High resolution R-bands produced in equine chromosomes after incorporation of bromodeoxyuridine.
Abstract: Cell synchronization was used to obtain an adequate percentage of very long chromosomes in equine mitotic spreads. Reported here is our variation, adapted to horse chromosomes, of a method using excess thymidine followed by bromodeoxyuridine incorporation. This technique routinely yields excellent quality cells, predominantly in prometaphase and prophase. Among other differences with the standard technique, this method does not use Colcemid, which, in addition to inhibiting spindle fiber formation, also increases chromosome contraction resulting in thicker and thus fewer bands. Consequently, horse prometaphase chromosomes, which have incorporated BrdU in the late-S-phase, are very long and display a large number of R-bands after the fluorescence-photolysis-Giemsa method. This technique should definitely be useful for the analysis of structural anomalies and the standardization of equine R-bands.
Publication Date: 1985-09-01 PubMed ID: 4056371
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research presents a modified method of producing high-resolution R-bands in horse chromosomes. The technique involves cell synchronization to procure long chromosomes and employing an adapted process that integrates bromodeoxyuridine after excess thymidine. The technique could serve as a useful tool in understanding structural anomalies and standardizing equine R-bands.
Cell Synchronization and Chromosome Elongation
- The research started with cell synchronization, a method that allows the simultaneous progression of cells through the cell cycle. This allowed the researchers to obtain a significant proportion of very long chromosomes in equine mitotic spreads.
Thymidine and Bromodeoxyuridine Incorporation Approach
- The researchers developed a new version tailoring a technique that employs excess thymidine followed by bromodeoxyuridine (bromo-deoxy-uridine or BrdU) incorporation to horse chromosomes. BrdU is an analog of thymidine and is often used to track DNA replication.
- The modified method resulted in yielding cells of excellent quality, mainly in the prometaphase and prophase, which are the last stages of prophase in eukaryotic cell division.
Comparative Differences from the Standard Technique
- The new technique doesn’t use Colcemid, a drug generally used to arrest dividing cells in metaphase by inhibiting spindle fiber formation. The use of Colcemid often causes the chromosomes to contract, leading to thicker and fewer R-bands, unfavorable for the research’s aim.
- Due to the absence of Colcemid, horse chromosomes that incorporated BrdU in the late-S-phase were visibly longer and exhibited a higher count of R-bands upon utilizing the fluorescence photolysis Giemsa method. Fluorescence photolysis Giemsa is a technique commonly used to create a specific banding pattern (R-banding) on chromosomes.
Potential Application of the Modified Technique
- The new technique standardized for equine chromosomes will be beneficial in analyzing structural anomalies in these chromosomes by visualizing more bands.
- The enhanced band count could aid in detailed genetic analysis and further diagnostic applications.
Cite This Article
APA
Romagnano A, Richer CL.
(1985).
High resolution R-bands produced in equine chromosomes after incorporation of bromodeoxyuridine.
J Hered, 76(5), 377-378.
Publication
Researcher Affiliations
MeSH Terms
- Animals
- Bromodeoxyuridine / pharmacology
- Cell Cycle / drug effects
- Cells, Cultured
- Chromosome Banding
- Chromosomes / drug effects
- Chromosomes / ultrastructure
- Horses / genetics
- Karyotyping
- Metaphase / drug effects
- Mitosis / drug effects
Citations
This article has been cited 2 times.- Iannuzzi L, Di Berardino D. Tools of the trade: diagnostics and research in domestic animal cytogenetics. J Appl Genet 2008;49(4):357-66.
- Richard F, Lombard M, Dutrillaux B. Reconstruction of the ancestral karyotype of eutherian mammals. Chromosome Res 2003;11(6):605-18.
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