Bromodeoxyuridine: Difference between revisions

{{short description|Compound commonly used to detect proliferating cells}}

{{Technical|date=February 2021}}

| Verifiedfields = changed

| Watchedfields = changed

| verifiedrevid = 401935585

| ImageFile=Bromodeoxyuridine.svg

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| IUPACName=5-Bromo-2′-deoxyuridine

| SystematicName=5-Bromo-1-[(2''R'',4''S'',5''R'')-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidine-2,4(1''H'',3''H'')-dione

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|Section1={{Chembox Identifiers

| ChemSpiderID_Ref = {{chemspidercite|correct|chemspider}}

| ChemSpiderID = 5294121

| ChEMBL_Ref = {{ebicite|changed|EBI}}

| ChEMBL = 222280

| PubChem = 6035

| ChemSpiderID2_Ref = {{chemspidercite|changed|chemspider}}

| ChemSpiderID2 = 5813

| SMILES = c1c(c(=O)[nH]c(=O)n1[C@H]2C[C@@H]([C@H](O2)CO)O)Br

| InChI = 1/C9H11BrN2O5/c10-4-2-12(9(16)11-8(4)15)7-1-5(14)6(3-13)17-7/h2,5-7,13-14H,1,3H2,(H,11,15,16)/t5-,6+,7+/m0/s1

| InChIKey = WOVKYSAHUYNSMH-RRKCRQDMBM

| StdInChI_Ref = {{stdinchicite|changed|chemspider}}

| StdInChI = 1S/C9H11BrN2O5/c10-4-2-12(9(16)11-8(4)15)7-1-5(14)6(3-13)17-7/h2,5-7,13-14H,1,3H2,(H,11,15,16)/t5-,6+,7+/m0/s1

| StdInChIKey_Ref = {{stdinchicite|changed|chemspider}}

| StdInChIKey = WOVKYSAHUYNSMH-RRKCRQDMSA-N

| CASNo_Ref = {{cascite|correct|CAS}}

| MeSHName=Bromodeoxyuridine

|Section2={{Chembox Properties

| C=9 | H=11 | Br=1 | N=2 | O=5

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|Section3={{Chembox Hazards

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'''Bromodeoxyuridine''' ('''5-bromo-2'-deoxyuridine''', '''BrdU''', '''BUdR''', '''BrdUrd''', '''broxuridine''') is a synthetic [[nucleoside analogue]] with a chemical structure similar to [[thymidine]]. BrdU is commonly used to study [[cell proliferation]] in living tissues<ref>{{cite journal |doi=10.1007/s00441-011-1213-7 |title=The dark side of BrdU in neural stem cell biology: Detrimental effects on cell cycle, differentiation and survival |year=2011 |last1=Lehner |first1=Bernadette |last2=Sandner |first2=Beatrice |last3=Marschallinger |first3=Julia |last4=Lehner |first4=Christine |last5=Furtner |first5=Tanja |last6=Couillard-Despres |first6=Sebastien |last7=Rivera |first7=Francisco J. |last8=Brockhoff |first8=Gero |last9=Bauer |first9=Hans-Christian |journal=Cell and Tissue Research |pmid=21837406 |volume=345 |issue=3 |pages=313–28 |last10=Weidner |first10=Norbert |last11=Aigner |first11=Ludwig|s2cid=756261 }}</ref> and has been studied as a [[radiosensitizer]]<ref name="Cancer Research">{{cite journal |vauthors=Russo A, Gianni L, Kinsella TJ, Klecker RW, Jenkins J, Rowland J, Glatstein E, Mitchell JB, Collins J, Myers C |title=Pharmacological evaluation of intravenous delivery of 5-bromodeoxyuridine to patients with brain tumors |journal=Cancer Res. |volume=44 |issue=4 |pages=1702–5 |year=1984 |pmid=6704976 }}</ref> and [[medical diagnosis|diagnostic tool]] in people with [[cancer]].<ref>{{cite journal |last1=Dolbeare |first1=F |title=Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part I: Historical perspectives, histochemical methods and cell kinetics |journal=The Histochemical Journal |date=May 1995 |volume=27 |issue=5 |pages=339–69 |doi=10.1007/BF02389022 |pmid=7657555|s2cid=21785471 }}</ref>

During the [[S phase]] of the [[cell cycle]] (when [[DNA replication]] occurs), BrdU can be incorporated in place of thymidine in newly synthesized [[DNA]] molecules of dividing cells.<ref name="Ki-67 V. BrdU">{{cite journal|last=Kee| first=N |author2=S Sivalingam |author3=R Boonstra |author4=J.M Wojtowicz|title=The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis|journal=Journal of Neuroscience Methods|date=March 2002|volume=115|issue=1| pages=97–105|doi=10.1016/S0165-0270(02)00007-9|pmid=11897369|hdl=1807/357 |s2cid=17572667 |hdl-access=free}}</ref> Cells that have recently performed DNA replication or [[DNA repair]] can be detected with [[antibody|antibodies]] specific for BrdU using techniques such as [[immunohistochemistry]] or [[immunofluorescence]].<ref name=pmid21785232>{{cite journal |pages=433–40 |doi=10.1269/jrr.10097 |title=Visualization of Heavy Ion Tracks by Labeling 3'-OH Termini of Induced DNA Strand Breaks |year=2011 |last1=Konishi |first1=Teruaki |last2=Takeyasu |first2=Akihiro |last3=Natsume |first3=Toshiyuki |last4=Furusawa |first4=Yoshiya |last5=Hieda |first5=Kotaro |journal=Journal of Radiation Research |volume=52 |issue=4 |pmid=21785232|bibcode=2011JRadR..52..433K |doi-access= }}</ref> BrdU-labelled cells in humans can be detected up to two years after BrdU infusion.<ref name="Nature Neurogenesis">{{cite journal|last=Eriksson|first= Peter |author2=Ekaterina Perfilieva |author3=Thomas Björk-Eriksson |author4=Ann-Marie Alborn |author5=Claes Nordborg |author6=Daniel A. Peterson |author7=Fred H. Gage|title=Neurogenesis in the adult human hippocampus|journal=Nature Medicine|year=1998|volume=4|series=1313-1317|pages=1313–1317|doi=10.1038/3305|pmid=9809557|issue=11|doi-access=free}}</ref>

Because BrdU can replace [[thymidine]] during DNA replication, it can cause [[mutation]]s, and its use is therefore potentially a health hazard.{{cn|date=March 2021}} However, because it is neither [[radioactive]] nor [[myelotoxic]] at labeling concentrations, it is widely preferred for ''in vivo'' studies of [[cancer]] cell proliferation.<ref name="Fujimaki Cancer">{{cite journal|last=Fujimaki |first=Takamitsu |author2=Masao Matsutani |author3=Osamu Nakamura |author4=Akio Asai |author5=Nobuaki Funada |author6=Morio Koike |author7=Hiromu Segawa |author8=Kouichi Aritake |author9=Takanori Fukushima |author10=Shuntaro Houjo |author11=Akira Tamura |author12=Keiji Sano |title=Correlation Between Bromodeoxyuridine- Labeling Indices and Patient Prognosis in Cerebral Astrocytic Tumors of Adults|journal=Cancer|date=1991|volume=67|issue=6|pages=1629–1634|doi=10.1002/1097-0142(19910315)67:6<1629::AID-CNCR2820670626>3.0.CO;2-E|pmid=2001552|doi-access=free }}</ref><ref name="Cytometry Myelotoxic">{{cite journal|last=Hoshino|first=Takao|author2=Tadashi Nagashima|author3=Judith Murovic|author4=Ellen M. Levin|author5=Victor A. Levin|author6=Stephen M. Rupp|title=Cell Kinetic Studies of In Situ Human Brain Tumors With Bromodeoxyuridine|journal=Cytometry|year=1985|volume=6|issue=6|pages=627–632|doi=10.1002/cyto.990060619|pmid=2998714|doi-access=free}}</ref> However, at [[radiosensitizing]] concentrations, BrdU becomes myelosuppressive, thus limiting its use for radiosensitizing.<ref name="Cancer Research" />

BrdU differs from [[thymidine]] in that BrdU substitutes a [[bromine]] atom for [[thymidine|thymidine's]] CH₃ group. The Br substitution can be used in X-ray diffraction experiments in crystals containing either DNA or RNA. The Br atom acts as an anomalous scatterer and its larger size will affect the crystal's X-ray diffraction enough to detect [[Isomorphism (crystallography)|isomorphous]] differences as well.<ref>{{cite journal |pages=24–34 |doi=10.1107/S0909049595013288 |title=MAD Phasing Strategies Explored with a Brominated Oligonucleotide Crystal at 1.65Å Resolution |year=1996 |last1=Peterson |first1=M. R. |last2=Harrop |first2=S. J. |last3=McSweeney |first3=S. M. |last4=Leonard |first4=G. A. |last5=Thompson |first5=A. W. |last6=Hunter |first6=W. N. |last7=Helliwell |first7=J. R. |author-link7=John R. Helliwell|journal=Journal of Synchrotron Radiation |volume=3 |pmid=16702655 |issue=Pt 1|doi-access=free |bibcode=1996JSynR...3...24P }}</ref><ref>{{cite journal |pages=374–80 |doi=10.1107/S0907444909051609 |pmc=2852301 |title=The magic triangle goes MAD: Experimental phasing with a bromine derivative |year=2010 |last1=Beck |first1=Tobias |last2=Gruene |first2=Tim |last3=Sheldrick |first3=George M. |journal=Acta Crystallographica Section D |volume=66 |issue=4 |pmid=20382990|bibcode=2010AcCrD..66..374B }}</ref>

Bromodeoxyuridine releases gene silencing caused by DNA methylation.<ref>{{cite journal |vauthors=Weiss RA |title=On the concept and elucidation of endogenous retroviruses |journal=Philos. Trans. R. Soc. Lond. B Biol. Sci. |volume=368 |issue=1626 |pages=20120494 |year=2013 |pmid=23938748 |pmc=3758183 |doi=10.1098/rstb.2012.0494 }}</ref>

BrdU can also be used to identify microorganisms that respond to specific carbon substrates in aquatic<ref>{{cite journal|last1=Tada|first1=Yuya|last2=Grossart|first2=Hans-Peter|title=Community shifts of actively growing lake bacteria after N-acetyl-glucosamine addition: improving the BrdU-FACS method|journal=The ISME Journal|volume=8|issue=2|year=2013|pages=441–454|issn=1751-7362|doi=10.1038/ismej.2013.148|pmid=23985742|pmc=3906810}}</ref> and soil<ref>{{cite journal |vauthors=Borneman J |title=Culture-independent identification of microorganisms that respond to specified stimuli |journal=Appl. Environ. Microbiol. |volume=65 |issue=8 |pages=3398–400 |year=1999 |pmid=10427025 |pmc=91510 |doi= 10.1128/AEM.65.8.3398-3400.1999|bibcode=1999ApEnM..65.3398B }}</ref> environments. A carbon substrate added to the incubations of environmental samples will cause the growth of microorganisms that can utilize that substrate. These microorganisms will then incorporate BrdU into their DNA as they grow. Community DNA can then be isolated and BrdU-labeled DNA purified using an immunocapture technique.<ref>{{cite journal|last=Urbach|first=Ena|author2=Kevin L. Vergin|author3=Stephen J. Giovannoni|title=Immunochemical Detection and Isolation of DNA from Metabolically Active Bacteria|journal=Applied and Environmental Microbiology|volume=65|issue=3|pages=1207–1213|doi= 10.1128/AEM.65.3.1207-1213.1999|date=March 1999|pmid=10049885|pmc=91166|bibcode=1999ApEnM..65.1207U |url=}}</ref> Subsequent sequencing of the labeled DNA can then be used to identify the microbial taxa that participated in the degradation of the added carbon source.

However, it is not certain whether all microbes present in an environmental sample can incorporate BrdU into their biomass during ''de novo'' DNA synthesis. Therefore, a group of microorganisms may respond to a carbon source but go undetected using this technique. Additionally, this technique is biased towards identifying microorganisms with A- and T-rich genomes.

DNA with BrdU transcribes as usual DNA, with [[guanine]] included in RNA as a complement to BrdU.<ref>{{cite journal |vauthors=Hill BT, Baserga R |title=Effect of 5-bromodeoxyuridine on the transcriptional properties of the genome in WI-38 human diploid fibroblasts |journal=Chem. Biol. Interact. |volume=10 |issue=5 |pages=363–375 |year=1975 |pmid=1095238 |doi= 10.1016/0009-2797(75)90058-7|bibcode=1975CBI....10..363H }}</ref>

* [[5-Bromouridine]]

* [[5-Ethynyl-2'-deoxyuridine]]

* [[Trypan blue]]

==References==

{{Reflist|2}}

* [http://openwetware.org/wiki/Bromodeoxyuridine_%28BrdU%29 BrdU at OpenWetWare]

* [http://www.idtdna.com/catalog/Modifications/Modifications.aspx?ProductID=1390 BrdU Modifications at IDT DNA]

[[Category:Genetics techniques]]

[[Category:Hydroxymethyl compounds]]