Publications (2)3.93 Total impact
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Article: Use of F1 progeny of HolsteinxZebu cross cattle as oocyte donors for in vitro embryo production and gene microinjection.
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ABSTRACT: This study was designed to determine the possibility of using F1 crossbreed cattle (HolsteinxZebu) as donors of oocytes for in vitro fertilization (IVF) and for pronuclear gene microinjection into in vitro-produced embryos. In the first part of the experiment oocytes from Bos taurus (Holstein), Bos indicus (Zebu) and F1 crossbred Bos taurusxBos indicus (HolsteinxZebu) genotypes were inseminated with Bos taurus (Holstein) semen and were allocated for in vitro embryo production using conventional IVF procedures. No differences were observed on the in vitro maturation (IVM) rates between breeds (HolsteinxHolstein:85%, ZebuxHolstein:84% and ZebuxHolsteinxHolstein:88%). Holstein cows yielded the highest number of cumulus oocyte complexes (6.8 per ovary) for in vitro maturation, differing (P<0.05) from ZebuxHolstein and ZebuxHolsteinxHolstein F1 by 5.1 and 5.8, respectively. However, the Holstein breed also yielded the lowest percentage of cleavage (45.1 vs 71.9% for ZebuxHolstein and 65.1% for ZebuxHolsteinxHolstein). Of the 3 genotypes, the hybrid F1 breed was the most efficient source of oocytes for the production of embryos capable of reaching morulae and blastocyst stages (76/250; P<0.001). In the second part of the study, 599 oocytes from the F1 breed were fertilized in vitro, 1 group of 150 oocytes was used for the determination of the optimal pronuclear visualization period. The highest number of oocytes with 2 pronuclei was observed between 24 to 28 h after IVF (27 to 42%). The remaining 399 oocytes were microinjected with a gene construct bearing the bacterial lacZ gene as the reporter for gene expression. Survival of embryos to microinjection was 73.8%, and 45.5% of them (50/110) cleaved in culture. Of the microinjected embryos, 1 out of 50 showed beta-galactosidase activity. These findings indicate that a tropical crossbreed of cattle (ZebuxHolsteinxHolstein) can be used as a source of oocytes for IVF programs and gene microinjection studies.Theriogenology 11/1994; 42(6):977-85. · 1.96 Impact Factor -
Article: Production of transgenic mice and rabbits that carry and express the human tissue plasminogen activator cDNA under the control of a bovine alpha S1 casein promoter.
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ABSTRACT: One-cell embryos from mice and rabbits were microinjected with a hybrid gene composed of 1.6 kilobases (kb) promoter/regulatory sequences of the bovine alphaS1 casein gene fused to the complementary DNA (cDNA) encoding for the human tissue plasminogen activator (htPA) and 3'untranslated sequences from rabbit beta-globin and SV 40 genes. Transgenic mice and rabbits that carry the htPA gene were obtained. In mice, 11 founder females were generated, and 6 of them expressed low levels (about 50 mug/ml) of htPA in their milk. Some of the transgenic mice showed rearrangements of the microinjected DNA sequences as judged by Southern blot analysis. A position-dependent expression of the transgene is suspected to occur. The only live-born founder transgenic rabbit obtained was a male, and it transmitted the transgene in a Mendelian fashion to F1 females, which expressed htPA at very low levels (8 to 50 ng/ml). Although the 1.6-kb bovine alphaS1 casein promoter that was used directs the synthesis of htPA specifically to the mammary gland, it may not be sufficient for a high level of expression.Theriogenology 06/1993; 39(5):1173-85. · 1.96 Impact Factor
