Abstract

Simple SummaryThe present study compared the effectivity of two cell proliferation markers, BrdU and EdU, in the brain neurogenic zone of the songbird zebra finch. It shows their saturation doses, that BrdU labels more cells than the equimolar dose of EdU, and that both markers can be reliably detected in the same brain.There are two most heavily used markers of cell proliferation, thymidine analogues 5-bromo-2′-deoxyuridine (BrdU) and 5-ethynyl-2′-deoxyuridine (EdU) that are incorporated into the DNA during its synthesis. In neurosciences, they are often used consecutively in the same animal to detect neuronal populations arising at multiple time points, their migration and incorporation. The effectivity of these markers, however, is not well established. Here, we studied the effectivity of equimolar doses of BrdU and EdU to label new cells and looked for the dose that will label the highest number of proliferating cells in the neurogenic ventricular zone (VZ) of adult songbirds. We found that, in male zebra finches (Taeniopygia guttata), the equimolar doses of BrdU and EdU did not label the same number of cells, with BrdU being more effective than EdU. Similarly, in liver, BrdU was more effective. The saturation of the detected brain cells occurred at 50 mg/kg BrdU and above 41 mg/kg EdU. Higher dose of 225 mg/kg BrdU or the equimolar dose of EdU did not result in any further significant increases. These results show that both markers are reliable for the detection of proliferating cells in birds, but the numbers obtained with BrdU and EdU should not be compared.

Highlights

  • The phenomenon of adult neurogenesis occurs in a wide variety of species including mammals and birds

  • Paired t-tests were used to compare the cell counts obtained by two anti-bromo-20 -deoxyruidine (BrdU) antibodies, BrdU and ethynyl-2 -deoxyuridine (EdU) as well as to compare two concentrations (1:1000 and 1:200) of the MoBU-1 antibody and EdU with and without DNA denaturation

  • We found that the revealed that there is no significant difference in fluorescent labeling between these two antibodies increasing doses of BrdU and EdU led to increases in the numbers of labeled cells in the ventricular zone (VZ)

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Summary

Introduction

The phenomenon of adult neurogenesis occurs in a wide variety of species including mammals and birds. The newborn cells arise from progenitor cells in specific brain regions, migrate to their ultimate destinations and differentiate. The new neurons in mammals are born in the subventricular zone (SVZ) of the lateral ventricles or in the subgranular zone of the dentate gyrus in hippocampus [1]. The proliferating cells in birds are located in the walls of the lateral ventricle in the ventricular zone (VZ) [2]. The accurate and effective labeling of the newborn cells is critical for the study of neurogenesis. There are several other methods for the detection of DNA synthesis [3], the marker of choice in recent years is the thymidine analogue 5-bromo-20 -deoxyruidine (BrdU) that is incorporated

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