Cytotoxic Effect of 5-Bromo-2-Deoxyuridine on Olfactory Epithelium

In vertebrates the most notable feature of the olfactory epithelium (OE) is its ability to continually replace cell types, including mature sensory neurons. In amphibians, the neuronal turnover is observed in adult animals and during larval development also. Today, one of the most used techniques for studies of cell proliferation and differentiation is the 5-bromo-2-deoxyuridine (BrdU) uptake, a synthetic analogue of thymidine that is incorporated into cells during DNA replication. Despite this, and because of its toxicity, some authors have questioned its use for the above mentioned purposes. This work analyses BrdU incorporation technique for cell proliferation and differentiation studies within the OE of amphibian larvae. The methodology used has been validated in Xenopus laevis larvae for studies related to inner ear formation. However, we observe a large number of adverse effects on cellular components of the OE, such as an increase in apoptosis, alterations in the expression pattern of different cell markers, as well as defects in the foraging behavior in animals that were exposed to BrdU. We conclude that even though BrdU can be successfully used as a proliferation cellular marker in the OE, It should be very careful when BrdU is used for studying cell lineage.

[1]  D. Paz,et al.  A putative functional vomeronasal system in anuran tadpoles , 2012, Journal of anatomy.

[2]  S. Shioda,et al.  Abnormal brain function of the rat neonate in a prenatal 5-bromo-2′-deoxyuridine (BrdU)-induced developmental disorder model , 2012, International Journal of Developmental Neuroscience.

[3]  F. d’Adda di Fagagna,et al.  Neural stem cells exposed to BrdU lose their global DNA methylation and undergo astrocytic differentiation , 2012, Nucleic acids research.

[4]  P. Rakic,et al.  Different Effects of Bromodeoxyuridine and [3H]Thymidine Incorporation into DNA on Cell Proliferation, Position, and Fate , 2011, The Journal of Neuroscience.

[5]  P. Coumailleau,et al.  Proliferation, migration and differentiation in juvenile and adult Xenopus laevis brains , 2011, Brain Research.

[6]  K. Kaga,et al.  Age‐related changes in cell dynamics of the postnatal mouse olfactory neuroepithelium: Cell proliferation, neuronal differentiation, and cell death , 2010, The Journal of comparative neurology.

[7]  D. Paz,et al.  Disruptive Effect of Epigallocatechin-3-Gallate on the Proliferation/Apoptosis Balance in the Olfactory Epithelium~!2009-11-15~!2010-02-05~!2010-03-05~! , 2010 .

[8]  D. Paz,et al.  Amphibian larvae and zinc sulphate: a suitable model to study the role of brain-derived neurotrophic factor (BDNF) in the neuronal turnover of the olfactory epithelium , 2009, Cell and Tissue Research.

[9]  R. Beyer,et al.  Transcriptional biomarkers and mechanisms of copper-induced olfactory injury in zebrafish. , 2008, Environmental science & technology.

[10]  D. Schild,et al.  Nucleotide‐induced Ca2+ signaling in sustentacular supporting cells of the olfactory epithelium , 2008, Glia.

[11]  P. Ross,et al.  Salmon olfaction is impaired by an environmentally realistic pesticide mixture. , 2008, Environmental science & technology.

[12]  J. Arellano,et al.  Everything that glitters isn't gold: a critical review of postnatal neural precursor analyses. , 2007, Cell stem cell.

[13]  W. Wilczynski,et al.  Regional distribution and migration of proliferating cell populations in the adult brain of Hyla cinerea (Anura, Amphibia) , 2007, Brain Research.

[14]  P. Ross,et al.  Linuron and carbaryl differentially impair baseline amino acid and bile salt olfactory responses in three salmonids. , 2007, Toxicology.

[15]  P. Taupin BrdU immunohistochemistry for studying adult neurogenesis: Paradigms, pitfalls, limitations, and validation , 2007, Brain Research Reviews.

[16]  D. Paz,et al.  Immunohistochemical localization of vascular endothelial growth factor and its receptor Flk-1 in the amphibian developing principal and accessory olfactory system , 2006, Anatomy and Embryology.

[17]  Clive N Svendsen,et al.  5‐Bromo‐2′‐deoxyuridine is selectively toxic to neuronal precursors in vitro , 2005, The European journal of neuroscience.

[18]  E. Mugnaini,et al.  Time of origin of unipolar brush cells in the rat cerebellum as observed by prenatal bromodeoxyuridine labeling , 2004, Neuroscience.

[19]  C. Gross,et al.  Neurogenesis in Adult Mammals: Some Progress and Problems , 2002, The Journal of Neuroscience.

[20]  N. L. Hayes,et al.  Stem Cells: The Promises and Pitfalls , 2001, Neuropsychopharmacology.

[21]  V H Casco,et al.  Expression of isoforms of the neural cell adhesion molecule (NCAM) and polysialic acid during the development of the Bufo arenarum olfactory system. , 1995, The International journal of developmental biology.

[22]  S. Youngentob,et al.  Reconstitution of the rat olfactory epithelium after methyl bromide‐induced lesion , 1995, The Journal of comparative neurology.

[23]  J. Rafols,et al.  Histological and histochemical studies of the secretory components of the salamander olfactory mucosa: Effects of isoproterenol and olfactory nerve section , 1984, The Anatomical record.

[24]  J. Rafols,et al.  Morphological relations between the receptor neurons, sustentacular cells and Schwann cells in the olfactory mucosa of the salamander , 1983, The Anatomical record.

[25]  R. Davidson,et al.  Total substitution of bromodeoxyuridine for thymidine in the DNA of a bromodeoxyuridine-dependent cell line. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[26]  K. Gosner,et al.  A simplified table for staging anuran embryos and larvae with notes on identification , 1960 .

[27]  E. Serrano,et al.  Cell proliferation during the early compartmentalization of the Xenopus laevis inner ear. , 2007, The International journal of developmental biology.

[28]  P. Rakic Neurogenesis in adult primates. , 2002, Progress in brain research.