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

Marta Olszewska
Marta Olszewska
, 
Ewa Wiland
Ewa Wiland
, 
Elzbieta Wanowska
Elzbieta Wanowska
, 
Nataliya Huleyuk
Nataliya Huleyuk
, 
Vyacheslav B. Chernykh
Vyacheslav B. Chernykh
, 
Danuta Zastavna
Danuta Zastavna
 and 
Maciej Kurpisz
Maciej Kurpisz
   | Mar 18, 2021
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Article Category: Original Article
Published Online: Mar 18, 2021
Page range: 199 - 210
Received: Jul 01, 2020
Accepted: Dec 11, 2020
DOI: https://doi.org/10.5604/01.3001.0014.8122
Keywords
© 2021 Marta Olszewska et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

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 type Schemes FISH patternprobes: 13 wcp red; 15 wcp green Sperm genotype % Total %
2:1 Alternatenormal/balanced BALANCED 23 40.26 75.96
22,–13,–15, +der(13;15 35.70
Adjacent-1 UNBALANCED 23,–15, +der(13;15) 8.08 22.84
22, –13 3.76
Adjacent-2 23,-13, +der(13;15) 6.37
22, –15 4.63
3:0 and/or 2n 24, +der(13,15) 1.09 1.09
Sum of unbalanced 23.93
Unexplained signals 0.11

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. Karyotype Frequency of spermatozoa with disomy of chromosomes (n = 24) [%]: Frequency of diploid spermatozoa (2n) [%]
+7 +9 +18 +21 +22 XX YY XY
Rare:
  R1 rob(13;15) 0.54* 0.43* 0.13 0.44* 0.24* 0.13 0.10 0.10 0.44*
  R2 rob(13;15) 0.34* 0.35* 0.12 0.29* 0.29* 0.12 0.09 0.12 1.06*
  R3 rob(13;22) 0.17 0.17 0.73* 0.31* – 0.16 0.22* 0.33* 0.84*
Common:
  R4 rob(13;14) 0.13 0.13 0.07 0.08 0.16* 0.30* 0.03* 0.03 0.24*
  R5 rob(13;14) nd nd 0 0 0 0.32* 0.11 0.11 0
  R6 rob(13;14) 0.32* 0.26* 0.12 0 0 0.10 0.10 0.11 0.06
Mean control value [n=7] ± SD# 0.13±0.07 0.12±0.08 0.09±0.05 0.11±0.07 0.08±0.06 0.11±0.09 0.10±0.05 0.08±0.02 0.07±0.02

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

No. Karyotype Spermiogram (according to WHO, 2010 [88]) Reproductive history
R1 45,XY,rob(13;15)(q10;q10)mat OAT** data unavailable 2 early miscarriages
R2* 45,XY,rob(13;15)(q10;q10)mat OA***: concentration: 3.2; motility: <30; normal forms: 9 lack of conception
R3* 45,XY,rob(13;22)(q10;q10)mat OA: concentration 2.0; motility : 26; normal forms >14 lack of conception
R4* 45,XY,rob(13;14)(q10;q10) OA: concentration: 8.3; motility: 24; normal forms: 5.5 lack of conception
R5 45,XY,rob(13;14)(q10;q10) OA: concentration: 0.7; motility: 24; normal forms: 0 lack of conception
R6 45,XY,rob(13;14)(q10;q10) OA: concentration: 0.5; motility: 17; normal forms: 0 twins after ICSI
WHO, 2010 [88] 46,XY Concentration (106/ml): ≥15; Progressive motility (%): ≥32: Normal forms (%): ≥ 4
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