Length difference between equine ZFX and ZFY genes and its application for molecular sex determination

Abstract

Purpose We analyzed the sex chromosome-encoding ZFX-ZFY genes and tested molecular sexing using the amplification patterns of intron 9 of ZFX-ZFY in the horse. Methods and resultsThe amplification of the ZFX-ZFY produced two distinct patterns, reflecting sexual dimorphism based on a length difference between the X and Y chromosomes. The amplification products from foals showed two distinct bands: one was common to all foals and mares, indicating that this band was amplified from ZFX, while the other was specific to some foals, indicating that it was from ZFY. The result based on the PCR assay was identical to the results of amplification of the Y chromosome-specific SRY gene and those of investigations of the phenotypic gender in three different horse populations. Conclusion We suggest that this PCR strategy for determining sexes by comparing the amplification patterns of ZFX-ZFY genes is a convenient and precise method for discriminating sexes in horses.Most mammals have an X-Y chromosome sex determination system. Sexual differentiation is determined primarily by the presence or absence of the Y chromosome, and the sex determining region Y (SRY) gene plays a key role in inducing male development, while inhibiting female differentiation [1][2][3].Control of the sex ratio of animals is desirable in the livestock industry. In mammals, the X-Y homologous sequences for the X-and Y-linked genes zinc finger, amelogenin-X and -Y (AMELX-AMELY), and SMCX/SMCY have been compared [4][5][6]. PCR assays for molecular sexing are often more useful because they provide sensitive, precise, rapid, and reliable results. To date, PCR-based sex typing has been used for embryonic sexing in cattle, mice, and pigs [7][8][9][10][11][12]. To establish a rapid and precise molecular sexing method using PCR amplification for horse breeding, we analyzed the sequence characteristics and sexual dimorphism of the ZFX and ZFY genes and compared the results to those obtained from Y chromosome-specific SRY gene amplification and phenotypic investigation.Total of 128 genomic DNA samples were collected from three horse populations (61 Thoroughbreds, 20 Tsushima native horses from Japan, and 47 Jeju native horses from South Korea). To amplify the intron 9 of ZFX and ZFY genes, the primer pair was designed from exon 8 and 10 of the sequences of human and cattle [13]. The primer Capsule Length difference was found in PCR amplification products between equine ZFX and ZFY genes. This sexual dimorphism may be a diagnostic molecular marker for determining the sexes in the horse.

Sex chromosomal abnormalities associated with equine infertility: validation of a simple molecular screening tool in the Purebred Spanish Horse

“…) or zinc finger protein genes (Han et al . ), as there is no discordance between genotypic and phenotypic sex. Therefore, the number of undiagnosed cases could be higher and the prevalence underestimated.…”

Sex Determination Using RNA-Sequencing Analyses in Early Prenatal Pig Development

“…The control of animals sex ratio is desirable in livestock production [20], since the expression of genes that affect a wide range of economically important traits may be influenced by sexual dimorphism [12]. Considering that the sex determination is equivalent to testis determination [30], the testis could be easily used to determine male and female individual rates.…”

“…Several studies have demonstrated how to identify the conceptuses sex at early stages of development using molecular techniques, as PCR (polymerase chain reaction) from Y chromosome-linked genes [13,[18][19][20]. The PCR provides sensitive, precise, rapid, and reliable results [20]. However, if a RNA-seq study has already been performed on conceptuses to obtain a broader knowledge of the transcriptome under experimental conditions (e.g.…”

Disorder of Sexual Development in a Mare with an Unusual Tentative Mosaic Karyotype: 63,X/64,Xdel(Y)

“…High molecular weight genomic DNA was extracted by a proteinase K/ phenol extraction method. PCR amplification of the SRY gene from genomic DNA was performed using a primer pair described in Han et al [2010], which results in a 714-bp amplification product. Additional PCR primers for the equine Y chromosome (ECAY) contigs [Paria et al, 2011] were used: YM2 (contig I, 119 bp) , YE1 (contig II, 199 bp) , NLGN4Y (contig III, 156 bp) [Paria et al, 2011], AMELY (contig IV, 160 bp) [Hasegawa et al, 2000], ZFY (contig V, 342 bp) [Lindgren et al, 2001].…”