APOPTOSIS AND PROLIFERATION IN CANINE PLACENTA

GOMEZ CASTRO G, ZANUZZI C, MERKIS C, BARBEITO C, DIESSLER M.

Congreso; International Federation of Placenta Associations Meeting, Buenos Aires, Argentina September 10 - 13, 2019; 2019

SLIMP-IFPA

Objectives: The aim of this preliminary study was to analyse apoptosis bythe evaluation of active caspase-3 expression and cleaved DNA, and proliferationusing proliferating cell nuclear antigen (PCNA) in late canineplacentae.Methods: Samples of eight canine placentae were collected, formalinfixedand processed by histological routine techniques. Antigen retrievalwas carried out by microwave irradiation. For apoptotic cell detection, arabbit polyclonal anti-active caspase-3 antibody was used, and for cellproliferation a mouse monoclonal anti-PCNA antibody was selected.Reactions were revealed using secondary anti-rabbit or anti-mouseEnVision? detection system and 3,3-diaminobenzidine tetrahydrochloride(DAB). Terminal deoxynucleotidyltransferase UTP nick endlabelling (TUNEL) technique was used to detect DNA fragmentationusing in Situ Cell Death Detection Kit? for immunohistochemistry orimmunofluorescence. Canine small intestine samples were used aspositive controls.Results: Uterine gland epithelium showed apoptotic and proliferating cellswhich were asymmetrically distributed in most of the samples. At junctionalzones we observed several positive apoptotic cell debris. Apoptosisin syncytiotrophoblast was detected with both techniques, while in cytotrophoblastit was only found using TUNEL assay. Besides, some fetal andmaternal blood vessels showed positive apoptotic cells in endothelium andmyocytes.Regarding proliferation cell findings, only moderate labelling of syncytiotrophoblastwas noticed, while most of cytotrophoblast cells resultedpositive. Scant maternal endothelial cells expressed immunoreactivity forPCNA in these late placentae. In addition, proliferation was found inextralaberynthine trophoblast chorionic villi.