Positive correlation between relative mRNA expression of PRNP and SPRN in cerebral and cerebellar cortex of sheep.

SPRN is an interesting new member of the PRNP family because of its sequence homology with the hydrophobic region of PRNP, its expression in brain tissue and its PrP-like properties in functional experiments on Prnp(0/0) mice. In this study, we investigated by quantitative real-time PCR the relative mRNA expression levels of SPRN and PRNP in sheep cerebrum and cerebellum and the mutual relationship between these expression levels. Analysis of PRNP and SPRN mRNA expression levels in 45 cerebral cortex and 47 cerebellar cortex samples showed that the PRNP expression level was significantly higher (p<0.05) in cerebellum than in cerebrum, while no significant difference was detected for SPRN between these tissues. The expression level varied clearly more in cerebrum than in cerebellum for both genes tested, and the variation was larger for SPRN than for PRNP in both types of brain tissue. Remarkably, the mRNA expression levels of PRNP and SPRN showed a highly significant positive correlation in both cerebrum (p<0.0001) and cerebellum (p<0.001). This positive correlation might indicate co-regulation between these genes. Further investigation on the causal nature of this correlation may provide new insights into prion pathogenesis.

Characterization of the genomic region containing the Shadow of Prion Protein (SPRN) gene in sheep.

BACKGROUND: TSEs are a group of fatal neurodegenerative diseases occurring in man and animals. They are caused by prions, alternatively folded forms of the endogenous prion protein, encoded by PRNP. Since differences in the sequence of PRNP can not explain all variation in TSE susceptibility, there is growing interest in other genes that might have an influence on this susceptibility. One of these genes is SPRN, a gene coding for a protein showing remarkable similarities with the prion protein. Until now, SPRN has not been described in sheep, a highly relevant species in prion matters. RESULTS: In order to characterize the genomic region containing SPRN in sheep, a BAC mini-contig was built, covering approximately 200,000 bp and containing the genes ECHS1, PAOX, MTG1, SPRN, LOC619207, CYP2E1 and at least partially SYCE1. FISH mapping of the two most exterior BAC clones of the contig positioned this contig on Oari22q24. A fragment of 4,544 bp was also sequenced, covering the entire SPRN gene and 1206 bp of the promoter region. In addition, the transcription profile of SPRN in 21 tissues was determined by RT-PCR, showing high levels in cerebrum and cerebellum, and low levels in testis, lymph node, jejunum, ileum, colon and rectum. CONCLUSION: Annotation of a mini-contig including SPRN suggests conserved linkage between Oari22q24 and Hsap10q26. The ovine SPRN sequence, described for the first time, shows a high level of homology with the bovine, and to a lesser extent with the human SPRN sequence. In addition, transcription profiling in sheep reveals main expression of SPRN in brain tissue, as in rat, cow, man and mouse.