absence of detectable fetal microchimerism in nontransgenic goats (Capra aegagrus hircus) bearing transgenic offspring

• Published in: Journal of animal science 2012-02, Vol.90 (2), p.481-488
• Main Authors: Steinkraus, H. B, Rothfuss, H, Jones, J. A, Dissen, E, Shefferly, E, Lewis, R. V
• Format: Article
• Language: English
• Subjects: Animals; Animals, Genetically Modified; Animals, Newborn; Araneae; Autopsy; Birth; blood; Blotting, Western; Bos taurus; Capra; Capra hircus; Casein; Caseins - genetics; cattle; Chimerism; Culling; culling (animals); DNA - chemistry; DNA - genetics; Environmental impact; Female; fetus; Fetuses; Fibroins - analysis; Fibroins - genetics; food chain; Genetic crosses; Genetic engineering; Gestation; goats; Goats - genetics; lactation; Male; Maternal effects; necropsy; parturition; Placenta; Polymerase chain reaction; Polymerase Chain Reaction - veterinary; Pregnancy; progeny; Promoters; protein synthesis; proteins; public health; risk; Ruminantia; Silk; Transgenes; Twins; Western blotting
• ISSN: 0021-8812; 1525-3163
• DOI: 10.2527/jas.2011-4034

Regulations for the disposal of genetically engineered animals are strict due to concern for their inappropriate introduction into the food chain, and of the possible public health and environmental impacts of these organisms. Nontransgenic animals that give birth to transgenic offspring are treated as if they are transgenic due to concern of fetal cells crossing the placental barrier and residing in the mother (fetal-maternal microchimerism). Determining whether or not fetal-fetal or fetal-maternal transfer of DNA or cells occurs during caprine gestation is critical to effectively protect the public without culling animals that pose no risk. Additionally, fetal-maternal transfer, should it exist in the goat, could contraindicate the rebreeding of nontransgenic dams due to the possible transfer of fetal cells from 1 pregnancy to the fetus of subsequent pregnancies. Fetal-maternal transfer in Capra hircus has not been reported in the literature, although it has been reported in another ruminant, Bos taurus. We examined blood from nontransgenic dams that carried transgenic offspring using a PCR method sensitive enough to detect the presence of a spider silk transgene to a 1:100,000 dilution. At this sensitivity, we did not detect the occurrence of fetal-maternal transfer in 5 nontransgenic dams. Likewise, fetal-fetal transfer was not observed from a transgenic to a nontransgenic twin in utero. To test tissue-specific expression of the silk transgene, proteins purified from standard necropsy tissue from a lactating transgenic dam were examined by Western blot analysis. Silk protein expression was only observed in mammary tissue consistent with the tissue specificity of the β-casein promoter used in the transgenic construct. We report evidence collected from a limited caprine breeding pool against transfer of transgenes in utero from fetus to dam and fetus to fetus. In addition, we show evidence that the β-casein promoter in our expression construct is not expressed ectopically as previously suggested. These results suggest that transgene transfer in utero does not occur, but further study is warranted with a larger sample group to confirm these results.