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A functional eukaryotic chromosome must contain three elements: a centromere, telomeres, and numerous origins of replication.
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Polytene chromosomes are giant interphase chromosomes with several DNA strands placed side by side. They were discovered in the year 1881 by Balbiani in salivary glands, intestine, muscles, malpighian tubules, and hypoderm of larvae Chironomus plumosus. Hence, these are also called "Salivary gland chromosomes." These are found in insects of the order Diptera and Collembola; in certain organs of mammals; and synergids, antipodes of flowering plants. Polytene chromosomes are also...
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Live Imaging Characterization of Centromere Movements During Male Meiotic Prophase in Arabidopsis thaliana
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Active centromere and chromosome identification in fixed cell lines.

Thian T Beh1, Ruth N MacKinnon2, Paul Kalitsis1

  • 1Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, Melbourne, VIC 3052 Australia ; Department of Paediatrics, University of Melbourne, Royal Children's Hospital, Parkville, Melbourne, VIC 3052 Australia.

Molecular Cytogenetics
|March 25, 2016
PubMed
Summary

A new method using an anti-CENP-C antibody effectively detects active centromeres in fixed cells, crucial for diagnosing chromosome instability disorders. This advance improves the accuracy of centromere abnormality assessment in various genetic conditions.

Keywords:
CENP-ACENP-CCentromereDicentricDicentric chromosome assay (DCA)Fluorescence in situ hybridisation (FISH)Human erythroleukaemia (HEL) cell lineImmunofluorescenceMulticolour FISH (mFISH)Neocentromere

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Area of Science:

  • Cytogenetics
  • Molecular Biology
  • Cell Biology

Background:

  • The centromere is vital for accurate chromosome segregation during cell division.
  • Centromere abnormalities are linked to cancer and genetic disorders, necessitating reliable detection methods.
  • Existing methods for functional centromere detection are limited to freshly harvested cells, not standard fixed cytogenetic samples.

Purpose of the Study:

  • To identify antibodies capable of recognizing active centromere antigens in methanol-acetic acid fixed cells.
  • To develop a robust method for assessing centromere function in stored cytogenetic samples.

Main Methods:

  • Screening of active centromere protein antibodies against methanol-acetic acid fixed cells.
  • Evaluation of a rabbit monoclonal antibody against human CENP-C.
  • Comparison and combination of centromere protein immunofluorescence (CENP-IF), centromere fluorescence in situ hybridisation (cenFISH), and multicolour FISH (mFISH).

Main Results:

  • A rabbit monoclonal anti-CENP-C antibody successfully recognized active centromeres in methanol-acetic acid fixed cells.
  • The combined CENP-IF-cenFISH-mFISH method, utilizing the anti-CENP-C antibody, proved effective.
  • The method demonstrated utility in cancer cell lines with known centromere defects, such as neocentromeres and functional dicentrics.

Conclusions:

  • The CENP-IF-cenFISH-mFISH method with anti-CENP-C is proposed for integration into standard cytogenetic analyses.
  • This includes applications in dicentric chromosome assay (DCA), prenatal screening, and constitutional/cancer karyotyping.
  • The method offers a more accurate assessment of centromere abnormality status in chromosome instability disorders.