A Role for Synaptonemal Complex in Meiotic Mismatch Repair

A large subset of meiotic recombination intermediates form within the physical context of synaptonemal complex (SC), but the functional relationship between SC structure and homologous recombination remains obscure. Our prior analysis of strains deficient for SC central element proteins demonstrated that tripartite SC is dispensable for interhomolog recombination in S. cerevisiae. Here we report that while dispensable for recombination per se, SC proteins promote efficient mismatch repair at interhomolog recombination sites. Failure to repair mismatches within heteroduplex-containing meiotic recombination intermediates leads to genotypically sectored colonies (post meiotic segregation events). We discovered increased post-meiotic segregation at THR1 in cells lacking Ecm11 or Gmc2, or in the SC-deficient but recombination-proficient zip1[Δ21-163] mutant. High-throughput sequencing of octad meiotic products furthermore revealed a genome-wide increase in recombination events with unrepaired mismatches in ecm11 mutants relative to wild type. Meiotic cells missing Ecm11 display longer gene conversion tracts, but tract length alone does not account for the higher frequency of unrepaired mismatches. Interestingly, the per-nucleotide mismatch frequency is elevated in ecm11 when analyzing all gene conversion tracts, but is similar between wild type and ecm11 if considering only those events with unrepaired mismatches. Thus, in both wild type and ecm11 strains a subset of recombination events is susceptible to a similar degree of inefficient mismatch repair, but in ecm11 mutants a larger fraction of events fall into this inefficient repair category. Finally, we observe elevated post-meiotic segregation at THR1 in mutants with a dual deficiency in MutSγ crossover recombination and SC assembly, but not in the mlh3 mutant, which lacks MutSγ crossovers but has abundant SC. We propose that SC structure promotes efficient mismatch repair of joint molecule recombination intermediates, and that absence of SC is the molecular basis for elevated post-meiotic segregation in both MutSγ crossover-proficient (ecm11, gmc2) and MutSγ crossover-deficient (msh4, zip3) strains.

P-525 Analysis of segregation patterns of trivalent structure and the effect on genome stability in Robertsonian translocation carriers

Study question What are the factors that affect the separation pattern of Robertsonian translocation trivalent, and whether the structure of the trivalent affected the chromosome stability? Summary answer The meiotic segregation modes can be affected by the carrier’s sex and special chromosome, and a trivalent structure can affect the stability of the genome. What is known already Robertson translocation occurs when two proximal acrocentric chromosomes fuse at the centromere, and forms a trivalent structure during meiosis. This structure will affect the fertility of Robertsonian translocation carriers, and may destroy the stability of the genome by affecting the separation of other chromosomes, which is called Inter-Chromosomal Effect (ICE). Previous research have confirmed that the use of PGT in Robertsonian translocation carriers can effectively reduce abortion and increase live birth. But some studies dispute this conclusion and the existence of ICE. However, there is no large data study to verify these controversies. Study design, size, duration PGT results of 928 oocyte retrieval cycles in 763 couples(one of the couples is a Robertsonian translocation carrier) were analysied from December 2012 to June 2020. A total of 1492 couples who received PGT-A were collected as control group, and matched according to age and testing time stage. The study was approved by the ethics committee(LL-SC-SG-2006-008 and LL-SC-SG-2014-016). Participants/materials, setting, methods Cytogenetic analysis was performed using GTG standard method (trypsin and GiemsaG banding) to analyze the chromosomes of peripheral blood lymphocytes. Blastocysts obtained by standard IVF procedure were biopsied on the 5th or 6th morning after fertilization, and the trophoblast cells were amplified by PicoPLEX whole genome amplification kit (Rubicon Genology) or Repli-g Single Cell Kit(Qiagen). PGT-SR was performed using SNP array or NGS as previously described. Main results and the role of chance In this study, a total of 3423 blastocysts from 763 couples were analysed using SNP-array or NGS. Among them, the rate of alternate segregation of male Robertsonian translocation carriers was significantly higher than that in female carriers (82.26% vs 59.96%, P < 0.001), and meiotic segregation modes could be affected by the special chromosome such as 13 in female(P = 0.042) and 15 in male(P = 0.045) involved. A trivalent structure can affect the stability of the genome during mitosis, which is associated with an increase in the propotion of chromosome mosaic compared with the PGT-A control group(1.18% vs 0.53%, P < 0.01). In addition, we found an interesting phenomenon: in the meiotic segregation of female Robertsonian translocation carriers associated with chromosomes 21 and 22, the chromosome 21 or 22 of the two chromosomes involved in translocation are more likely to be abnormal, and according to our results, the effect of chromosome 21 seems to be greater. Limitations, reasons for caution (1) Limitations of retrospective analysis; (2) The results are not fully representative of the general population; (3) PGT-A patients always had repeated implantation failure or recurrent aboration, which may cause deviation to the results. Wider implications of the findings This study analyzed the influencing factors of the separation patterns of trivalent, and verified the existence of ICE. This suggest that PGT-SR can have a better outcome in patients with Robertsonian translocation, especially in male carriers. These results will provide carrier couple with more appropriate genetic counseling. Trial registration number no

P–525 Analysis of segregation patterns of trivalent structure and the effect on genome stability in Robertsonian translocation carriers

Study question What are the factors that affect the separation pattern of Robertsonian translocation trivalent, and whether the structure of the trivalent affected the chromosome stability? Summary answer The meiotic segregation modes can be affected by the carrier’s sex and special chromosome, and a trivalent structure can affect the stability of the genome. What is known already Robertson translocation occurs when two proximal acrocentric chromosomes fuse at the centromere, and forms a trivalent structure during meiosis. This structure will affect the fertility of Robertsonian translocation carriers, and may destroy the stability of the genome by affecting the separation of other chromosomes, which is called Inter-Chromosomal Effect (ICE). Previous research have confirmed that the use of PGT in Robertsonian translocation carriers can effectively reduce abortion and increase live birth. But some studies dispute this conclusion and the existence of ICE. However, there is no large data study to verify these controversies. Study design, size, duration PGT results of 928 oocyte retrieval cycles in 763 couples(one of the couples is a Robertsonian translocation carrier) were analysied from December 2012 to June 2020. A total of 1492 couples who received PGT-A were collected as control group, and matched according to age and testing time stage. The study was approved by the ethics committee(LL-SC-SG–2006–008 and LL-SC-SG–2014–016). Participants/materials, setting, methods Cytogenetic analysis was performed using GTG standard method (trypsin and GiemsaG banding) to analyze the chromosomes of peripheral blood lymphocytes. Blastocysts obtained by standard IVF procedure were biopsied on the 5th or 6th morning after fertilization, and the trophoblast cells were amplified by PicoPLEX whole genome amplification kit (Rubicon Genology) or Repli-g Single Cell Kit(Qiagen). PGT-SR was performed using SNP array or NGS as previously described. Main results and the role of chance In this study, a total of 3423 blastocysts from 763 couples were analysed using SNP-array or NGS. Among them, the rate of alternate segregation of male Robertsonian translocation carriers was significantly higher than that in female carriers (82.26% vs 59.96%, P < 0.001), and meiotic segregation modes could be affected by the special chromosome such as 13 in female(P = 0.042) and 15 in male(P = 0.045) involved. A trivalent structure can affect the stability of the genome during mitosis, which is associated with an increase in the propotion of chromosome mosaic compared with the PGT-A control group(1.18% vs 0.53%, P < 0.01). In addition, we found an interesting phenomenon: in the meiotic segregation of female Robertsonian translocation carriers associated with chromosomes 21 and 22, the chromosome 21 or 22 of the two chromosomes involved in translocation are more likely to be abnormal, and according to our results, the effect of chromosome 21 seems to be greater. Limitations, reasons for caution (1) Limitations of retrospective analysis; (2) The results are not fully representative of the general population; (3) PGT-A patients always had repeated implantation failure or recurrent aboration, which may cause deviation to the results. Wider implications of the findings: This study analyzed the influencing factors of the separation patterns of trivalent, and verified the existence of ICE. This suggest that PGT-SR can have a better outcome in patients with Robertsonian translocation, especially in male carriers. These results will provide carrier couple with more appropriate genetic counseling. Trial registration number no

O-206 Meiotic segregation analysis for reciprocal translocation carriers: Assessment of factors influencing meiotic segregation patterns

Study question To analyze factors that could influence meiotic segregation patterns for reciprocal translocation carriers. Summary answer Involvement of an Acr-ch, female gender, and lower TAR1 (ratio of translocated segment 1 over the chromosome arm) were independent risk factors for alternate segregation. What is known already Reciprocal translocation is one of the more common structural rearrangements of chromosomes, which is associated with reproductive risks, such as infertility, spontaneous abortion and the delivery of babies with mental retardation or developmental delay. Extensive studies on meiotic segregation patterns of sperm, blastomere, and blastocysts have identified several factors that may influence the generation of unbalanced rearrangement of reciprocal translocations, including carrier’s gender and age, location of breakpoints, chromosome type, and the quadrivalent structure. However, some results are controversial. Study design, size, duration A retrospective study from October 2013 to December 2019, a total of 10846 blastocysts originating from 2871 oocyte retrieval cycles from 2253 couples with one of the partners carrying reciprocal were investigated. The mean maternal age was 29.97±4 years (20 –47years). Participants/materials, setting, methods Trophectoderm biopsy of blastocysts was performed on the 5th or 6th day of development. Whole genome amplification (WGA) was performed on all samples, and the WGA was analyzed with SNP array or NGS. Segregation patterns of quadrivalent in 10846 blastocysts were analyzed. Risk factors for segregation patterns were explored through analyzing carriers’ demographic and cytogenetic characteristics using multivariate generalized linear mixed models (GLMMs). Main results and the role of chance The percentage of normal/balanced blastocysts was 34.3%, and 2:2 segregation was observed in 90.0% of blastocysts. Increased TAR1 (the ratio of translocated segment 1 over the chromosome arm) was noted as an independent protective factor for the proportion of alternate segregation (P = 0.004). The female gender and involvement of an Acr-ch were found independent risk factors for alternate segregation (P < 0.001). A higher TAR1 reduced the risk of adjacent-1 segregation; longer translocated segment and female gender increased the risk of adjacent-2 segregation (P = 0.009 and P < 0.001, respectively). Female gender and involvement of an Acr-ch enhanced the risk of 3:1 segregation (P < 0.001 and P = 0.012, respectively). Limitations, reasons for caution About 1400 blastocysts were not diagnosed in the 2871 cycles, which might cause bias in the results. Secondly, the interchromosomal effect of reciprocal translocations was not analyzed in this study. Wider implications of the findings In conclusion, a carrier’s gender, involvement of an Acr-ch, and location of breakpoints may influence the segregation patterns. Besides, involvement of an Acr-ch, female gender, and lower TAR1 are independent risk factors for alternate segregation. These results may provide more appropriate genetic counseling for couples with balanced translocation. Trial registration number no

A Role for Synaptonemal Complex in Meiotic Mismatch Repair

During meiosis a large subset of interhomolog recombination repair intermediates form within the physical context of the synaptonemal complex (SC), a protein-rich structure assembled at the interface of aligned homologous chromosomes. However, the functional relationship between SC structure and homologous recombination remains poorly defined. In prior work we determined that tripartite SC is dispensable for recombination in S. cerevisiae; SC central element proteins Ecm11 and Gmc2 instead limit the number of recombination events. Here we report that while dispensable for recombination per se, SC central element proteins influence the processing of interhomolog recombination intermediates in a manner that minimizes errors in mismatch correction. Failure to correct mis-paired bases within heteroduplex at meiotic recombination sites leads to genotypically sectored colonies (post meiotic segregation events) arising from mitotic proliferation of mismatch-containing spores. We discovered an increase in post-meiotic segregation at the THR1 locus in cells lacking Ecm11 or Gmc2, or in the SC-deficient but crossover recombination-proficient zip1[Δ21-163] mutant. High-throughput sequencing of octad meiotic products revealed a genome-wide increase in recombination events with uncorrected mismatches in ecm11 mutants relative to wild type. Meiotic cells missing Ecm11 also display longer gene conversion tracts, but tract length alone does not account for the higher frequency of uncorrected mismatches. Interestingly, the per-nucleotide mismatch frequency is elevated in ecm11 mutants when analyzing all gene conversion tracts, but is similar between wild type and ecm11 if one considers only those events with uncorrected mismatches. Our data suggest that a subset of recombination events is similarly susceptible to mismatch repair errors in both wild type and ecm11 strains, but in ecm11 mutants many more recombination events fall into this inefficient repair category. Finally, we observe elevated post-meiotic segregation at THR1 in mutants with a dual deficiency in MutSγ-mediated crossover recombination and SC assembly, but not in the mlh3 mutant, which lacks MutSγ crossovers but has abundant SC. We propose that SC structure promotes efficient mismatch repair of joint molecule recombination intermediates resolved via both MutSγ-associated and MutSγ-independent pathways, and is the molecular basis for elevated post-meiotic segregation in both MutSγ crossover-proficient (ecm11, gmc2) and MutSγ crossover-deficient (msh4, zip3) strains.

Analysis of chromosome segregation in reciprocal translocation carriers

Аутосомные реципрокные транслокации (АРТ) приводят к повышенному риску образования несбалансированных гамет вследствие патологической сегрегации хромосом в мейозе у носителей. В настоящей статье приведены результаты анализа типов сегрегации для 26 АРТ, а также определены теоретически возможные варианты сегрегации хромосом. В 73% случаев у носителей АРТ в более, чем 50% бластомеров наблюдалось совпадение теоретического и детектируемого типов сегрегации. Полученные данные можно использовать для оптимизации персонализированного медико-генетического консультирования семей, где один из супругов является носителем АРТ, и имеющих репродуктивные проблемы, высокий риск неразвивающейся беременности и/или рождения ребенка с хромосомной патологией. Autosomal reciprocal translocations (ART) lead to an increased risk of imbalanced gametes formation due to pathological meiotic segregation. Segregation type was analyzed and theoretical segregation pattern was determined in 26 cleavage stage embryos in this article. A coincidence of theoretical and detectable segregation types was observed in more than 50 % of blastomeres in 73 % of cases. The data obtained may be used for personalized genetic counseling in families with high risks of recurrent spontaneous abortions, infertility or children with birth defects due to ART.