Dataset Information

Sperm Global DNA Methylation (SGDM) in Semen of Healthy Dogs.

ABSTRACT: Male infertility is an emerging problem in both humans and animals, and the knowledge of its causes is the first step to identifying new diagnostic and therapeutic strategies. In humans, alteration of sperm DNA methylation have been related to poor quality semen, impaired seminal parameters, azoospermia and reduced fertility. Although semen analysis is routinely used to evaluate the male reproductive potential in the canine species, no authors have attempted to relate semen characteristics to the sperm global DNA methylation (SGDM). The aim of this study was to evaluate the SGDM level in healthy dogs and to correlate it with semen parameters that are currently used in dog semen analyses. Conventional and unconventional (sperm DNA fragmentation and SGDM) seminal parameters of thirty dogs from different breeds were evaluated. A positive correlation was found between SGDM and sperm concentration (r = 0.41; p < 0.05), and total sperm count (r = 0.61; p < 0.001); SGDM was significantly lower in oligozoospermic vs non-oligozoospermic dogs (4.3% vs. 8.7%; p < 0.005). Our findings suggest that SGDM levels are related to conventional seminal parameters, and could be used as a marker of testis function and spermatogenesis in dogs.

SUBMITTER: Galdiero G

PROVIDER: S-EPMC8002840 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Sperm Global DNA Methylation (SGDM) in Semen of Healthy Dogs.

Galdiero Giacomo G D'Anza Emanuele E de Angelis Cristina C Albarella Sara S Peretti Vincenzo V Pivonello Rosario R Ciotola Francesca F

Veterinary sciences 20210317 3

Male infertility is an emerging problem in both humans and animals, and the knowledge of its causes is the first step to identifying new diagnostic and therapeutic strategies. In humans, alteration of sperm DNA methylation have been related to poor quality semen, impaired seminal parameters, azoospermia and reduced fertility. Although semen analysis is routinely used to evaluate the male reproductive potential in the canine species, no authors have attempted to relate semen characteristics to th ...[more]

PMID: 33802963

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Project description:Non-Hodgkin lymphomas are among the most common types of tumors in dogs, and they are currently accepted as comparative models of the disease in humans. Aberrant patterns of DNA methylation seem to play a key role in the development of hematopoietic neoplasms in humans, constitute a special mechanism of transcriptional control, and may be influenced by genetic and environmental factors. Blood leukocyte DNA global methylation has been poorly investigated in dogs. The aim of this study is to examine whether peripheral blood global DNA methylation is associated with canine multicentric lymphomas. Peripheral venous blood samples from ten healthy dogs and nine dogs bearing multicentric lymphomas were collected, and the buffy coat was separated. Global DNA methylation was analyzed by High Performance Liquid Chromatography (HPLC) and immunocytochemistry (ICC). In both analyses, leukocytes from dogs with lymphoma presented lower global DNA methylation than in healthy dogs (HPLC: p = 0.027/ 5MeCyt immunoreactivity scores: p = 0.015). Moderate correlation was observed between the results obtained by HPLC and ICC (correlation coefficient = 0.50). For the identification of differently methylated genes between both groups, the Infinium Human Methylation (HM) EPIC BeadChip (850K) was used. Of the 853,307 CpGs investigated in the microarray, there were 34,574 probes hybridized in the canine samples. From this total, significant difference was observed in the methylation level of 8433 regions, and through the homologous and orthologous similarities 525 differently methylated genes were identified between the two groups. This study is pioneer in suggesting that dogs bearing non-Hodgkin lymphoma presented DNA global hypomethylation of circulating leukocytes compared with healthy dogs. Although canine samples were used in an assay developed specifically for human DNA, it was possible to identify differently methylated genes and our results reiterate the importance of the use of peripheral blood leukocytes in cancer research and possible new biomarkers targets.

Project description:BackgroundAltered sperm DNA methylation patterns of imprinted genes as well as certain spermatogenesis-related genes has been proposed as a possible mechanism of male subfertility. Some reports suggest that there is an elevated risk of congenital diseases, associated with imprinted genes, in children conceived via intra-cytoplasmic sperm injection, due to the involvement of spermatozoa with aberrant imprinted genes obtained from infertile men.MethodsIn this study, the DNA methylation status of the promoter regions of six imprinted genes, namely potassium voltage-gated channel subfamily Q member 1 (KCNQ1), maternally expressed gene 3 (MEG3), insulin-like growth factor 2 (IGF-2), KCNQ1 overlapping transcript 1 (KCNQ1OT1), mesoderm specific transcript (MEST), and paternally expressed gene 3 (PEG3), were detected by a next generation sequencing-based multiple methylation-specific polymerase chain reaction analysis of sperm samples obtained from 166 men who sought fertility evaluation in our Reproductive Medicine Center. Thereafter, the semen samples were classified into subgroups according to sperm motility and DNA integrity status.ResultsAs compared to the normozoospermic group, the samples of the asthenospermic group exhibited significant hypermethylation in two CpG sites of IGF-2 and significant hypomethylation in one CpG site of KCNQ1 as well as three CpG sites of MEST (P < 0.05). However, we did not observe any significant differences in the overall methylation levels of these six imprinted genes (P > 0.05). Additionally, we found that 111 of 323 CpG sites were hypomethylated in the group with DNA fragmentation index (DFI) ≥ 30% as compared to the group with DFI < 30% (P < 0.05). In this case, there were significant differences in the overall methylation levels of MEG3, IGF-2, MEST, and PEG3 (P < 0.05), but not in that of KCNQ1OT1 and KCNQ1 (P > 0.05). Hence, aberrant methylation patterns of imprinted genes were more prevalent in males with poor sperm quality, especially in those with severe sperm DNA damage.ConclusionIn conclusion, abnormal DNA methylation of some CpG sites of imprinted genes are associated with poor sperm quality, including asthenospermia and severe sperm DNA impairment.

Dataset Information

Sperm Global DNA Methylation (SGDM) in Semen of Healthy Dogs.

Publications

Sperm Global DNA Methylation (SGDM) in Semen of Healthy Dogs.

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