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. 2008 Mar 28:4:12.
doi: 10.1186/1746-6148-4-12.

Identification of differentially expressed genes in ileal Peyer's patch of scrapie-infected sheep using RNA arbitrarily primed PCR

Lars Austbø  1 , Andreas Kampmann, Ulf Müller-Ladner, Elena Neumann, Ingrid Olsaker, Grethe Skretting
Affiliations

Identification of differentially expressed genes in ileal Peyer's patch of scrapie-infected sheep using RNA arbitrarily primed PCR

Lars Austbø et al. BMC Vet Res. .

Abstract

Background: In scrapie and prion diseases, the knowledge concerning genes involved in host response during the early infection period in the lymphoid tissues, still remains limited. In the present study, we have examined differential gene expression in ileal Peyer's patches and in laser microdissected follicles of sheep infected with scrapie.

Methods: Ileal Peyer's patches and laser microdissected follicles were of scrapie and control lambs with susceptible genotypes for classical scrapie. Potential regulated genes were found using RNA arbitrarily primed polymerase chain reaction (RAP-PCR) and fingerprinting. The differentially expressed genes were confirmed using real-time RT-PCR.

Results: The expression of three genes (MAPRE3, LOC729073 and DNAJC3), were found to be significantly altered in scrapie infected lambs (P < 0.05).

Conclusion: The three genes have not previously been associated with prion diseases and are interesting as they may reflect biological processes involved in the molecular pathogenesis of prion diseases.

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Figures

Figure 1
Figure 1
Laser-assisted microdissection of follicles in the ileal PP. Nuclei of the tissue sections were stained with hematoxylin. After laser-assisted dissection of each area (a) the samples were collected (b) using an adhesive membrane and transferred to a cap of a 0.5 ml reaction tube and kept at -20°C until RNA extraction. Bar: 200 μm. (c) A section of RAP-PCR fingerprint showing follicles from control animals (lanes 1–2), and follicles from scrapie animals (lanes 3–4). Arrowheads show a band differentially amplified by RAP-PCR indicating an up-regulation in the follicles of the scrapie infected animals.
Figure 2
Figure 2
RAP-PCR fingerprints showing the three genes that were verified as differentially expressed in the ileal PP: a) DNAJC3 amplified using primer Br2, b) LOC729073 amplified using primer Hurra3 and c) MAPRE3 amplified using primer OPN23. The lanes represent ileal PP from control animals (Co) and ileal PP from scrapie animals (Sc). Arrowheads indicate the bands of the differentially expressed genes.
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