Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH Approach

Chromosomal instability in Alzheimer’s disease (AD) neurons has been previously reported. This pilot study aimed to establish a quantitative technique for assessing X chromosome centromere signals using fluorescence in situ hybridization (FISH). Hippocampal brain tissue was collected at autopsy from...

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Main Authors: Biljana Spremo-Potparević, Petar Popović, Dijana Topalović, Andrea Pirković, George Perry, Lada Živković
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Current Issues in Molecular Biology
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Online Access:https://www.mdpi.com/1467-3045/47/6/420
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author Biljana Spremo-Potparević
Petar Popović
Dijana Topalović
Andrea Pirković
George Perry
Lada Živković
author_facet Biljana Spremo-Potparević
Petar Popović
Dijana Topalović
Andrea Pirković
George Perry
Lada Živković
author_sort Biljana Spremo-Potparević
collection DOAJ
description Chromosomal instability in Alzheimer’s disease (AD) neurons has been previously reported. This pilot study aimed to establish a quantitative technique for assessing X chromosome centromere signals using fluorescence in situ hybridization (FISH). Hippocampal brain tissue was collected at autopsy from sporadic AD patients and age- and gender-matched controls. FISH was utilized to detect and measure the intensity of hybridization signals for X chromosome centromeres in the interphase nuclei of hippocampal brain cells. The premature centromere division (PCD) phenomenon, marked by a close bipartite signal appearing as two separated FISH spots, was examined to see if the hybridized DNA amount in each spot matched the expected centromere DNA amount. The technique effectively distinguished between PCD+ and PCD− signals. The average PCD frequency of the X chromosome in the AD group was 7 ± 1%, compared with 3.2 ± 0.84% in the controls. This quantitative approach supports qualitative analyses of FISH centromere spots, reinforcing findings of chromosomal instability in AD. The presence of a double signal at the centromere of a single X chromosome indicates re-entered cell cycles, DNA replication, and PCD in hippocampal neurons. This technique provides a reliable method for identifying PCD + signals and contributes to understanding chromosomal instability in AD.
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spelling doaj-art-abb618ac0a2a42c081cbaeaf26e985f02025-08-20T03:27:18ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452025-06-0147642010.3390/cimb47060420Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH ApproachBiljana Spremo-Potparević0Petar Popović1Dijana Topalović2Andrea Pirković3George Perry4Lada Živković5Department of Pathobiology, Faculty of Pharmacy, University of Belgrade, 11000 Belgrade, SerbiaDepartment of Pathobiology, Faculty of Pharmacy, University of Belgrade, 11000 Belgrade, SerbiaDepartment of Pathobiology, Faculty of Pharmacy, University of Belgrade, 11000 Belgrade, SerbiaDepartment of Biology of Reproduction, Institute for the Application of Nuclear Energy, 11000 Belgrade, SerbiaCollege of Sciences, The University of Texas, UTSA, San Antonio, TX 78249, USADepartment of Pathobiology, Faculty of Pharmacy, University of Belgrade, 11000 Belgrade, SerbiaChromosomal instability in Alzheimer’s disease (AD) neurons has been previously reported. This pilot study aimed to establish a quantitative technique for assessing X chromosome centromere signals using fluorescence in situ hybridization (FISH). Hippocampal brain tissue was collected at autopsy from sporadic AD patients and age- and gender-matched controls. FISH was utilized to detect and measure the intensity of hybridization signals for X chromosome centromeres in the interphase nuclei of hippocampal brain cells. The premature centromere division (PCD) phenomenon, marked by a close bipartite signal appearing as two separated FISH spots, was examined to see if the hybridized DNA amount in each spot matched the expected centromere DNA amount. The technique effectively distinguished between PCD+ and PCD− signals. The average PCD frequency of the X chromosome in the AD group was 7 ± 1%, compared with 3.2 ± 0.84% in the controls. This quantitative approach supports qualitative analyses of FISH centromere spots, reinforcing findings of chromosomal instability in AD. The presence of a double signal at the centromere of a single X chromosome indicates re-entered cell cycles, DNA replication, and PCD in hippocampal neurons. This technique provides a reliable method for identifying PCD + signals and contributes to understanding chromosomal instability in AD.https://www.mdpi.com/1467-3045/47/6/420Alzheimer’s diseasepremature centromere divisionX chromosomechromosomal instabilityfluorescence in situ hybridization
spellingShingle Biljana Spremo-Potparević
Petar Popović
Dijana Topalović
Andrea Pirković
George Perry
Lada Živković
Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH Approach
Current Issues in Molecular Biology
Alzheimer’s disease
premature centromere division
X chromosome
chromosomal instability
fluorescence in situ hybridization
title Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH Approach
title_full Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH Approach
title_fullStr Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH Approach
title_full_unstemmed Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH Approach
title_short Assessment of X Chromosome Centromere Instability in Alzheimer’s Disease: A Quantitative FISH Approach
title_sort assessment of x chromosome centromere instability in alzheimer s disease a quantitative fish approach
topic Alzheimer’s disease
premature centromere division
X chromosome
chromosomal instability
fluorescence in situ hybridization
url https://www.mdpi.com/1467-3045/47/6/420
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