Tg mice (DG-P500, grey bars). Statistical analyses were performed using a one-tail χ2 test: ****P<0.00001, DG-up compared with both DG-P500 and Affy-chip, ***P = 0.0024, DG-up compared with Affy-chip, **P = 0.0093 DG-up compared with Affy-chip and *P = 0.040 compared with DG-P500.
Figure 3 Analyses of 3′-UTR motifs in HuD-DGCs up-regulated mRNAs The frequencies of the different types of ARE motifs, including the consensus motif in the ARED, GRE motif and HuD-binding motifs (see text for details) in the 3′-UTRs of HuD-DGC transcripts (DG-up, black bars) were analysed and compared with all the genes present in the Affymetrix 430 2.0 chip (Affy-chip white bars), and a list of the top 500 DG expressed transcripts that are present in control mice and whose expression levels are not significantly different in HuD-Tg mice (DG-P500, grey bars). Statistical analyses were performed using a one-tail χ2 test: ****P<0.00001, DG-up compared with both DG-P500 and Affy-chip, ***P = 0.0024, DG-up compared with Affy-chip, **P = 0.0093 DG-up compared with Affy-chip and *P = 0.040 compared with DG-P500.CC BY-NC · PubMed Central
Figure 4 GO analysis of up-regulated transcripts GO analyses were performed as indicated in the Materials and methods section. Panels show the observed (blue bars) and expected (red bars) number of genes in the biological process, molecular function and cellular component categories. The top ten significantly enriched categories are presented.
Figure 4 GO analysis of up-regulated transcripts GO analyses were performed as indicated in the Materials and methods section. Panels show the observed (blue bars) and expected (red bars) number of genes in the biological process, molecular function and cellular component categories. The top ten significantly enriched categories are presented.CC BY-NC · PubMed Central
Figure 5 Biological network of genes involved in nervous system development IPA showing interactions of gene products from HuD-Tg overexpressed genes. Proteins that are involved in nervous system development and function and whose transcripts were up-regulated in our dataset are shown in red, whereas transcripts that were absent in our dataset but whose products are involved in nervous system development are shown in blue. Continuous lines represent direct relationships, whereas broken lines represent indirect relationships. The biological activities associated with each protein symbol are indicated in the key box.
Figure 5 Biological network of genes involved in nervous system development IPA showing interactions of gene products from HuD-Tg overexpressed genes. Proteins that are involved in nervous system development and function and whose transcripts were up-regulated in our dataset are shown in red, whereas transcripts that were absent in our dataset but whose products are involved in nervous system development are shown in blue. Continuous lines represent direct relationships, whereas broken lines represent indirect relationships. The biological activities associated with each protein symbol are indicated in the key box.CC BY-NC · PubMed Central
Figure 6 Increased length of the infrapyramidal mossy fibre bundle in HuD-Tg mice (A) Shows the distribution of IPB and SPB bundles in WT and HuD transgenic mice. Mossy fibres were stained using either calbindin antibodies (B) or Timm's stain (C). The IPB length was calculated from the hilus to where the fibres cross the pyramidal cell layer (arrows), and the length of the SPB was measured from the hilus to the apex of the curvature of area CA3 (C). The length of the IPB was normalized to that of the SPB in the same section (D). Scale bar = 300 μm. **P<0.01.
Figure 6 Increased length of the infrapyramidal mossy fibre bundle in HuD-Tg mice (A) Shows the distribution of IPB and SPB bundles in WT and HuD transgenic mice. Mossy fibres were stained using either calbindin antibodies (B) or Timm's stain (C). The IPB length was calculated from the hilus to where the fibres cross the pyramidal cell layer (arrows), and the length of the SPB was measured from the hilus to the apex of the curvature of area CA3 (C). The length of the IPB was normalized to that of the SPB in the same section (D). Scale bar = 300 μm. **P<0.01.CC BY-NC · PubMed Central

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Metadata

Authors
Perrone-Bizzozero, N. I., Tanner, D. C., Mounce, J., & Bolognani, F.
Affiliations
Departmemt of Neurosciences, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA
Venue
ASN Neuro
Publisher
Informa UK Limited
Published
2011-10
Volume
3
Issue
5
ISSN
1759-0914
DOI
10.1042/an20110015
PMC
PMC3234101
Cited by
25
References
3
Language
en
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