Influence of ultrasonic processing on the aggregation of PrP-Sc in the brain extracts of the scrapie-infected hamsters.
- Author:
Xiao-bo ZHOU
1
;
Jian-mei GAO
;
Chen GAO
;
Bao-yun ZHANG
;
Yu-kang YUAN
;
Xiao-ping DONG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Blotting, Western; Brain Chemistry; Cricetinae; Image Processing, Computer-Assisted; PrPSc Proteins; analysis; Scrapie; metabolism; Sonication
- From: Chinese Journal of Experimental and Clinical Virology 2004;18(2):118-121
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUNDTo evaluate the influence ultrasonic processing on the aggregation of PrP(Sc) in the brain extracts prepared from the scrapie-infected hamsters, and to seek for the way to prepare lower molecular PrP(Sc) polymer.
METHODSThe extracts of infected brains were prepared with a lysis solution, and treated with ultrasonics at various conditions during the different phases. The distribution and aggregation state of PrP(Sc) were analyzed by proteinase K treated. Western blot, and afterwards, quantitatively calculated with a commercially supplied software Image Totaltech.
RESULTSThe amount of PrP(Sc) in the supernatant of brain homogenates was 1.29-to 1.58-fold increased with appropriate sonication (15 s for 30 times). At the same conditions of ultrasound, the PrP amount in the supernatant prepared from the scrapie-infected hamster brain was significantly increased, whereas that prepared from healthy animal used as normal control showed little change. Comparative analyses of PrP(Sc) pellets prepared by high-speed centrifugation revealed that about 90% PrP(Sc) released into supernatant after ultrasound processing.
CONCLUSIONAppropriate sonication of homogenate of scrapie-infected brain increases the extracted amount of PrP(Sc), being favorable to laboratory diagnosis. Larger molecular PrP(Sc) aggregates can be crashed by ultrasonic processing, engendering lower molecular PrP(Sc) polymers.