Title: 2D-DIGE analysis of differential protein expression in the chick embryo chorioallantoic membrane as a consequence of increased carbon dioxide in the incubator during the first 10 days.
Abstract: Previous experiments in which the effect of a gradual increase of the carbon dioxide (CO 2) concentration (up to 1.5% during the first 10 days of incubation) on the development of the chick embryo was studied, revealed a faster embryonic growth, higher pCO 2 and lower pO 2 in the air cell till day 16, a faster hatch with a narrower spread and different thyroid hormone and corticosterone plasma concentrations for the chick embryos of the non-ventilated (increased CO 2) incubation. In addition, some persistent effects during the post-hatch period were recorded, such as a faster growth for the non-ventilated chicks. During embryonic development, the chorioallantoic membrane (CAM) plays an important role in the respiratory interchange of oxygen and carbon dioxide and in the uptake of Ca 2+ from the shell. The CAM is an extraembryonic tissue formed by fusion of the chorion and the allantois during early avian development. While the CAM is in close contact with the atmosphere, it might be affected by the increased CO 2-concentration in the incubator. To reveal the underlying mechanisms that might cause the increased growth speed, two dimensional differential gel electrophoresis (2D-DIGE) was used to analyse possible changes in protein expression patterns of the CAM of 12 day old chick embryos, the time point at which the CAM is completely developed and fully functional. Eight tissue samples per group were used for analysis. Isoelectric focusing of the protein extracts was carried out using IPG strips with a non linear pH range of 3 to 10 followed by SDS-page. In the CAM, 23 significantly different protein spots were detected, from which 15 were identified unambiguously. Remarkable was, that all differential proteins had a higher expression in control chicks. All proteins were thus down regulated under the influence of higher ambient CO 2. Identification of the differential spots with mass spectrometry revealed that these proteins are involved in different physiological mechanisms including oxidative stress, cytoskeletal organisation, energy metabolism, Ca 2+ binding. This experiment shows that differential CO2 concentrations during the first half of incubation have an impact on several physiological pathways in the CAM. It remains however to be elucidated how these changes are linked with the observed effects on the embryo itself.
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
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