A novel approach to sexing bovine blastocysts using male-specific gene expression

Hamilton, Christine K.; Combe, Audrey; Caudle, Jeffrey; Ashkar, F. A.; Macaulay, Angus D.; Blondin, Patrick; King, W. A.

doi:10.1016/j.theriogenology.2011.11.027

Cited by 25 publications

(21 citation statements)

References 54 publications

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“…Presumptive zygotes were stripped of their cumulus cells by vortex after 18–20 h post‐insemination and then cultured in groups of 30 in 30 μl drops of synthetic oviduct fluid medium for 48 h. The zona pellucida was removed using 0.2% pronase from the collected 2‐ to 8‐cell stage embryos ( n = 100), then individually lysed and sexed by PCR amplifying the Y‐chromosome‐specific TSPY and autosomal control GAPDH genes according to Hamilton et al. ().…”

Section: Methodsmentioning

confidence: 99%

Enrichment of Y‐Chromosome‐Bearing Bull Spermatozoa by Swim‐up Through a Column

Azizeddin

et al. 2013

Reprod Domestic Animals

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With the advancement of assisted reproductive biotechnologies, preselecting the sex of offspring has become an important goal for cattle and other livestock breeding as well as for research. The aim of this study was to investigate the feasibility of producing enriched pools of X- or Y-chromosome-bearing sperm by vertical swim-up through a long, narrow column. Sperm recovered from the top portion of the column was predominantly Y-bearing (60%, p < 0.05), which were capable of fertilizing matured oocytes and produce significantly more male embryos compared with standard swim-up protocol.

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Section: Methodsmentioning

confidence: 99%

Enrichment of Y‐Chromosome‐Bearing Bull Spermatozoa by Swim‐up Through a Column

Azizeddin

et al. 2013

Reprod Domestic Animals

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“…To check the accuracy of sperm selection, the sexes of single blastocysts made using X-sorted or Y-sorted sperm were determined by the PCR for DDX3Y, USP9Y, and ZRSR2Y on genomic DNA from blastocysts. Control experiments indicated that sorted sperm was indeed suitable for generation of sex-specific embryos (Table S3) [34]. Here, we focused on the period between the 8-cell stage and the blastocyst, since we found most changes in XCI related gene expression to arise after the 8-cell stage (Figure 1).…”

Section: Gene Expression Differences Between Male and Female Embryosmentioning

confidence: 99%

Initiation of X Chromosome Inactivation during Bovine Embryo Development

Tol

Stout

et al. 2020

Cells

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X-chromosome inactivation (XCI) is a developmental process that aims to equalize the dosage of X-linked gene products between XY males and XX females in eutherian mammals. In female mouse embryos, paternal XCI is initiated at the 4-cell stage; however, the X chromosome is reactivated in the inner cell mass cells of blastocysts, and random XCI is subsequently initiated in epiblast cells. However, recent findings show that the patterns of XCI are not conserved among mammals. In this study, we used quantitative RT-PCR and RNA in situ hybridization combined with immunofluorescence to investigate the pattern of XCI during bovine embryo development. Expression of XIST (X-inactive specific transcript) RNA was significantly upregulated at the morula stage. For the first time, we demonstrate that XIST accumulation in bovine embryos starts in nuclei of female morulae, but its colocalization with histone H3 lysine 27 trimethylation was first detected in day 7 blastocysts. Both in the inner cell mass and in putative epiblast precursors, we observed a proportion of cells with XIST RNA and H3K27me3 colocalization. Surprisingly, the onset of XCI did not lead to a global downregulation of X-linked genes, even in day 9 blastocysts. Together, our findings confirm that diverse patterns of XCI initiation exist among developing mammalian embryos.

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“…In relation to these findings, the most interesting question is the potential consequences of the detected extensive somatic variability of a gene whose expression is restricted to the adult testis. However, the recently found TSPY expression in early bovine embryos [Hamilton et al, 2012b] suggests a yet unknown function in males. TSPY copy number variability in early embryonic divisions not only leads to somatic mosaicism but could also contribute to variable success of embryo development.…”

Section: Methodsmentioning

confidence: 96%

Somatic Mosaicism in Bulls Estimated from Genome-Wide CNV Array and <b><i>TSPY</i></b> Gene Copy Numbers

Oluwole

Révay²,

Mahboubi³

et al. 2016

Cytogenet Genome Res

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Somatic mosaicism has become a focus in human research due to the implications of individual genetic variability in disease. Here, we assessed somatic copy number variations (CNVs) in Holstein bulls in 2 respects. We estimated genome-wide CNVs and assayed CNVs of the TSPY gene, the most variable bovine gene from the Y chromosome. Somatic tissues (blood, lung, heart, muscle, testis, and brain) of 4 bulls were arrayed on the Illumina Bovine SNP50k chip and qPCR tested for TSPY copy numbers. Our results showed extensive copy number divergence in tissues within the same animal as well as significant copy number alterations of TSPY. We detected a mean of 31 CNVs per animal among which 14 were of germline origin, as they were constantly present in all investigated tissues of the animal, while 18 were specific to 1 tissue. Thus, 57% of the total number of detected CNVs was the result of de novo somatic events. Further, TSPY copy number was found to vary significantly among tissues as well as among the same tissue type from different animals in a wide range from 7 to 224% of the calibrator. Our study shows significant autosomal and Y-chromosomal de novo somatic CNV in bulls.

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A novel approach to sexing bovine blastocysts using male-specific gene expression

Cited by 25 publications

References 54 publications

Enrichment of Y‐Chromosome‐Bearing Bull Spermatozoa by Swim‐up Through a Column

Enrichment of Y‐Chromosome‐Bearing Bull Spermatozoa by Swim‐up Through a Column

Initiation of X Chromosome Inactivation during Bovine Embryo Development

Somatic Mosaicism in Bulls Estimated from Genome-Wide CNV Array and <b><i>TSPY</i></b> Gene Copy Numbers

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