Absence of Fetal Cells in Bovine Jugular and Uterine Vein Blood at a Level of 1 in 10,000.

Kadokawa, Hiroya; Takusari, Naozumi; Minezawa, Mitsuru; Takahashi, Hitomi; Kariya, Takayoshi

doi:10.1262/jrd.42.205

Cited by 6 publications

(3 citation statements)

References 15 publications

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“…However, since humans have a hemochorial type of placenta and cows have a (syn) epitheliochorial type and the bovine placenta is anticipated to prevent transplacental cell leakage into the blood of the cow, it is assumed that fetal cells could not be used for prenatal sexing in cattle because they are very rare or may be absent in bovine maternal blood [7]. However, there was a report of successful prediction of fetal sex through amplification of the SRY gene from the blood of pregnant cows [8].…”

Section: Discussionmentioning

confidence: 99%

“…Although it was recently demonstrated that fDNA in women and rhesus monkeys could be used as a tool for fetal sex determination as early as at the 5th gestational week using the Y chromosome DNA sequence as a probe for screening male fetuses with an accuracy rate of 100% [3][4][5][6], little is known about fDNA in domestic animals. In 1996, Kadokawa et al believed, based on their previous research, that fetal cells were very rare or may be absent in bovine maternal blood and that they could not be used for prenatal sexing by a PCR method [7]. However, Xi et al reported in 2006 that they had successfully predicted fetal sex by amplifying the SRY gene from pregnant cow blood [8].…”

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Prediction of Fetal Sex by Amplification of Fetal DNA Present in Cow Plasma

Wang

Cui

Cheng

et al. 2010

Journal of Reproduction and Development

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Abstract. The aim of the present study was to predict fetal sex at different time points of gestation in cattle by detecting the fetal SRY gene in cow plasma. Plasma DNA was extracted from the blood samples of 110 pregnant cows during the gestational period of 30 to 242 days. Nested PCR was employed to detect the fetal SRY, which the male fetus carries exclusively, in cow plasma. The cows positive for SRY were predicted to carry male fetuses. The results showed that the fetal DNA from cow plasma was successfully amplified and that fetuses could be sexed with an overall accuracy rate of 100% (43/43) for males and 91.0% (61/67) for females and with accuracy rates of 100% (3/3) for males and 85.7% (12/14) for females at 30 EN 59 days of gestation and 100% (40/40) for males and 92.5% (49/53) for females at more than 2 months of gestation, respectively. This suggests that the molecular method developed here could be used in sex prediction for fetuses. Key words: Cow plasma, Fetal Sex, Nested PCR, Sry (J. Reprod. Dev. 56: [639][640][641][642] 2010) he discovery of fetal DNA (fDNA) in maternal circulation [1] and transcervical cells has led to noninvasive prenatal genetic diagnosis and sex prediction for fetuses during early pregnancy in women [2]. Although it was recently demonstrated that fDNA in women and rhesus monkeys could be used as a tool for fetal sex determination as early as at the 5th gestational week using the Y chromosome DNA sequence as a probe for screening male fetuses with an accuracy rate of 100% [3][4][5][6], little is known about fDNA in domestic animals. In 1996, Kadokawa et al. believed, based on their previous research, that fetal cells were very rare or may be absent in bovine maternal blood and that they could not be used for prenatal sexing by a PCR method [7]. However, Xi et al. reported in 2006 that they had successfully predicted fetal sex by amplifying the SRY gene from pregnant cow blood [8]. However, only 30 cows were tested in their report, and the overall accuracy rate was 80%; the accuracy rate for 30-59 days of gestation was 60% (6/10), and they also did not analyze the sequences of the PCR products.The aim of present study was to verify the possibility of detecting fDNA in cow plasma and develop a noninvasive method of fetal sex identification in pregnant cows in order to offer a way for sex selection in dairy production and for early selection of cattle in breeding programs as well as for elimination of harmful genes in bulls in the fetal phase. Materials and Methods Blood samples and DNA isolationTen ml of blood was collected from each of 110 pregnant Holstein cows that were 3-6 years of age and at 30-242 days of gestation through the vena caudalis by single-use syringe. All the blood samples were put into acid citrate dextrose anticoagulant tubes and centrifuged at 4 C and 1600 × g for 10 min. Supernatants were stored at -20 C for further analysis. Blood samples from 2 virgin heifers at 3 months of age with no history of contact with bulls or semen and 6 bulls that were 5 yea...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Prediction of Fetal Sex by Amplification of Fetal DNA Present in Cow Plasma

Wang

Cui

Cheng

et al. 2010

Journal of Reproduction and Development

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“…Apesar da invasão trofoblástica em bovinos ser considerada restrita ao epitélio uterino, e a possibilidade do microquimerismo fetal em bovinos ter sido descartada por Kadokawa et al (1996), ele foi originalmente descrito por Xi et al (2006) (ROSBOTTOM et al, 2008).…”

Section: Microquimerismo Em Bovinosunclassified

Ocorrência e mecanismos do microquimerismo fetal em gestações bovinas

Barreto¹

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RESUMO BARRETO, R. S. N. Ocorrência e mecanismos do microquimerismo fetal em gestações bovinas [Occurrence and mechanisms of fetal microchimerism in bovines pregnancies]. 2011. 68f. Dissertação (Mestrado em Ciências) -Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, 2011. O sucesso da gestação depende da adequada comunicação materno-fetal, que em algumas espécies têm um contato mais íntimo devido à capacidade migratória de populações de células trofoblásticas. Nos bovinos esse mecanismo é realizado pelas células trofoblásticas gigantes (CTGs), com invasão limitada até a lâmina basal do epitélio materno. Apesar dessa leve invasão das CTGs, é possível encontrar células fetais circulantes no sangue periférico da vaca gestante, levando ao microquimerismo fetal. Além de toda uma sinalização local e sistêmica e mudanças conformacionais, a migração das CTGs também é dependente da tolerância imunológica do epitélio materno que possui uma baixa expressão de MHC de classe I. Em contrapartida, o trofoblasto expressa MHC de classe Ib para impedir a ativação das células natural killers uterinas (uNK) contra ele mesmo. Neste contexto, o objetivo desse trabalho foi estudar a ocorrência e contribuir para o entendimento dos mecanismos da migração celular na placenta bovina, com marcadores exclusivos do cromossomo Y e de um modelo de clone transgênico expressando a proteína GFP. A hipótese testada foi que o microquimerismo fetal observado mediante a detecção do gene TSPY no sangue periférico da vaca gestante de embrião macho, e de GFP nos tecidos placentários maternos, associado à expressão de MHC classe 1b (Qa2) na interface materno-fetal. Para tanto, 153 embriões produzidos por fertilização in vitro (FIV) foram transferidos, resultando em 34 embriões machos e 31 fêmeas no dia 62 de gestação, quando foi realizada a coleta de sangue periférico da receptora. Dentre estas gestações, foram selecionadas de 25 machos, 4 fêmeas e 5 perdas gestacionais (confirmadas no D39 por ultrassonografia) para detecção de TSPY. Também foram produzidas gestações de clones transgênicos, expressando GFP com 30, 60 e 90 dias que foram utilizadas para a detecção de mRNA e a proteína GFP. Nas gestações de FIV 60% dos embriões machos, 50% das fêmeas e 40% das perdas gestacionais foram positivos para TSPY. A detecção de TSPY nas gestações de fêmeas possivelmente é resultante da persistência do microquimerismo de gestações anteriores. Nas gestações de clones transgênicos, observou-se a presença de mRNA e proteína GFP no endométrio, também indicando migração nesta região ou o transporte da GFP, e outros conteúdos do trofoblasto, para o epitélio materno. Nos placentônios, usando anticorpo anti-GFP pode-se ver a marcação positiva tanto no trofoblasto como no epitélio materno, possivelmente decorrente de liberação das CTGs no estroma endometrial após a fusão. As CTGs, quando em formação sincicial, têm a sua expressão de GFP diminuída, o que também foi observado, utilizando-se anticorpo anti-Qa2 (antígeno murino para MHC classe...

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Application of polymerase chain reaction for fetal gender determination using cervical mucous secretions in the cow

Divar¹,

Sharifiyazdi²,

Kafi³

2012

Vet Res Commun

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In the current study we aimed to use PCR to investigate the presence of fetal DNA in the bovine (Bos taurus) cervical secretions and maternal serum, and to assess the effectiveness of this method in fetal gender determination. Pregnant uteri and pre-slaughter maternal blood samples were collected from 21 Holstein Frisian cows in a local abattoir. Overall, 13 male and 8 female fetuses were included in the study. Cervical mucus was sampled at the laboratory. After DNA extraction, the PCR amplified a 280 bp fragment from the X-chromosome and a 217 bp fragment from the Y-chromosome based on a sex-related polymorphism in the amelogenin locus. The presence of fetal Y-chromosome was confirmed in seven out of 13 cervical mucus samples collected from cows with male fetuses. Overall test sensitivity for correct sex determination based on PCR assay on cervical samples was equal to 71.4 ± 2 %. In contrast, no fetal Y-chromosome DNA was detected in maternal serum samples from cows with male fetuses. This is the first report on validating the presence of fetal DNA material in the bovine cervical mucus and its potential usefulness for fetal sexing. Further investigations are needed to maximize the accuracy and evaluate the practicality of this approach.

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Absence of Fetal Cells in Bovine Jugular and Uterine Vein Blood at a Level of 1 in 10,000.

Cited by 6 publications

References 15 publications

Prediction of Fetal Sex by Amplification of Fetal DNA Present in Cow Plasma

Prediction of Fetal Sex by Amplification of Fetal DNA Present in Cow Plasma

Ocorrência e mecanismos do microquimerismo fetal em gestações bovinas

Application of polymerase chain reaction for fetal gender determination using cervical mucous secretions in the cow

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