Connexin α1 and Cell Proliferation in the Developing Chick Retina

Abstract

During the formation of the eye, high levels of connexin α1 (connexin 43) are expressed within the tissues of the cornea, lens, and neural retina. In order to determine whether connexin α1 plays a role in the regulation of cell proliferation we have used a novel antisense technique to reduce its expression early in development (embryonic days 2–4). Application of Pluronic gel, containing antisense oligodeoxynucleotides (ODNs) to connexin α1, to one eye of early chick embryos results in a rapid and significant reduction of α1 protein which lasts for 24–48 h. Embryos grown for 48 h, after ODN application to one eye, showed a marked reduction in the diameter of the treated, compared to that of the contralateral untreated, eye. Sections cut from the treated eyes showed that the retina was also reduced in size. TUNEL labeling and staining with propidium iodide showed that apoptosis within the retinae of both treated and untreated eyes was rare and thus that the reduction in the area of the retina brought about by antisense ODNs directed at connexin α1 was unlikely to be the result of increased cell death. However, the number of mitotic figures in the ventricular zone of the antisense-treated retinae revealed by propidium iodide staining was significantly reduced (P<0.0001) to 53 ± 3.5% (n=5) of that in the contralateral untreated control eyes. Embryos in which one eye was sham operated, treated with pluronic gel, or treated with sense ODN showed no significant changes in eye size or in the number of mitotic figures within the neural retina. These results point to a role for connexin α1-mediated gap-junctional communication in controlling the early wave of neurogenesis in the chick retina.