Brdu birthdating protocol
One advantage of Brd U is that it can be quickly detected by immunocytochemistry.Moreover, it can be used in combination with immunoperoxidase labeling for phenotypic markers, which can then be detected by bright-field, fluorescence, and confocal microscopy to determine colocalization (e.g., ).Because [H]-thymidine and Brd U label cells for only about 2 h after injection  and perhaps for an even shorter time in birds , the time of cell division can be determined.Both markers have advantages and drawbacks (reviewed in ) and hence are preferable for some applications and limited for others.Department of Natural and Life Sciences, The Open University of Israel, 108 Ravotsky Street, P. Box 808, 43107 Ra’anana, Israel Received 10 June 2014; Accepted 26 October 2014Academic Editor: Giovanni Scapagnini Copyright © 2015 Anat Cattan et al.This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.A disadvantage of Brd U is that its molecular structure inherently differs from the natural structure of thymidine, causing steric hindrance (Figure 1).Consequently, Brd U is a toxic mutagenic substance and can cause abnormalities in DNA transcription and protein translation, which could lead to mutation and cell toxicity, compromising the overall health and behavior of the organisms, both in fetal and in postdevelopmental stages [14–16].
Whereas all these studies were performed on mammals, in birds such a thorough examination has not been conducted.In one of our previous studies, we encountered surprising results in a study with zebra finches (Taeniopygia guttata).To provide evidence for neuronal replacement, we used two cell birth markers in the same birds: Brd U, to label neurons born 1 or 3 months prior to social manipulation , and [ neurons to appear also in the recent study, but this was not the case.Accordingly, we examined whether Brd U might alter neurogenesis in the adult avian brain.We compared recruitment of [ neuronal nucleus and their distribution were similar across groups. The results indicate that the commonly used dosage of Brd U in birds has no long-term effects on subsequent cell divisions and neuronal recruitment.
In addition, Brd U triggers cell death and the formation of teratomas, alters DNA stability, lengthens cell cycle, and has mitogenic, transcriptional, and translational effects on the cells that incorporate it (reviewed in ).