5-BrdU

5-Bromodeoxyuridine (5-BrdU) is an analog of Thy that is incorporated into the DNA of cells undergoing the S-phase and can be detected by monoclonal antibodies or polyclona111.

In vitro: The effect of 5-BrdU on the proliferation was studied in the early phases of dedifferentiation of Nicotiana glauca pith explants grown. It was found to be wholly inhibitory only when given during the first 72 h of culture, this inhibition being reversed by simultaneous addition of either deoxyeytidine or thymidine [1].

In vivo: Young adult offspring mice were stained for 5-BrdU using a monoclonal antibody and the peroxidase-antiperoxidase method. The distribution of immunoreactive nuclei was very similar to the previous [3H]thymidine data. The pattern of labelled nuclei at different embryonic ages followed the spatiotemporal gradients described for cortical and hippocampal neurogenesis. These data indicate that BrdU can be used to map neuronal birthdates. [2].

Clinical trials: Currenlty no clinical data are available.

References:
[1] M.  Durante, C. Geri, V. Nuti-Ronchi, G. Martini, E. Guillé, J. Grisvard, L. Giorgi, R. Parenti, M. Bulatti. Inhibition of Nicotiana glauca pich tissue proliferation through incorporation of 5-BrdU into SNA. Cell Differentiation, Volume 6, Issue 1, June 1977, Pages 53–63
[2] del Rio JA, Soriano E.   Immunocytochemical detection of 5'-bromodeoxyuridine incorporation in the central nervous system of the mouse. Brain Res Dev Brain Res. 1989 Oct 1;49(2):311-7.

[Study of acid soluble proteins of metaphase chromosomes isolated from KB cells before and after 5-BrdU incorporation].[Pubmed:302669]

Ann Genet. 1977 Mar;20(1):25-9.

Metaphase chromosomes and interphase nuclei have been isolated by two different procedures, using either acidic or alkaline media. The morphological integrity of the isolated material has been monitored by electron microscopy. In the acidic procedure the thickness of the chromosome fibers is reduced but the chromosome gross structure is preserved. However, electrophoretic analysis indicates complete disappearance of histones H1 and partial loss of H2A and H2B. No significant modification of acid-soluble proteins isolated from chromosomes or nuclei was observed after 5-BrdU incorporation for 72 hours.

[Compared cytologic study of (in situ and isolated) metaphasic chromosomes from KB cells, after incorporation of 5 bromodeoxyuridine (5 BrdU)].[Pubmed:1085607]

Ann Genet. 1976 Jun;19(2):91.

Substitution of thymidine by 5 BrdU indduces the same morphological and cytological changes in in situ and isolated metaphase chromosomes of KB cells: a dichroic fluorescence and a difference in the affinity of each chromatid for Giemsa stain. However, the precursor does not seem to modify the appearance of G or Q bands in in situ and isolated chromosomes. The fibrilar unit which can be seen with electronic microscopy, shows no difference before and after 5-BrdU incorporation.

Insect sex chromosomes. VIII. Identification of active/inactive X-chromosomes in Gryllotalpa fossor by 5-BrdU/AO fluorescence.[Pubmed:6188623]

Exp Cell Res. 1983 Mar;144(1):228-31.

By employing 5-BrdU/AO fluorescence technique to distinguish active from inactive X-chromosomes, we have, for the first time, provided evidence in support of the validity of this technique for a non-mammalian system Gryllotalpa fossor (Orthoptera). In the female somatic cells of Gryllotalpa, it is only in the active (euchromatic) arm of X-chromosome that the fluorescence is bright, whereas it is dull in the facultative and constitutive heterochromatic arms. In tetraploid spermatogonial cells one of the two X-chromosomes is inactive, suggesting that this phenomenon is probably a random one and that the inactivation process is independent of the imprinting mechanism.

A new method for in vitro detection of bromodeoxyuridine in serum: a proof of concept in a songbird species, the canary.[Pubmed:23691086]

PLoS One. 2013 May 14;8(5):e63692.

Systemic injection of a thymidine analogue such as bromodeoxyuridine (BrdU) in vertebrates is commonly used to detect and study cell production during development, adulthood, and pathology, particularly in studies of adult neurogenesis. Although researchers are applying this technique to multiple species in various physiological conditions, the rate of BrdU clearance from the serum remains unknown in most cases. Changes in this clearance rate as a function of the species, sex or endocrine condition could however profoundly affect the interpretation of the results. We describe a rapid, sensitive, but simple bioassay for post-injection detection and quantification of BrdU in serum. This procedure was shown to be suitable for determining the length of time a thymidine analogue remains in the bloodstream of one avian species and seems applicable to any vertebrate provided sufficiently large blood samples can be collected. This technique was used to demonstrate that, in canaries, BrdU injected at a dose of 100 mg/kg is no longer available for incorporation into DNA between 30 and 60 min post-injection, a delay shorter than anticipated based on the available literature. Preliminary data suggest a similar fast clearance in Japanese quail and mice.

Cell cycle studies based upon quantitative image analysis.[Pubmed:18163464]

Cytometry A. 2008 Apr;73(4):270-8.

When cell cycle studies are performed following cell cycle synchronization, it is possible that critical properties of an actively cycling cell will be overlooked. For this reason past studies have not revealed critical aspects of cell cycle control; such as how a cell determines when to exit the cell cycle, or how rapidly it should cycle. To address these challenging questions we have developed a procedure to quantitate fluorescent stains in a monolayer culture, where nuclear fluorescence and cell cycle history can be assessed with accuracy on a cell by cell basis. The cell cycle position of each cell can be determined by analyzing DNA and BrdU levels. The behavior of cells in a given cell cycle position can then be studied by quantitating up to two other stained markers. When the microinjection of siRNA, neutralizing antibodies, and expression plasmids are coupled with quantitative image analysis, these cell cycle studies can be conducted following alterations in the expression levels of selected cellular targets. With these techniques we have discovered critical aspects of cell cycle control; including how cyclin D1 levels vary through the cell cycle, the molecular mechanisms governing these changes, and the biological implications of changes in cyclin D1 concentration in various cell cycle stages. Our studies with cyclin D1, coupled with similar studies of p27Kip1, form the basis of an entirely new model of cell cycle control proposed here. This model explains how cell cycle progression is terminated, and how the length of the cell cycle is regulated.

5-BrdU is a nucleoside analog that competes with thymidine for incorporation into DNA. 5-BrdU is commonly used in the detection of proliferating cells.

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