Sex of progeny from bovine embryos sexed with a rapid Y-chromosome-detection assay

Sex Determination by PCR in Single Bovine Blastomeres Biopsied by Extrusion Method at the 8-cell Stage.

“…Nevertheless, because a success rate of less than 100% in cattle was mentioned in many reports [3,5,7], it is difficult to obtain 100% in cattle, unlike mice, for which 100% was reported [9]. In this study, determined sex was inconsistent among 1/8, 2/8, and 4/8 derived from the same embryo, and between 1/ 8 and 7/8 from the same embryo.…”

“…Among these methods, the method using a male-specific antigen and the method using sex difference in the rate of development are not always accurate, and the overall sexing rates of the method for measuring enzymes linked to the X chromosome and the method for detecting the Y chromosome are low. Therefore, the method for detecting Y chromosome-specific nucleotide sequences by amplifying the sequence by polymerase chain reaction (PCR method) is considered to be promising, and has been developed for cattle [3][4][5][6][7], pigs [8] and mice [9,10]. As an advantage of the PCR method, tests can be rapidly and accurately performed on only a few cells, and commercial use in cattle has begun, but with the current biopsy method, blastomeres are collected by excising mural trophectoderm cells with the zona pellucida at the blastocyst stage, and there is a risk of affecting the normal development of the fetus.…”

Fractionation of X and Y Chromosome-Bearing Bovine Spermatozoa through Sugar Gradients for Sex Predetermination in Dairy Cattle

“…It can be achieved either by sexing of early embryos or by separation of X and Y chromosome-bearing spermatozoa. Sexing of embryos is practiced in both humans and livestock, especially cattle, by performing the DNA polymerase chain reaction or in situ hybridization to detect Y specific DNA, in cells taken from the early embryo [1], [2]. Despite the success of embryo sexing, sperm separation is a more preferred method for sex preselection, because it would avoid the need for embryo manipulation and exclusion.…”

Conception rate after sex determination and cryopreservation of D7 bovine embryos:

“…Employed methods include cytogenetic analysis (Singh and Hare, 1980;Rall and Leibo, 1987), immunological assays (White et al, 1982;Booman et al, 1989), detection of metabolic differences between male and female embryos (Williams, 1986;Monk and Handyside, 1988), analysis of chromatin with Y-specific DNA probes (Leonhard et al, 1987;Bondioli et al, 1989;Kobayashi et al, 1998) and analysis on the base of differences in cleavage rates (Avery et al, 1989). Better results were published later using PCR amplification of specific DNA sequences to determine embryonic sex in cattle (Herr et al, 1990;Schroder et al, 1990;Peura et al, 1991), pigs (Pomp et al, 1995), horses (Peippo et al, 1995), humans (Handyside et al, 1990) and mice (Han et al, 1993). Recently published results support the suitability of the PCR method for sex determination in cattle on the basis of its high accuracy and quick results (Thibier and Nibart, 1995;Lopes et al, 2001;Ekici et al, 2006;Yu et al, 2007).…”

“…Embryo sexing was performed with a commercial polymerase chain reaction (PCR) kit using primers specific for the Y-chromosome determinant (YCD) according to the manufacturer's instructions (Herr et al, 1995). The PCR product was detected by UV light in agarose gel with ethidium bromide (10 mg/ml in distilled water) and the embryos were scored as Y-chromosome determinant positive (male) or Y-chromosome determinant negative (female), respectively.…”