Dual BrdU-PCNA immunodetection of proliferative cells in dental and orofacial tissues of teleosts


Author’s address: Sam Vandenplas, Biology Department, Ledeganckstraat 35 B-9000 Gent, Belgium.
E-mail: sam.vandenplas@ugent.be


We have developed a protocol for simultaneous detection of BrdU and PCNA on paraffin sections of teleost tissues and have tested it successfully on dental and orofacial tissues of Atlantic salmon (Salmo salar) and zebrafish (Danio rerio). By combining decalcification of the tissue, appropriate antigen retrieval steps, and high quality tissue sections, this protocol allows to study the instantaneous pattern of proliferating cells, as well as cell tracing in large specimens. The technique is therefore highly suitable to study skeletal and dental tissues, tissues that are traditionally not easily accessible to molecular and cellular techniques. Dual BrdU-PCNA immunostaining may serve as a useful tool in experimental studies on fish developmental or regenerative processes such as tissue turnover in orofacial epithelia, continuous tooth replacement, and in the identification of putative stem cells for which the cycling state of BrdU label retaining cells needs to be determined.