Successful suppression of endogenous α-1,3-galactosyltransferase expression by RNA interference in pig embryos generated in vitro     

Chi H  Shinohara M  Yokomine T  Sato M  Takao S  Yoshida M  Miyoshi K 《The Journal of reproduction and development》2012,58(1):69-76

RNA interference (RNAi) technology using small interfering RNAs (siRNA) has been widely used as a powerful tool to knock down gene expression in various organisms. In pig preimplantation embryos, no attempt to suppress the target gene expression with such technology has been made. The purpose of this study is to demonstrate that the RNAi technology is useful for suppression of endogenous target gene expression at an early stage of development in pigs. Alpha-1,3-Galactosyltransferase (α-GalT) is an enzyme that creates the Galα1-3Gal (α-Gal) epitope on the cell surface in some mammalian species, and removal of the epitope is considered to be a prerequisite for pig-to-human xenotransplantation. We decided to suppress the endogenous α-GalT mRNA expression in pig early embryos, since reduction of α-GalT synthesis is easily monitored by cytochemical staining with Bandeiraea simplicifolia isolectin-B(4), a lectin that specifically binds to the α-Gal epitope, and by RT-PCR analysis. Cytoplasmic microinjection of double-stranded RNA and pronuclear injection of an siRNA expression vector into the embryos generated in vitro resulted in a significant reduction in expression of the α-GalT gene and α-Gal epitope in blastocysts, at which stage the α-Gal epitope is abundantly expressed. Somatic cell nuclear transfer of embryonic fibroblasts stably transfected with an siRNA expression vector also led to a significant reduction in the level of α-GalT mRNA synthesis together with decreased amounts of the α-Gal epitope at the blastocyst stage. These results indicate that the RNAi technology is useful for efficient suppression of a target gene expression during embryogenesis in pigs and suggest the possibility of production of siRNA-expressing pigs for use in xenotransplantation.  相似文献   

Effects of force‐feeding and dietary cereals on gastrointestinal size,intestinal absorptive ability and endogenous nitrogen in ducks     

Z.‐X. Zhou  Y. Isshiki  K. Yamauchi  Y. Nakahiro 《British poultry science》1990,31(2):307-317

1. Gastrointestinal size, intestinal absorption of glucose and L‐methionine, and intestinal endogenous nitrogen in Pekin ducks were measured after the birds were subjected to force‐feeding with excessive amounts of a diet based on maize or barley for 12 d.

2. Force‐feeding significantly increased the weight and area of the crop, the weight of the proventriculus, the length and area of the jejunum and ileum, but decreased the weight of the gizzard and the thickness of its caudodorsal thick muscle. Feeding a barley‐based diet also decreased the thickness of caudodorsal thick muscle of gizzard.

3. Force‐feeding enhanced the absorption rates (μmol/cm²) of glucose and L‐methionine in both jejunum and ileum. Ducks fed on a maize‐based diet showed a slightly higher absorption rate of glucose compared to those fed on a barley‐based diet.

4. Force‐feeding caused a significant increase of endogenous non‐protein nitrogen (μg/cm²/h) in both jejunum and ileum and a slight increase of endogenous protein nitrogen in the jejunum.  相似文献   

Variation in levels of non-starch polysaccharides and endogenous endo-1,4-β-xylanases affects the nutritive value of wheat for poultry     

V. Cardoso  E. A. Fernandes  H. M. M. Santos  B. Maçãs  M. M. Lordelo  Luis Telo da Gama 《British poultry science》2018,59(2):218-226

1. Endo-1,4-β-xylanase is known to improve the nutritive value of wheat-based diets for poultry by degrading dietary arabinoxylans. However, broilers’ response to supplementation of wheat-based diets with exogenous endo-1,4-β-xylanase is not always observed.

2. In this study, 108 different wheat lots were analysed for levels of extract viscosity as well as for endogenous endo-1,4-β-xylanase activity, and the impact of these two variables in animal performance was tested.

3. Results revealed that endogenous endo-1,4-β-xylanase activity and extract viscosity content varied widely among different wheat lots. Thus, a trial was conducted to evaluate the efficacy of exogenous enzyme supplementation in broiler diets using wheats with different levels of extract viscosity and endogenous endo-1,4-β-xylanase activity.

4. The data revealed that exogenous enzyme supplementation was only effective when the wheat present in the diet had high levels of extract viscosity (14.8 cP) with low endogenous endo-1,4-β-xylanase activity (347.0 U/kg). Nevertheless, it is apparent that exogenous microbial xylanases reduce digesta extract viscosity and feed conversion ratio independently of the endogenous properties presented by different wheat lots.

5. The data suggest that extract viscosity and/or endogenous endo-1,4-β-xylanase activity affect the response to enzyme supplementation by poultry fed on wheat-based diets.  相似文献