Method of enhancing engraftment of embryos by transplantirovannam previously inseminated ewes - recipients
(57) Abstract:The invention relates to agriculture - sheep breeding method embryo transfer securities genotype. The method is carried out transplantation (transplantation) of zygotes in the non-pregnant uterine horn 5 days after pre-insemination of ewes recipient. The method provides increased engraftment of embryos in ewes recipient after transplantation. The invention relates to the field of agriculture, particularly sheep, for sheep rearing method embryo transfer securities genotype.The closest analogue, selected as a prototype, is a method for twinning by transplanting embryos in each uterine horn of a cow or embryo transfer in the contralateral horn of the uterus in relation to the yellow body of the ovary previously inseminated recipients /Ernst L. K., Sergeev, N. And. "Transplantation of embryos of farm animals", 1989, 173-175 C./.The method of obtaining twins by transplanting embryos cows-recipients is that involves transplantation of one embryo in each horn of the uterus of the recipient or transplant one Embry is and. For successful engraftment of embryos is important not only for their usefulness, but also the physiological state of the recipient, in particular morpho-functional status of the endometrium horns of the uterus. Also the determining factor of engraftment embryo recipients are the overall development of the embryo from the donor and differentiation of the endometrium in the recipient. The successful settling down transplanted embryos in sheep depends on the stage of their development and synchronization of the estrous cycle in the recipient with the stage and age of embryos, number of embryos transferred into the uterine horns and the number of yellow bodies in the ovaries of the recipient. When the gap of the age of the embryos from the astral cycle of the recipient on 1 day settling down is 72%, for 2 days - 70%, for 3 days - 7%. When the advancing age of the embryo astral cycle of the recipient on 1 day settling down is 65%, for 2 days - 50% and for 3 days - 4%.The purpose of the invention is improving the engraftment of embryos in ewes recipient after transplantation.The objective is achieved by the fact that the transplanted embryo in the contralateral /pregnant/ horn of the uterus in relation to the yellow body of the ovary 5 days after pre-insemination of ewes recipient as the body. As a result of these hormonal shifts during the establishment of pregnancy there are changes in secretory activity of the uterus and in the structure of its mucous membrane, which play a crucial role in the development and engraftment of the embryo. When fruitful preliminary insemination of ewes recipient is provided corresponding optimal level of progesterone in the blood of these animals, characteristic for phase active luteum during pregnancy and the transplanted embryos normally engraftment, as they are already prepared in optimal conditions uterine environment formed by the embryo after the preliminary fruitful insemination recipients.Comparative analysis of the proposed solutions with the prototype shows that transplantation of embryos to the previously inseminated ewes-recipients is to enhance the engraftment them at the expense created ready-optimal conditions of the uterine environment of the recipient zygote, after developing the preliminary effective insemination. The zygote develops in the uterine environment after pre-insemination of ewes recipient submits to the receptors of the uterus signals the beginning of suggestion, under the action of which occur proliferation of the endometrium of the uterus and the increase in the content of glycogen, enzymes and other biologically active substances, i.e., formed ambitrol necessary for the further development of the embryo before the establishment of the placental connection with the parent body.Comparative analysis of the proposed solutions are not found close to the prototype and therefore meets the criterion of "novelty". The analysis of the hypothesis of implantation of embryos in the non-pregnant uterine horn previously inseminated recipients allows to make a conclusion about the absence of these symptoms that are similar to the essential distinguishing features of the claimed method of enhancing engraftment of embryos by transplantirovannam previously inseminated ewes - recipients and, therefore, provides the claimed solution according to the criterion of "significant differences".Thus, the use of the proposed method of enhancing engraftment of embryos by transplantirovannam previously inseminated ewes-recipients provides an optimal uterine environment for implantable embryo prepared by the zygote, formed and a developing after a fruitful tentative is essof, occurred in the body and reproductive organs of recipients in the period of oestrus and establishment of pregnancy after a fruitful breeding ewes-recipients, promotes optimal engraftment of transplanted embryos us. Method of enhancing engraftment of embryos by transplantirovannam previously inseminated ewes-recipients, ensuring optimal intrauterine environment for the transplanted embryo, developing zygote after preliminary fruitful insemination recipients, characterized in that on the 5th day after insemination of ewes-recipients transplanted (transplanted) in their non-pregnant horn of the uterus the embryo securities genotype and thereby implantable zygote gets prepared in optimal conditions of the uterine environment of the recipient, which contributes to the successful nidali and implantation of an embryo.
SUBSTANCE: method involves using product containing spermatozoa treated by drying with freezing to humidity level of 1% and having injured membrane or spermatozoon head so that spermatozoon nucleus retains its genetic validity enough for fertilization. The spermatozoon heads fertilize an isolated oocyte after rehydration and microinjection being done. The retained genetic integrity is enough for fertilizing an oocyte and producing living descendants. Method involves collecting living mature spermatozoa, making spermatozoa suspension in special purpose physiological medium, freezing the spermatozoa suspension for producing frozen spermatozoa, drying the frozen spermatozoa or spermatozoa heads in vacuum to humidity level of 1%, making rehydration of spermatozoa or spermatozoa heads with injured membrane and selecting those retaining nucleus of genetic integrity. The selected spermatozoa or spermatozoa heads are used for fertilizing isolated oocytes with living descendants being produced.
EFFECT: enhanced effectiveness in producing living descendants.
39 cl, 5 dwg, 1 tbl