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Title: Expression of the ephrin receptor B2 in the embryonic chicken bursa of Fabricius

Author
item WEATHERS, B - Mississippi State University
item Branton, Scott
item JACOB, R - Mississippi State University
item TAYLOR, JR, R - West Virginia University
item PEEBLES, E - Mississippi State University
item PHARR, G - Mississippi State University

Submitted to: International Journal of Poultry Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/25/2015
Publication Date: 11/30/2015
Citation: Weathers, B., Branton, S.L., Jacob, R., Taylor, Jr, R.L., Peebles, E.D., Pharr, G.T. 2015. Expression of the ephrin receptor B2 in the embryonic chicken bursa of Fabricius. International Journal of Poultry Science. 14(9):485-490.

Interpretive Summary: Investigations with poultry lighting studies have recently included assessment of immune function. One parameter of immune function is associated with bursal (bursa of Fabricius [[BOF]) weight as a function of the total body weight of the chicken. The BOF is a specific anatomical site wherein chicken B-cells develop. However, to understand the BOF microenvironment guiding B-cell development, previous studies identified ephrin (Eph) receptor B2 (EphB2) gene transcripts in the embryonic bursa. We hypothesize that the EphB2 receptors and their ligands regulate interactions between developing B-cells and epithelial cells in the embryonic BOF follicle. To address this hypothesis, additional basic studies were conducted to determine if EphB2 is expressed in the embryonic BOF. Therefore, EphB2 gene expression was examined with reverse transcriptase-PCR and western blotting in the BOF on embryonic days (ED) 15 and 18. The RT-PCR experiments were conducted with BOF cDNA amplified transcripts from the EphB2 gene, which was confirmed by nucleotide sequencing. The anti-EphB2 antibody recognized a protein of the expected 120 kDa molecular mass and gave an acceptable signal to noise ratio on whole tissue protein western blots from the ED15 and ED18 bursa. This information can potentially be used to design experiments for determining the cell type(s) expressing the EphB2 protein in the embryonic bursa. Then, practical poultry lighting studies can be conducted and the expression of EphB2 protein can be quantitated thereby providing a measure for comparing one lighting regime against a different lighting regime or light source.

Technical Abstract: Chicken B-cells develop in a specific organ, the bursa of Fabricius. To understand the bursal microenvironment guiding B-cell development, previous studies identified ephrin (Eph) receptor B2 (EphB2) gene transcripts in the embryonic bursa. We hypothesize that the EphB2 receptors and their ligands regulate interactions between developing B-cells and epithelial cells in the embryonic bursal follicle. To address this hypothesis, additional basic studies were conducted to determine if EphB2 is expressed in the embryonic bursa. Therefore, EphB2 gene expression was examined with reverse transcriptase-PCR and western blotting in the bursa on embryonic days (ED) 15 and 18. The RT-PCR experiments with bursal cDNA amplified transcripts from the EphB2 gene, which was confirmed by nucleotide sequencing. The anti-EphB2 antibody recognized a protein of the expected 120 kDa molecular mass and gave an acceptable signal to noise ratio on whole tissue protein western blots from the ED15 and ED18 bursa. This information can be used to design experiments for determining the cell type(s) expressing the EphB2 protein in the embryonic bursa.