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Advisor(s)
Abstract(s)
Background: Apoptosis detection methods have been proposed for diagnostic purposes, in particular for determining the biological characteristics of tumours and, indirectly, their prognosis.
Implementation of a new technique requires the use of an effective positive control for correct assessment.
Most specialty publications on the subject, do not indicate the positive controls used and do not disclose the obtained results therein.
Objective: The aim of this work was to implement three different methods for the detection of apoptosis, using three canine tissue samples, consisting of breast cancer - carcinoma in situ, thymus and plasmacytoma; and a sample of rabbit tissues - thymus.
Key words: Rabbit, thymus, immunohistochemistry, apoptosis, TUNEL, Bcl-2, Bax.
Methods: Immunohistochemistry was performed, using pro- and antiapoptotic proteins, respectively, Bax and Bcl2 (1: 100; Santa Cruz, Biotechnology) and the TUNEL method (1: 2,5; In situ Cell Death Kit, Roche).
Results
Canine plasmacytoma revealed moderate positive reaction with all methods. Canine carcinoma in situ showed moderate positive reaction with pro-apoptotic antibodies and more intense with anti-apoptotic antibody. Dog’s thymus showed moderate positive reaction in cortical thymocytes with pro-apoptotic antibodies and weak in the medullary thymocytes. With the anti-apoptotic antibody, medullary thymocytes showed weak reaction and negativity in the cortex. Identical results were obtained in rabbit’s thymus, but with stronger intensity.
Conclusions
The rabbit thymus showed the most specific reaction of the tissues studied, by which the authors recommended it as a viable positive control for apoptosis studies.
Description
Keywords
Rabbit thymus immunohistochemistry apoptosis TUNEL Bcl-2 Bax
Pedagogical Context
Citation
Vala H, Santos C, Cruz R, Nóbrega C, Ortiz AL, Mesquita JR, Esteves F, Mega A, (2013). Rabbit thymic tissue – a potential marker for apoptosis techniques. Revista Portuguesa de Ciências Veterinárias. 108 (585-586): 86-87.