Detection of CWD prions in naturally infected white-tailed deer fetuses and gestational tissues by PMCA

Abstract

Chronic wasting disease (CWD) is a prevalent prion disease affecting cervids. CWD is thought to be transmitted through direct animal contact or by indirect exposure to contaminated environmental fomites. Other mechanisms of propagation such as vertical and maternal transmissions have also been suggested using naturally and experimentally infected animals. Here, we describe the detection of CWD prions in naturally-infected, farmed white-tailed deer (WTD) fetal tissues using the Protein Misfolding Cyclic Amplification (PMCA) technique. Prion seeding activity was identified in a variety of gestational and fetal tissues. Future studies should demonstrate if prions present in fetuses are at sufficient quantities to cause CWD after birth. This data confirms previous findings in other animal species and furthers vertical transmission as a relevant mechanism of CWD dissemination.

Figure 1

PrP^C and PrP^Sc screening in white-tailed deer fetal tissues using western blot and PMCA. Western blot analysis of representative fetal tissue samples prior (A) and after (B) PK treatment. (C) Results from the same representative samples depicted in (A) and (B) after PMCA analysis. Numbers at the right of each panel represent molecular weight markers (in KDa). “PrP^C” denotes brain extracts from Tg1536 mice not treated with PK and used as additional molecular weight markers. The solid line within panel 1C denote images taken from different membranes.

Figure 2

Identification of PrP^Sc in white-tailed deer gestational tissues and fluids by western blot and PMCA. Western blot analysis of representative gestational tissue samples prior (A) and after (B) PK treatment. (C) PMCA results from the same representative samples depicted in (A) and (B) after three PMCA rounds. Numbers at the right of each panel represent molecular weight markers (in KDa). “PrP^C” denotes brain extracts from Tg1536 mice not treated with PK and used as additional molecular weight markers.