NR AWJI
AU Lyahyai,J.; Serrano,C.; Marcos-Carcavilla,A.; Bolea,R.; Badiola,J.J.; Zaragoza,P.; Martin-Burriel,I.
TI Differential expression of BCL-2 family genes in central nervous system of sheep with natural scrapie
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PA-33
PT Konferenz-Poster
AB Neuronal loss is a salient feature of prion disease; however, its causes and mechanisms are unclear. Some experimental studies show that neuronal death could occur through an apoptotic process. In the present study this process was analysed in the Central Nervous System (CNS) of seven naturally scrapie infected sheep and five breed (Rasa Aragonesa), sex (female), age (3-5 years) and PRNP genotype (ARQ/ARQ) matched controls. In situ end labelling (TUNEL) and immunodetection of the activated form of caspase-3 were used to identify apoptotic cells in different brain regions from scrapie and control sheep. Both methods revealed a reduced number of stained glial cells and a very low amount of positive neurons with apoptotic appearance. These results could be expected as only a few neurons are supposed to die at a given time point in a chronic disease like natural scrapie. What is observable using these techniques may represent a minute proportion of all apoptotic events going on in TSE affected brains. The analysis of early apoptosis related markers could facilitate both, the identification of this process and the determination of the molecular pathways involved. We have investigated the existence of differences in transcript levels of genes involved in the mitochondria pathway (BAX, BCL-2, BCL-XL, BCL-XS, BCL-W, BAD, BAK, MCL-1, CYT-C, HSP-27 and APAF-1). When sheep sequences were known, specific primers were designed to amplify cDNA fragments. In most cases, heterologous primers were used to identify sheep genes and, after sequencing, to design specific primers. Gene expression was quantified using Real Time RT-PCR. PCR reactions were performed using the TaqMan and SYBR-Green assays. GAPDH, ACTB and 18S rRNA, three of the most common housekeeping genes, were used as internal controls against which all samples were normalized. Expression results indicate that genes belonging to the mitochondrial pathway are involved in the neuropathology observed in scrapie.
AD J. Lyahyai, C. Serrano, P. Zaragoza, I. Martín-Burriel: LAGENBIO. Universidad de Zaragoza, Spain; A. Marcos-Carcavilla: INIA. Madrid, Spain; R. Bolea, J.J. Badiola: CNREET. Universidad de Zaragoza, Spain. E-mail: jaberfly2@hotmail.com
SP englisch
PO Italien