Sexing single bovine  blastomeres using TSPY gene amplification.
The  testis-specific protein Y-encoded gene (TSPY) is a Y-specific gene  present in variable copy number in many mammalian species, including cattle. We  tested the applicability of the TSPY gene as a Y-specific marker to  predict preimplantation embryo sex in Nelore (Bos indicus) cattle. Two  blastomeres were removed from each embryo. A total of 36 single blastomeres and  the remaining cells of their 18 matched in vitro conceived embryos were  screened for TSPY amplification by nested-PCR. The results obtained from  a single blastomere and the remaining cells of the same embryo were concordant  in all cases. All blastomeres (16/16) from eight embryos produced with sexed  sperm (specific for production of male embryos) were TSPY-positive. We  conclude that TSPY is a good male-specific marker, the usefulness of  which is probably enhanced by the high copy number. Other methods that are less  time-consuming, such as real-time PCR, could be improved with the use of the TSPY gene sequences to generate primers and/or probes. This is the first report  to demonstrate the applicability of the TSPY gene for sexing single  cells in cattle.
Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil.
M C A
MC
Carneiro
P L
PL
Takeuchi
A
A
Araújo
R B
RB
Lôbo
F P
FP
Elias
R A
RA
Vila
C L
CL
Miranda-Furtado
E S
ES
Ramos
D000818:Animals; D001757:Blastomeres / Q000378:metabolism*; D002417:Cattle; D018797:Cell Cycle Proteins / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004622:Embryo, Mammalian / Q000378:metabolism; D005260:Female; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012732:Sex Determination Analysis / Q000379:methods*
D018797:Cell Cycle Proteins