Proteomic Analysis of Chicken Chorioallantoic Membrane (CAM) during Embryonic Development Provides Functional Insight

Abstract

In oviparous animals, the egg contains all resources required for embryonic development. The chorioallantoic membrane (CAM) is a placenta-like structure produced by the embryo for acid-base balance, respiration, and calcium solubilization from the eggshell for bone mineralization. The CAM is a valuable in vivo model in cancer research for development of drug delivery systems and has been used to study tissue grafts, tumor metastasis, toxicology, angiogenesis, and assessment of bacterial invasion. However, the protein constituents involved in different CAM functions are poorly understood. Therefore, we have characterized the CAM proteome at two stages of development (ED12 and ED19) and assessed the contribution of the embryonic blood serum (EBS) proteome to identify CAM-unique proteins. LC/MS/MS-based proteomics allowed the identification of 1470, 1445, and 791 proteins in CAM (ED12), CAM (ED19), and EBS, respectively. In total, 1796 unique proteins were identified. Of these, 175 (ED12), 177 (ED19), and 105 (EBS) were specific to these stages/compartments. This study attributed specific CAM protein constituents to functions such as calcium ion transport, gas exchange, vasculature development, and chemical protection against invading pathogens. Defining the complex nature of the CAM proteome provides a crucial basis to expand its biomedical applications for pharmaceutical and cancer research.

The biological characteristics of chicken embryo mesenchymal stem cells isolated from chorioallantoic membrane

“…RT-PCR showed that Ch-MSCs positively expressed CD29, CD44, CD73, CD90, and CD166, but CD34 and CD45 were not detected (Figure 4b), indicating that Ch-MSCs widely expressed Ch-MSC markers. This is consistent with previous reports (Ahmed et al, 2022;González et al, 2015;Koo et al, 2012).…”

Technical Implications of the Chicken Embryo Chorioallantoic Membrane Assay to Elucidate Neuroblastoma Biology

“…The CAM is a highly vascularized double-membrane structure that is connected to the general circulation of the embryo by two allantoic arteries and one allantoic vein. The CAM is fully differentiated and functional by days 12 and 19, respectively [3,4].…”

“…Proteomic analysis of the CAM on days 12 and 19 has unveiled an extensive array of proteins related to its physiology. These proteins include those involved in calcium ion transport (ATPases, CaBP, and carbonic anhydrases); gas exchange (annexin, hemoglobin, and tubulin); vascularization processes (collagen, integrins, and laminins); and protective molecules against pathogens (cystatins and serpins) [3]. In terms of angiogenesis, the process is predominantly regulated by growth factors with diverse activities including regulatory, proliferative, and inhibitory functions.…”

“…Notably, in the context of CAM, the following regulatory factors have been reportedly involved in this process: VEGF (vascular endothelial growth factor), FGF (fibroblast growth factor), PDGF (platelet-derived growth factor), ANG (angiopoietin), HGF (hepatocyte growth factor), HIF (hypoxia-inducible factor), and endostatin. The chick embryo CAM is a structure commonly used to study blood vessel development and to evaluate molecules with pro-or anti-angiogenic properties [1,3].…”

VEGF Gene Expression and Angiogenesis in the Chorioallantoic Membrane: the Role of Cloprostenol

“…The chorioallantoic membrane (CAM) of a chicken egg is considered a unique experimental model that is applied to trails of blood vessels [1]. Numerous advantages, including simplicity of manipulation, adequacy of naked-eye observations, and inexpensiveness, put this model at the top of experimental approaches were utilized during investigations of blood vessel biology [2].…”