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Efficient production of pronuclear embryos in breeding and nonbreeding season for generating transgenic sheep overexpressing TLR4.

Li Y, Lian D, Deng S, Zhang X, Zhang J, Li W, Bai H, Wang Z, Wu H, Fu J, Han H, Feng J, Liu G, Lian L, Lian Z - J Anim Sci Biotechnol (2016)

Bottom Line: Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons (P > 0.05).The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.The over-expression of TLR4 had no adverse effect on the growth of the sheep.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing, 100193 China.

ABSTRACT

Background: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons (breeding and non-breeding season) on superovulation and the imported exogenous gene on growth.

Results: In total of 43 donor ewes and 166 recipient ewes in breeding season, 37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep. Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons (P > 0.05). The positive rate of exogenous TLR4 tested were 21.21 % and 22.58 % in breeding and non-breeding season by Southern blot. The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group (P < 0.05). The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.

Conclusions: Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year. The over-expression of TLR4 had no adverse effect on the growth of the sheep.

No MeSH data available.


Related in: MedlinePlus

The effects of different seasons on superovulation of donors. a No. of embryos recovered. b No. of left and right oviduct embryos recovered. c Fertilization rate. BS = Breeding Season, N = 43. NBS = Non-breeding Season, N = 37
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Fig1: The effects of different seasons on superovulation of donors. a No. of embryos recovered. b No. of left and right oviduct embryos recovered. c Fertilization rate. BS = Breeding Season, N = 43. NBS = Non-breeding Season, N = 37

Mentions: In the breeding season, 515 zygotes and 3 unfertilized ova were flushed out from 43 donor ewes. In the non-breeding season, 552 zygotes and 5 unfertilized ova were flushed out from 37 donor ewes. There was no significant difference of no. of embryos recovered (12.05 ± 5.22 versus 15.05 ± 6.22, P > 0.05, Fig. 1a) between sheep from breeding season and non-breeding season. Subsequently, we further compared the difference of ovulation in left and right ovary, and the results showed that there were no significant differences in both left and right oviduct between breeding and non-breeding season (Fig. 1b), as the fertilization rate (99.48 ± 2.84 % versus 99.29 ± 1.83 %, P < 0.05, Fig. 1c).Fig. 1


Efficient production of pronuclear embryos in breeding and nonbreeding season for generating transgenic sheep overexpressing TLR4.

Li Y, Lian D, Deng S, Zhang X, Zhang J, Li W, Bai H, Wang Z, Wu H, Fu J, Han H, Feng J, Liu G, Lian L, Lian Z - J Anim Sci Biotechnol (2016)

The effects of different seasons on superovulation of donors. a No. of embryos recovered. b No. of left and right oviduct embryos recovered. c Fertilization rate. BS = Breeding Season, N = 43. NBS = Non-breeding Season, N = 37
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4940989&req=5

Fig1: The effects of different seasons on superovulation of donors. a No. of embryos recovered. b No. of left and right oviduct embryos recovered. c Fertilization rate. BS = Breeding Season, N = 43. NBS = Non-breeding Season, N = 37
Mentions: In the breeding season, 515 zygotes and 3 unfertilized ova were flushed out from 43 donor ewes. In the non-breeding season, 552 zygotes and 5 unfertilized ova were flushed out from 37 donor ewes. There was no significant difference of no. of embryos recovered (12.05 ± 5.22 versus 15.05 ± 6.22, P > 0.05, Fig. 1a) between sheep from breeding season and non-breeding season. Subsequently, we further compared the difference of ovulation in left and right ovary, and the results showed that there were no significant differences in both left and right oviduct between breeding and non-breeding season (Fig. 1b), as the fertilization rate (99.48 ± 2.84 % versus 99.29 ± 1.83 %, P < 0.05, Fig. 1c).Fig. 1

Bottom Line: Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons (P > 0.05).The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.The over-expression of TLR4 had no adverse effect on the growth of the sheep.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing, 100193 China.

ABSTRACT

Background: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons (breeding and non-breeding season) on superovulation and the imported exogenous gene on growth.

Results: In total of 43 donor ewes and 166 recipient ewes in breeding season, 37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep. Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons (P > 0.05). The positive rate of exogenous TLR4 tested were 21.21 % and 22.58 % in breeding and non-breeding season by Southern blot. The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group (P < 0.05). The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.

Conclusions: Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year. The over-expression of TLR4 had no adverse effect on the growth of the sheep.

No MeSH data available.


Related in: MedlinePlus