Proliferative capacity of embryonic chick notochord: a comparaison between PCNA positivity versus 3H-thymidine incorporation.

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Abstract

The proliferation rate of the embryonic chick notochord is measured during its functional period. Chick embryos of 4 different age groups respectively, 3.5, 4.5, 5.5 and 6.5 days of development, are selected. To measure proliferation activity in the notochords, two different methods are used. 1. Notochords are immunostained for proliferating cell nuclear antigen (PCNA). 2. After in vivo incorporation of 3H-thymidine (3H-TdR), DNA synthesis is visualized by autoradiography. After embedding in paraffin, 5 μm sections are cut and stained with PC10 monoclonal antibody and avidine biotin peroxidase complex (ABC) technique. Corresponding chick embryos of the same age groups are injected in ovo with 3H-thymidine. After fixation 2 μm sections are made and exposed for autoradiography. Results obtained by PCNA and autoradiography labelling techniques for the evaluation of the proliferative capacity are different. Both however show a regression of the proliferative activity. The regression of the number of cells that went through S-phase, as shown by autoradiography, is lower than that of PCNA positive cells. So, a number of notochordal cells stops in G1 phase. Our results moreover, indicate that the proliferating activity of the notochord of chick embryos is higher at young developmental stages than at later stages.

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