Genomic integrity of the Y chromosome sequence-tagged-sites in infertile and Down syndrome Jordanian males

Flow cytometric separation of X- and Y-chromosome bearing bovine sperm is an accepted technology for use at the commercial level. Nevertheless it is important to continue researching the area of gender-preselected sperm for improved efficiencies. We used a synthetic DNA mimic conjugated to a fluorescent dye for in situ detection of Y chromosomes in metaphase preparations of bovine somatic cells and spermatozoa. Peptide nucleic acids (PNA) are a type of DNA mimic having a higher affinity and stability than conventional DNA probes and are used as hybridization probes to complementary DNA. Using male bovine somatic cells and the Y-chromosome as a template, we arranged for the synthesis of a CY3-conjugated PNA to bind 13 to 15 base pairs of unique, Y-chromosome sequence. By testing different labeling conditions, we found that brief incubation (~1 h) of metaphase chromosomes with the PNA produced a localized signal on the Y-chromosome. No signals were observed when chromosomes of female bovine somatic cells were incubated with the same PNA probe. Because chromosomes occupy non-random territories in all cell nuclei, including sperm, we proposed to find centrally-located signals in 50% of fixed bovine sperm when treated with the same PNA as used for the somatic cells. As expected, we found the PNA signals present in 50% sperm (23/43) existing as a single, centrally-located, round fluorescent dot in the sperm head. Validation studies were also conducted using bovine sperm previously flow sorted into X or Y populations, and we found the signals in accordance to an expected signal present using the PNA (146/165 or 88.5% with PNA signal in presorted Y sperm heads and 13/174 or 7.5% with PNA signal in presorted X sperm heads).

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Gene discovery and comparative analysis of X-degenerate genes from the domestic cat Y chromosome☆☆Sequence data from this article have been deposited with the EMBL/GenBank Data Libraries under Accession No. EU879967-EU879988.

Genomics ◽

10.1016/j.ygeno.2008.06.012 ◽

2008 ◽

Vol 92 (5) ◽

pp. 329-338 ◽

Cited By ~ 36

Author(s):

Alison J. Pearks Wilkerson ◽

Terje Raudsepp ◽

Tina Graves ◽

Derek Albracht ◽

Wesley Warren ◽

...

Keyword(s):

Comparative Analysis ◽

Y Chromosome ◽

Sequence Data ◽

Gene Discovery ◽

Domestic Cat ◽

Chromosome Sequence ◽

Data Libraries

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Chromosomal abnormalities and Y chromosome microdeletions in infertile men with azoospermia and oligozoospermia in Eastern China

Journal of International Medical Research ◽

10.1177/0300060519896712 ◽

2019 ◽

Vol 48 (4) ◽

pp. 030006051989671

Author(s):

Jing Sha ◽

Guiping Huang ◽

Bei Zhang ◽

Xia Wang ◽

Zaochun Xu ◽

...

Keyword(s):

Y Chromosome ◽

Assisted Reproduction ◽

Sex Chromosome ◽

Chromosomal Abnormalities ◽

Eastern China ◽

Peripheral Blood Lymphocytes ◽

Specific Sequence ◽

Sequence Tagged Sites ◽

Infertile Men ◽

Y Chromosome Microdeletions

Objective The objective was to investigate the frequency and type of chromosomal abnormalities and Y chromosome microdeletions in infertile men with azoospermia and oligozoospermia to ensure appropriate genetic counseling before assisted reproduction in Eastern China. Methods A total of 201 infertile men (148 with azoospermia and 53 with oligozoospermia) were enrolled. Real-time PCR using six Y-specific sequence-tagged sites of the azoospermia factor (AZF) region was performed to screen for microdeletions. Karyotype analyses were performed on peripheral blood lymphocytes with standard G-banding. Results Out of 201 infertile patients, 22 (10.95%) had Y microdeletions [17/148 (11.49%) men with azoospermia and 5/53 (9.43%) men with oligozoospermia]. The most frequent microdeletions were in the AZFc region, followed by the AZFa+b + c, AZFb+c, AZFa, and AZFb regions. Chromosomal abnormalities were detected in 18.91% (38/201) of patients, 34 of which were sex chromosome abnormalities (16.92%) and 4 of which were autosomal abnormalities (1.99%). Chromosomal abnormalities were more prevalent in men with azoospermia (22.97%) than in those with oligozoospermia (7.55%). Conclusions We detected a high incidence of chromosomal abnormalities and Y chromosomal microdeletions in infertile Chinese men with azoospermia and oligozoospermia. These findings suggest the need for genetic testing before the use of assisted reproduction techniques.

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The Red Fox Y-Chromosome in Comparative Context

Genes ◽

10.3390/genes10060409 ◽

2019 ◽

Vol 10 (6) ◽

pp. 409 ◽

Cited By ~ 1

Author(s):

Halie M. Rando ◽

William H. Wadlington ◽

Jennifer L. Johnson ◽

Jeremy T. Stutchman ◽

Lyudmila N. Trut ◽

...

Keyword(s):

Y Chromosome ◽

Canis Lupus ◽

Related Species ◽

Red Fox ◽

Draft Genome ◽

Canis Lupus Familiaris ◽

Short Read ◽

Repeat Content ◽

Y Chromosomes ◽

Chromosome Sequence

While the number of mammalian genome assemblies has proliferated, Y-chromosome assemblies have lagged behind. This discrepancy is caused by biological features of the Y-chromosome, such as its high repeat content, that present challenges to assembly with short-read, next-generation sequencing technologies. Partial Y-chromosome assemblies have been developed for the cat (Felis catus), dog (Canis lupus familiaris), and grey wolf (Canis lupus lupus), providing the opportunity to examine the red fox (Vulpes vulpes) Y-chromosome in the context of closely related species. Here we present a data-driven approach to identifying Y-chromosome sequence among the scaffolds that comprise the short-read assembled red fox genome. First, scaffolds containing genes found on the Y-chromosomes of cats, dogs, and wolves were identified. Next, analysis of the resequenced genomes of 15 male and 15 female foxes revealed scaffolds containing male-specific k-mers and patterns of inter-sex copy number variation consistent with the heterogametic chromosome. Analyzing variation across these two metrics revealed 171 scaffolds containing 3.37 Mbp of putative Y-chromosome sequence. The gene content of these scaffolds is consistent overall with that of the Y-chromosome in other carnivore species, though the red fox Y-chromosome carries more copies of BCORY2 and UBE1Y than has been reported in related species and fewer copies of SRY than in other canids. The assignment of these scaffolds to the Y-chromosome serves to further characterize the content of the red fox draft genome while providing resources for future analyses of canid Y-chromosome evolution.

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