Publication Database

Cry1Ab treatment has no effects on Viability of Cultured Porcine Intestinal Cells, but Triggers Hsp70 Expression (2013)

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Bondzio, Angelika (WE 3)

Lodemann, Ulrike (WE 2)

Weise, Christoph

Einspanier, Ralf (WE 3)

Forschungsprojekt

SFB 852-TP INF: Zentrales Technik- und Bioinformatikprojekt

Quelle

PLOS ONE

Bandzählung: 8

Heftzählung: 7

Seiten: e67079

ISSN: 1932-6203

Sprache

Englisch

Verweise

URL (Volltext): http://edocs.fu-berlin.de/docs/receive/FUDOCS_document_000000019452

DOI: 10.1371/journal.pone.0067079

Pubmed: 23861753

Kontakt

Institut für Veterinär-Biochemie

Oertzenweg 19 b
14163 Berlin
+49 30 838 62225
biochemie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

In vitro testing can contribute to reduce the risk that the use of genetically modified (GM) crops and their proteins show unintended toxic effects. Here we introduce a porcine intestinal cell culture (IPEC-J2) as appropriate in vitro model and tested the possible toxic potential of Cry1Ab protein, commonly expressed in GM-maize. For comprehensive risk assessment we used WST-1 conversion and ATP content as metabolic markers for proliferation, lactate dehydrogenase release as indicator for cells with compromised membrane and transepithelial electrical resistance as parameter indicating membrane barrier function. The results were compared to the effects of valinomycin, a potassium ionophore, known to induce cytotoxic effects in most mammalian cell types. Whereas no toxicity was observed after Cry1Ab treatment, valinomycin induced a decrease in IPEC-J2 viability. This was confirmed by dynamic monitoring of cellular responses. Additionally, two dimensional differential in-gel electrophoresis was performed. Only three proteins were differentially expressed. The functions of these proteins were associated with responses to stress. The up-regulation of heat shock protein Hsp70 was verified by Western blotting as well as by enzyme-linked immunosorbent assay and may be related to a protective function. These findings suggest that the combination of in vitro testing and proteomic analysis may serve as a promising tool for mechanism based safety assessment.