FISH and Chimps: Insights into Frequency and Distribution of Sperm Aneuploidy in Chimpanzees (Pan troglodytes)

Numerical chromosomal aberrations in sperm are considered to be a major factor in infertility, early pregnancy loss and syndromes with developmental and cognitive disabilities in mammals, including primates. Despite numerous studies in human and farm animals, the incidence and importance of sperm aneuploidies in non-human primate remains mostly undetermined. Here we investigated the incidence and distribution of sperm aneuploidy in chimpanzees (Pan troglodytes), the species closest to human. We identify evolutionary conserved DNA sequences in human and chimpanzee and selected homologous sub-telomeric regions for all chromosomes to build custom probes and perform sperm-FISH analysis on more than 10,000 sperm nuclei per chromosome. Chimpanzee mean autosomal disomy rate was 0.057 ± 0.02%, gonosomes disomy rate was 0.198% and the total disomy rate was 1.497%. The proportion of X or Y gametes was respectively 49.94% and 50.06% for a ratio of 1.002 and diploidy rate was 0.053%. Our data provide for the first time an overview of aneuploidy in non-human primate sperm and shed new insights into the issues of aneuploidy origins and mechanisms.

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Sperm chromosomal abnormalities and their contribution to human embryo aneuploidy

Biology of Reproduction ◽

10.1093/biolre/ioz125 ◽

2019 ◽

Vol 101 (6) ◽

pp. 1091-1101 ◽

Cited By ~ 6

Author(s):

Lorena Rodrigo ◽

Marcos Meseguer ◽

Emilia Mateu ◽

Amparo Mercader ◽

Vanessa Peinado ◽

...

Keyword(s):

Clinical Outcomes ◽

Recurrent Miscarriage ◽

Sperm Concentration ◽

Sperm Parameters ◽

Fish Analysis ◽

Potential Contribution ◽

Comparative Genome Hybridization ◽

Abnormal Sperm ◽

Sperm Aneuploidy ◽

Sperm Fish

Abstract In this work we reviewed 18 years of experience using fluorescence in situ hybridization (FISH) for sperm aneuploidy testing. We evaluated parameters associated with increased numerical sperm chromosome abnormalities and determined the male contribution to embryo aneploidies in terms of reproductive outcome by increased sperm aneuploidy. This retrospective study analyzed data from 2008 sperm samples of infertile males undergoing FISH analysis because of clinical history of repetitive implantation failure, recurrent miscarriage, impaired sperm parameters, or mixed causes. Sperm concentration was the only sperm parameter associated with FISH results—we observed a gradual increase of abnormal sperm FISH results in males with decreasing sperm concentration. However, a great proportion of normozoospermic males also showed increased sperm aneuploidies, suggesting that sperm parameters alone do not enable identification of a substantial proportion of infertile males at risk of sperm aneuploidies. Regarding reproductive outcomes, couples with normal sperm FISH results for the male had similar outcomes regardless of conventional in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), or preimplantation genetic testing for aneuploidies (PGT-A). However, couples with abnormal sperm FISH results for the male showed better clinical outcomes after PGT-A, suggesting a potential contribution of sperm to embryo aneuploidy. Moreover, PGT-A cycles showed better clinical outcomes when 24 chromosomes were analyzed by array comparative genome hybridization (aCGH) or next-generation sequencing (NGS) instead of only nine chromosomes analyzed by FISH. In conclusion, sperm FISH analysis offers clinical prognostic value to evaluate reproductive possibilities in infertile couples. Therefore, couples with abnormal sperm FISH results should be offered genetic counseling and presented with clinical options such as PGT-A.

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Isolation and physical localization of new chromosome-specific centromeric repeats in farm animals

Veterinární Medicína ◽

10.17221/5541-vetmed ◽

2012 ◽

Vol 51 (No. 5) ◽

pp. 224-231 ◽

Cited By ~ 10

Author(s):

A. Pauciullo ◽

S. Kubickova ◽

H. Cernohorska ◽

K. Petrova ◽

D. Di Berardino ◽

...

Keyword(s):

Dna Sequences ◽

Animal Species ◽

Farm Animal ◽

Farm Animals ◽

Chromosome 1 ◽

Fish Analysis ◽

Repeated Dna ◽

Porcine Chromosome ◽

Strong Hybridization ◽

Painting Probes

In this study, new classes of tandemly repeated DNA sequences in the centromeric regions of three farm animal species are reported. Pericentromeric regions of bovine chromosome X, porcine chromosome 14 and equine chromosome 1 were microdissected by laser and amplified by DOP-PCR. Painting probes showed strong hybridization signals on their corresponding centromeres. Specific DNA fragments were isolated by cloning and sequenced. For each species, high homology level was found among the sequences. The presence of specific repeats within each sequence allows us to consider them as monomeric units or parts of longer tandemly repeated monomeric units that generate a specific higher-order repeat. In this respect, the isolation of new centromeric sequences enriches the genetic map of these three species and represents a useful tool for FISH analysis, thus expanding our knowledge on centromeres of farm animal species.

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Analysis of Meiotic Segregation Pattern and Interchromosomal Effects in a Bull Heterozygous for a 3/16 Robertsonian Translocation

Cytogenetic and Genome Research ◽

10.1159/000494289 ◽

2018 ◽

Vol 156 (4) ◽

pp. 197-203 ◽

Cited By ~ 1

Author(s):

Harmonie Barasc ◽

Nathalie Mouney-Bonnet ◽

Clémence Peigney ◽

Anne Calgaro ◽

Clémence Revel ◽

...

Keyword(s):

Robertsonian Translocation ◽

Chromosomal Rearrangements ◽

Segregation Pattern ◽

Fish Analysis ◽

Meiotic Segregation ◽

Robertsonian Translocations ◽

Sperm Nuclei ◽

Meiotic Segregation Pattern ◽

First Time ◽

Low Rate

Robertsonian translocations are the most frequent chromosomal rearrangements detected in cattle. Here, we report on the detection of a new Robertsonian translocation between chromosomes BTA3 and BTA16. This rob(3;16) was dicentric, suggesting that its occurrence was recent. FISH analysis of decondensed sperm nuclei revealed a relatively low rate of unbalanced gametes produced by adjacent segregation (5.87%). In addition, and for the first time in bovines, a significant interchromosomal effect (ICE) was detected for 2 different autosomes: BTA17 (global disomy + nullisomy rate of 9%) and BTA20 (1.8%). These results suggest that ICE should be taken into consideration when assessing the putative effect of Robertsonian translocations on reproduction.

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TERRA: A Novel Biomarker of Embryo Quality and Art Outcome

Genes ◽

10.3390/genes12040475 ◽

2021 ◽

Vol 12 (4) ◽

pp. 475

Author(s):

Maria Santa Rocca ◽

Ludovica Dusi ◽

Andrea Di Nisio ◽

Erminia Alviggi ◽

Benedetta Iussig ◽

...

Keyword(s):

Telomere Length ◽

Dna Sequences ◽

Male Fertility ◽

Embryo Quality ◽

Assisted Reproductive Technologies ◽

Biological Clock ◽

Reproductive Technologies ◽

Age Related ◽

Non Coding Rna ◽

First Time

Telomeres are considered to be an internal biological clock, and their progressive shortening has been associated with the risk of age-related diseases and reproductive alterations. Over recent years, an increasing number of studies have focused on the association between telomere length and fertility, identifying sperm telomere length (STL) as a novel biomarker of male fertility. Although typically considered to be repeated DNA sequences, telomeres have recently been shown to also include a long non-coding RNA (lncRNA) known as TERRA (telomeric repeat-containing RNAs). Interestingly, males with idiopathic infertility show reduced testicular TERRA expression, suggesting a link between TERRA and male fertility. The aim of this study was to investigate the role of seminal TERRA expression in embryo quality. To this end, STL and TERRA expression were quantified by Real Time qPCR in the semen of 35 men who underwent assisted reproductive technologies (ART) and 30 fertile men. We found that TERRA expression in semen and STL was reduced in patients that underwent ART (both p < 0.001). Interestingly, TERRA and STL expressions were positively correlated (p = 0.010), and TERRA expression was positively associated with embryo quality (p < 0.001). These preliminary findings suggest a role for TERRA in the maintenance of sperm telomere integrity during gametogenesis, and for the first time, TERRA expression was found as a predictive factor for embryo quality in the setting of assisted reproduction.

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Correlation of methylation status in MTHFR promoter region with recurrent pregnancy loss

Journal of Genetic Engineering and Biotechnology ◽

10.1186/s43141-021-00147-w ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Mai Mahmoud Shaker ◽

Taghreed Abdelmoniem shalabi ◽

Khalda said Amr

Keyword(s):

Dna Methylation ◽

Dna Sequences ◽

Promoter Region ◽

Pregnancy Loss ◽

Recurrent Pregnancy Loss ◽

Methylation Status ◽

Control Group ◽

Case Group ◽

Methyl Radical ◽

Fertile Women

Abstract Background DNA methylation is an epigenetic process for modifying transcription factors in various genes. Methylenetetrahydrofolate reductase (MTHFR) stimulates synthesis of methyl radical in the homocysteine cycle and delivers methyl groups needed in DNA methylation. Furthermore, numerous studies have linked gene polymorphisms of this enzyme with a larger risk of recurrent pregnancy loss (RPL), yet scarce information is available concerning the association between epigenetic deviations in this gene and RPL. Hypermethylation at precise DNA sequences can function as biomarkers for a diversity of diseases. We aimed by this study to evaluate the methylation status of the promoter region of MTHFR gene in women with RPL compared to healthy fertile women. It is a case–control study. Hundred RPL patients and hundred healthy fertile women with no history of RPL as controls were recruited. MTHFR C677T was assessed by polymerase chain reaction-restriction fragment length polymorphism (RFLP). Quantitative evaluation of DNA methylation was performed by high-resolution melt analysis by real-time PCR. Results The median of percentage of MTHFR promoter methylation in RPL cases was 6.45 [0.74–100] vs. controls was 4.50 [0.60–91.7], P value < 0.001. In the case group, 57 hypermethylated and 43 normo-methylated among RPL patients vs. 40 hypermethylated and 60 normo-methylated among controls, P< 0.005. Frequency of T allele in C677T MTHFR gene among RPL patients was 29% vs. 23% among the control group; C allele vs. T allele: odds ratio (OR) = 1.367 (95% confidence interval (CI) 0.725–2.581). Conclusion Findings suggested a significant association between hypermethylation of the MTHFR promoter region in RPL patients compared to healthy fertile women.

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Enzootic Activity of Chlamydia in Farms Located in a Hotspot Area for Zoonosis Emergence

10.21203/rs.3.rs-1014196/v1 ◽

2021 ◽

Author(s):

Ezequiel Andres Vanderhoeven ◽

Jessica P. Mosmann ◽

Adrián Díaz ◽

Cecilia G. Cuffini

Keyword(s):

Bird Species ◽

Farm Animals ◽

Economic Losses ◽

Wild Mammals ◽

Specific Pcr ◽

Chlamydial Species ◽

Zoonotic Risk ◽

Two Samples ◽

In The Wild ◽

First Time

Abstract Chlamydias are obligated intracellular Gram-negative bacteria, considered important zoonotic pathogens, broadly present in several bird species and responsible for economic losses in animal production. We analyzed the presence of Chlamydial species with zoonotic risk in farm animals in a highly biodiverse area and with great human circulation, the Argentine, Brazil and Paraguay tri-border area. We surveyed nine farms in an area and nasally swabbed a total of 62 animals. DNA was extracted and specific PCR was performed to identify chlamydial species. We detected Chlamydia spp . in 6.5% (4/62) of the animals tested, positive samples belonged to cattle and none of them showed symptoms of respiratory disease nor had been diagnose with reproductive diseases. Specific nested PCR confirmed two samples belonged to C. pecorum and two to C. psittaci . We report for the first time Chlamydia circulation with zoonotic risk in the region. Surveys in birds and wild mammals could give a better understanding to know what Chlamydial species are circulating in the wild interface. The zoonotic potential should be taking into account as farm workers and the surrounding population could be silent carriers or have respiratory diseases being underdiagnosed, and therefore should be considered in the differential diagnoses.

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O-40. Predictive value of sperm FISH analysis on the outcome of PGD for translocations

Reproductive BioMedicine Online ◽

10.1016/s1472-6483(12)60059-5 ◽

2002 ◽

Vol 4 ◽

pp. 28 ◽

Cited By ~ 1

Author(s):

T Escudero ◽

I Abdelhadi ◽

M Sandalinas ◽

S Munné

Keyword(s):

Predictive Value ◽

Fish Analysis ◽

Sperm Fish

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Characterization of Salt Overly Sensitive 1 (SOS1) gene homoeologs in quinoa (Chenopodium quinoa Willd.)

Genome ◽

10.1139/g09-041 ◽

2009 ◽

Vol 52 (7) ◽

pp. 647-657 ◽

Cited By ~ 60

Author(s):

P. J. Maughan ◽

T. B. Turner ◽

C. E. Coleman ◽

D. B. Elzinga ◽

E. N. Jellen ◽

...

Keyword(s):

Salt Tolerance ◽

Dna Sequences ◽

Genomic Sequence ◽

Chenopodium Quinoa ◽

Fish Analysis ◽

Cdna Sequences ◽

Grain Crop ◽

High Level ◽

Salt Overly Sensitive

Salt tolerance is an agronomically important trait that affects plant species around the globe. The Salt Overly Sensitive 1 (SOS1) gene encodes a plasma membrane Na+/H+ antiporter that plays an important role in germination and growth of plants in saline environments. Quinoa (Chenopodium quinoa Willd.) is a halophytic, allotetraploid grain crop of the family Amaranthaceae with impressive nutritional content and an increasing worldwide market. Many quinoa varieties have considerable salt tolerance, and research suggests quinoa may utilize novel mechanisms to confer salt tolerance. Here we report the cloning and characterization of two homoeologous SOS1 loci (cqSOS1A and cqSOS1B) from C. quinoa, including full-length cDNA sequences, genomic sequences, relative expression levels, fluorescent in situ hybridization (FISH) analysis, and a phylogenetic analysis of SOS1 genes from 13 plant taxa. The cqSOS1A and cqSOS1B genes each span 23 exons spread over 3477 bp and 3486 bp of coding sequence, respectively. These sequences share a high level of similarity with SOS1 homologs of other species and contain two conserved domains, a Nhap cation-antiporter domain and a cyclic-nucleotide binding domain. Genomic sequence analysis of two BAC clones (98 357 bp and 132 770 bp) containing the homoeologous SOS1 genes suggests possible conservation of synteny across the C. quinoa sub-genomes. This report represents the first molecular characterization of salt-tolerance genes in a halophytic species in the Amaranthaceae as well as the first comparative analysis of coding and non-coding DNA sequences of the two homoeologous genomes of C. quinoa.

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Towards a monophyletic classification of Lejeuneaceae III: the systematic position of Leiolejeunea

Phytotaxa ◽

10.11646/phytotaxa.170.3.4 ◽

2014 ◽

Vol 170 (3) ◽

pp. 187 ◽

Cited By ~ 13

Author(s):

ALFONS SCHÄFER-VERWIMP ◽

KATHRIN FELDBERG ◽

SHANSHAN DONG ◽

HUUB VAN MELICK ◽

DENILSON F. PERALTA ◽

...

Keyword(s):

Bayesian Inference ◽

Maximum Likelihood ◽

Dna Sequences ◽

Its Region ◽

Systematic Position ◽

Leaf Cell ◽

Oil Bodies ◽

Sister Relationship ◽

First Time

The derived liverwort Leiolejeunea grandiflora was recollected at the type locality in Jamaica after more than 100 years. The characteristics of its oil bodies were described for the first time based on the new collections. Each leaf cell possesses 2-4(-6) rather small, subhomogeneous to very finely segmented, subglobose to ellipsoidal, colorless oil bodies. The plants were either dioicous or autoicous. DNA sequences of two chloroplast regions (trnL-trnF, rbcL) and the nuclear ribosomal ITS region were obtained for two accessions of Leiolejeunea to enable the inference of the phylogenetic relationships of these plants. Based on Bayesian inference of phylogeny as well as maximum parsimony and maximum likelihood analyses of a dataset including 87 representatives of Lejeuneaceae, Leiolejeunea was found as the putative sister to either Echinolejeuneinae or Cheilolejeuneinae. Thus, we propose the new monogeneric subtribe Leiolejeuneinae with relationships to Cheilolejeuneinae and Echinolejeuneinae. The analyses included also one accession of the generitype of Cheilolejeunea, C. decidua [= Cheilolejeunea adnata]. This species was found in a well supported sister relationship with Cystolejeunea. To avoid nomenclatural confusion, we propose a wide genus concept for Cheilolejeunea including Aureolejeunea, Cyrtolejeunea, Cystolejeunea, Evansiolejeunea, Leucolejeunea, and Omphalanthus.

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A Familial Small Supernumerary Marker Chromosome 15 Associated with Cryptic Mosaicism with Two Different Additional Marker Chromosomes Derived de novo from Chromosome 9: Detailed Case Study and Implications for Recurrent Pregnancy Loss

Cytogenetic and Genome Research ◽

10.1159/000494822 ◽

2018 ◽

Vol 156 (4) ◽

pp. 179-184

Author(s):

Vida Čulić ◽

Ruzica Lasan-Trcić ◽

Thomas Liehr ◽

Igor N. Lebedev ◽

Maja Pivić ◽

...

Keyword(s):

Pregnancy Loss ◽

De Novo ◽

Marker Chromosome ◽

Chromosome 9 ◽

Chromosome 15 ◽

Normal Female ◽

Fish Analysis ◽

Spontaneous Abortions ◽

Small Supernumerary Marker Chromosome ◽

Trisomy 9

We report a case of familial small supernumerary marker chromosome 15 in a phenotypically normal female with 4 recurrent spontaneous abortions and a healthy child. The initial karyotype showed a small, bisatellited, apparently metacentric marker chromosome, 47,XX,+idic(15)(q11.1), maternally inherited. The proband's mother was mosaic for the idic(15)(q11.1) without pregnancy loss. Reexamination of the proband's karyotype revealed cryptic mosaicism for 1 ring and 1 minute chromosome derived de novo from chromosome 9 in 2% of the metaphases. In FISH analysis, the patient's karyotype was mos 47,XX,+idic(15)(q11.1)mat[100]/49,XX,+idic(15)(q11.1)mat,+r(9;9;9;9),+der(9)dn[2]. The second spontaneous abortion had trisomy 9 (47,XX,+9); the third had mosaic trisomy 9 in 21% of the nuclei and isodicentric chromosome 15 in 36% of the nuclei (mos 48,XN,+9,+idic(15)(q11.1)/47,XN,+9/47,XN,+idic(15)(q11.1)/46,XN). The first and fourth abortions were not cytogenetically studied. The cause of the spontaneous abortions in this patient is likely the cryptic mosaicism for ring and minute chromosomes 9, and gonadal mosaicism is most probable, due to the 2 abortions.

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