Prions: Protein Aggregation and Infectious Diseases - Physiological ...
Prions: Protein Aggregation and Infectious Diseases - Physiological ...
Prions: Protein Aggregation and Infectious Diseases - Physiological ...
You also want an ePaper? Increase the reach of your titles
YUMPU automatically turns print PDFs into web optimized ePapers that Google loves.
1132 ADRIANO AGUZZI AND ANNA MARIA CALELLA<br />
FIG. 9. Role of microglia in prion disease. A: schematic representation for the conditional ablation of microglia in vitro <strong>and</strong> in vivo by using<br />
the cytotoxic prodrug ganciclovir (GCV) in CD11b-HSVTK mice (213). B: histoblot analysis of PrP Sc deposition in POSCA slices after 35 days in<br />
culture shows that PrP Sc accumulates predominantly in the molecular layer. Organotypic brain slices were prepared from 10-day-old tga20 TK (mice<br />
overexpressing PrP <strong>and</strong> carrying the CD11b-HSVTK transgene) <strong>and</strong> were inoculated with 100 g RML (R) or uninfected homogenate (Ø). PrP Sc was<br />
detected with anti-PrP antibody (POM1). C: accumulation of PrP Sc in slices was also shown by Western blot; tga20 TK <strong>and</strong> prnp o/o slices were<br />
cultured for 35 days, optionally digested with PK (g/ml), <strong>and</strong> probed with antibody POM1. D: microglia depletion in organotypic slices increases<br />
the amount of PK-resistant PrP. Organotypic slices from mice overexpressing PrP with the CD11b-HSVTK transgene (tga20 TK ) or without<br />
(tga20 TK ) were treated with GCV <strong>and</strong> inoculated with RML. Samples were PK-digested <strong>and</strong> PrP detected with POM1. E: microglia depletion in<br />
organotypic slices increases the infectivity as evaluated by SCEPA of homogenates of tga20 TK or prnp o/o/TK slices treated with GCV. Three<br />
independent biological replicas of tga20 TK <strong>and</strong> single replicas of prnp o/o/TK slices or RML were analyzed in 10-fold dilution steps using 6–12<br />
replica N2a-containing wells per dilution. Data are indicated as the number of infectious tissue culture units per gram of slice culture protein <strong>and</strong><br />
are the averages of biological replicas SD. Slices from the same animal (pairs of GCV <strong>and</strong> GCV samples) are represented by the same color.<br />
Sc, positive-control homogenate from brain of a mouse with scrapie; Inoc, inoculum. Left lane on all blots, molecular weight marker spiked with<br />
recombinant PrP C , yielding a PrP signal at 23 kDa with a cleavage product at 15 kDa. [B–E are adapted from Falsig et al. (149).]<br />
Physiol Rev VOL 89 OCTOBER 2009 www.prv.org