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Analysis of sperm chromosomes in six carriers of rare and common Robertsonian translocations*

Postępy Higieny i Medycyny Doświadczalnej
Volume 75 (2021): Issue 1 (January 2021)
By:
Open Access
|Mar 2021

Figures & Tables

Fig. 1

Characteristics of Robertsonian translocations. A. Three types of rearrangement according to the breakpoints’ localization, leading to dicentric (left panel) or monocentric (middle and right panels) or derivative chromosome. B. Frequencies of rare and common rearrangements (marked with green or blue colour), and involvement of homo- or heterologous chromosomes (orange or yellow). C. Characteristics of chromosomes involved in rare Robertsonian translocations analyzed in this study: 45,XY,der(13;15)(q10;q10) mat (FISH with probes: centromere specific 13/21 (red) and 15 (green)); 45,XY,der(13;22)(q10;q10)mat (FISH with probes: centromere specific 13/21 (red) and 14/22 (green)). Microscope used: Olympus BX41, oil-immersed objective 100×, fluorescent filter-set: FITC/TexasRed/Triple/DAPI; software: ISIS (FISH) (MetaSystems, Germany)
Characteristics of Robertsonian translocations. A. Three types of rearrangement according to the breakpoints’ localization, leading to dicentric (left panel) or monocentric (middle and right panels) or derivative chromosome. B. Frequencies of rare and common rearrangements (marked with green or blue colour), and involvement of homo- or heterologous chromosomes (orange or yellow). C. Characteristics of chromosomes involved in rare Robertsonian translocations analyzed in this study: 45,XY,der(13;15)(q10;q10) mat (FISH with probes: centromere specific 13/21 (red) and 15 (green)); 45,XY,der(13;22)(q10;q10)mat (FISH with probes: centromere specific 13/21 (red) and 14/22 (green)). Microscope used: Olympus BX41, oil-immersed objective 100×, fluorescent filter-set: FITC/TexasRed/Triple/DAPI; software: ISIS (FISH) (MetaSystems, Germany)

Fig. 2

Schematic representation of meiotic segregation pattern of chromosomes observed in spermatozoa of Robertsonian translocation carriers, including trivalent configuration of chromosomes involved in rearrangements. Two different suggestions of FISH staining were presented: A. Whole chromosome painting (wcp) probes, that allow to differentiate normal and balanced gametes separately (different FISH phenotype; alternate segregants). B. Combination of probes specific for centromeres, subtelomeres, and chromosomal band (mostly used), but resulting in identical FISH phenotype for normal and balanced spermatozoa (alternate segregants)
Schematic representation of meiotic segregation pattern of chromosomes observed in spermatozoa of Robertsonian translocation carriers, including trivalent configuration of chromosomes involved in rearrangements. Two different suggestions of FISH staining were presented: A. Whole chromosome painting (wcp) probes, that allow to differentiate normal and balanced gametes separately (different FISH phenotype; alternate segregants). B. Combination of probes specific for centromeres, subtelomeres, and chromosomal band (mostly used), but resulting in identical FISH phenotype for normal and balanced spermatozoa (alternate segregants)

Fig. 3

GTG staining results for 45,XY,der(13;15)(q10;q10)mat carrier (R1). Microscope used: Olympus BX41, oil-immersed objective 100×, software: Ikaros (GTG) (Meta Systems, Germany). A. Whole metaphase plate. B. Chromosomes involved in the rearrangement
GTG staining results for 45,XY,der(13;15)(q10;q10)mat carrier (R1). Microscope used: Olympus BX41, oil-immersed objective 100×, software: Ikaros (GTG) (Meta Systems, Germany). A. Whole metaphase plate. B. Chromosomes involved in the rearrangement

Fig. 4

An example of FISH staining with whole chromosome painting (wcp) probes for 45,XY,der(13;15)(q10;q10) (R4) carrier. Probes used: 13 – red, 14 – green (Cytocell, UK). Microscope used: Olympus BX41, oil-immersed objective 100×, fluorescent filter-set: FITC/Texas Red/Triple/DAPI; software: ISIS (FISH) (Meta Systems, Germany)
An example of FISH staining with whole chromosome painting (wcp) probes for 45,XY,der(13;15)(q10;q10) (R4) carrier. Probes used: 13 – red, 14 – green (Cytocell, UK). Microscope used: Olympus BX41, oil-immersed objective 100×, fluorescent filter-set: FITC/Texas Red/Triple/DAPI; software: ISIS (FISH) (Meta Systems, Germany)

The result of meiotic segregation pattern analysis from the spermatozoa of rob(13;15) carrier (R1)_ Whole chromosome painting (wcp) probes of 2D-FISH were used, that allow to differentiate normal and balanced gametes separately (different FISH phenotype)

Segregation typeSchemesFISH patternprobes: 13 wcp red; 15 wcp greenSperm genotype%Total %
2:1Alternatenormal/balancedBALANCED2340.2675.96
22,–13,–15, +der(13;1535.70
Adjacent-1UNBALANCED23,–15, +der(13;15)8.0822.84
22, –133.76
Adjacent-223,-13, +der(13;15)6.37
22, –154.63
3:0 and/or 2n24, +der(13,15)1.091.09
Sum of unbalanced23.93
Unexplained signals0.11

Individual aneuploidy results from the spermatozoa of six Robertsonian translocation carriers (R1–R6) analyzed in the study, and our laboratory mean control value_ The results concern only the hyperhaploidy of chromosomes that are not involved in a particular translocation_ Grey color indicates results statistically higher than mean control value

No.KaryotypeFrequency of spermatozoa with disomy of chromosomes (n = 24) [%]:Frequency of diploid spermatozoa (2n) [%]
+7+9+18+21+22XXYYXY
Rare:
  R1rob(13;15)0.54*0.43*0.130.44*0.24*0.130.100.100.44*
  R2rob(13;15)0.34*0.35*0.120.29*0.29*0.120.090.121.06*
  R3rob(13;22)0.170.170.73*0.31*–0.160.22*0.33*0.84*
Common:
  R4rob(13;14)0.130.130.070.080.16*0.30*0.03*0.030.24*
  R5rob(13;14)ndnd0000.32*0.110.110
  R6rob(13;14)0.32*0.26*0.12000.100.100.110.06
Mean control value [n=7] ± SD#0.13±0.070.12±0.080.09±0.050.11±0.070.08±0.060.11±0.090.10±0.050.08±0.020.07±0.02

Semen assessment of the six Robertsonian translocation carriers (R1-R6)

No.KaryotypeSpermiogram (according to WHO, 2010 [88])Reproductive history
R145,XY,rob(13;15)(q10;q10)matOAT** data unavailable2 early miscarriages
R2*45,XY,rob(13;15)(q10;q10)matOA***: concentration: 3.2; motility: <30; normal forms: 9lack of conception
R3*45,XY,rob(13;22)(q10;q10)matOA: concentration 2.0; motility : 26; normal forms >14lack of conception
R4*45,XY,rob(13;14)(q10;q10)OA: concentration: 8.3; motility: 24; normal forms: 5.5lack of conception
R545,XY,rob(13;14)(q10;q10)OA: concentration: 0.7; motility: 24; normal forms: 0lack of conception
R645,XY,rob(13;14)(q10;q10)OA: concentration: 0.5; motility: 17; normal forms: 0twins after ICSI
WHO, 2010 [88]46,XYConcentration (106/ml): ≥15; Progressive motility (%): ≥32: Normal forms (%): ≥ 4
DOI: https://doi.org/10.5604/01.3001.0014.8122 | Journal eISSN: 1732-2693
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Language: English
Page range: 199 - 210
Submitted on: Jul 1, 2020
Accepted on: Dec 11, 2020
Published on: Mar 18, 2021
Published by: Hirszfeld Institute of Immunology and Experimental Therapy
In partnership with: Paradigm Publishing Services
Publication frequency: 1 issue per year
Keywords:
rob(13;15),
rob(13;22),
rob(13;14),
sperm aneuploidy,
meiotic segregation pattern,
rare Robertsonian translocation,
RobT
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Medicine,
Basic medical science,
Immunology

© 2021 Marta Olszewska, Ewa Wiland, Elzbieta Wanowska, Nataliya Huleyuk, Vyacheslav B. Chernykh, Danuta Zastavna, Maciej Kurpisz, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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