For a large sclase production of genetically identical or cloned animals, the effect of cryopreservation by vitrification on the post-thaw viability of nuclear transplant rabbit embryos were investigated. The embryos of 16-cell stage were collected from the mated does at 48 hours post-hCG injection, and they were synchronized to G1 phase of 32-cell stage were injected into enucleated recipient cytoplasms by micromanipulation. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused and activated by electrical stimulation. After in vitro culture for 48h, the nuclear transplant embryos developed to morula stage were cryoperserved with EFS solution by vitrification method. The forzen nuclear transplant embryos were thawed and cultured for 72h and the nuclear transplant of blastomeres under a fluorescence microscopy. The in vitro development to blastocyst of intact-fresh and intact-frozen 16-cell embryos was found to be 96.9 and 63.9%, respectively. The in vitro development to blastocyst of nuclear transplant and frozen-thawed nuclear transplant embryos was found to be 74.5 and 42.9%, respectively. Also, their mean blastomere numbers and mean cell cycles/day was 153 and 105, 145 and 1.34, respectively. From the above results it was concluded that the present cryopreservation by vitrification of nuclear transplant rabbit embryos might be useful though was decreased significantly.
Different genetic parameters for weaning weight and wool traits were estimated using restricted maximum likelihood (REML) in Angora rabbits. Total wool yield of first (I), second (II) and third (III) clips were taken as a separate trait under study. The records from more than 2,700 animals were analysed through fitting six animal models with various combinations of direct and maternal effects. A log likelihood ratio test was used to select the most appropriate model for each trait. Direct heritability estimates for the wool traits were found to be moderate to high across different models. Heritability estimates obtained from the best model were 0.24, 0.22, 0.20 and 0.21 for weaning weight, clip I, II and III; respectively. Maternal effects especially due to permanent environment had higher importance at clip I and found to be declining in subsequent clips. The estimates of repeatability of doe effect on wool traits were 0.44, 0.26 and 0.18 for clip I, II and III; respectively. Weaning weight had moderately high genetic correlations with clip I (0.57) and II (0.45), but very low (0.11) with clip III. Results indicated that genetic improvement for wool yield in Angora rabbit is possible through direct selection. Further, weaning weight could be considered as desirable trait for earliest indirect selection for wool yield in view of its high genetic correlation with wool traits.
To the rabbit and guinea pig, tartrazine was administered, either parenterally through venous or peritoneal injection or orally, with the amount of 7.5-50 mg/kg body weight in case of rabbit and 1,000-2,000mg per animal in case of guinea pig, in order to observe acute toxic effect of the dye in the animals. Blood serum was analyzed chemically and all the internal organs were microscopically observed for this purpose. The results point to the fact that severe hemorrhagic and congestive findings in most of the organs can be induced by the amount of the dye utilized, and particularly liver damage could be resulted.
This experiment was carried out to improve a simple and effective procedure of egg transfer which is considered to be the most useful technique for the improvementand proliferation of domestic animals. Several experiment procedures such as superovulation, surgical recovery of ovulated egg, in vitro capacitation of ejaculated spermatozoa, in vitro fertilization and culture of embryo were conducted. The results obtained in this experiment were summarized as follows: 1. In rabbits treated with PMS in combination with estradiol and HCG, 10 to 43 eggs were obtained in one rabbit and the average ovulation number of 20 rabbits was 21 eggs. 2. Six to 24 eggs (average 13 eggs) were recovered from the removed oviducts by the methods of flushing technique and the average recovery rate of 20 rabbits was 61.9 percent. 3. Most rabbit spermatozoa ejaculated by artificial vagina were capacitated in vitro by the culture of the spermatozoa with PBI medium at 37$^{\circ}C$ for 4 hours after removal of seminal plasma. 4. Normal fetilization following in vitro fetilization was observed in 88 (42.7%) of 206 eggs. 5. The number of eggs developed to 2-, 4-, and 8-cell stage following in vitro culture with PBI medium at 37$^{\circ}C$ was 26 (70.3%), 20 (54.1%) and 17 (45.9%) of 37 eggs used for in vitro culture.
The influence of guanethidine on the renal function was investigated in the rabbit. Guanethidine, 1-10mg/kg, i.v., produced no marked change in the renal function, while second and successive doses of guanethidine elicited a significant increase in urine flow and electrolyte excretion as well as renal plasma flow and glomerular filtration rate. It was suggested that the diuretic action was brought about by improvement of hemodynamic state in the kidney ; increased filtration as a result of increased renal perfusion. Atropine alone did not significantly influence the renal function but pretreatment of animals with atropine, 4 mg/kg i.v., completely abolished the diuretic action of guanethidine. It is suggested that guanethidine influences the renal function by activating parasympathetic nervous system or some cholinergic mechanism in the kidney.
This stduy was carried out to investigate the effects of Isoimmunization by sperm and seminal plasma on density of immunoglobulins in reproductive tract fluids and serum of immunized rabbits. The results obtained were summarized as follows: 1. Antibody titers against sperm and seminal plasma antigen ranged from 8 to 64 to 512, respectively. 2. All immunoglobulins; IgG, IgA and IgM were detected with Indirect ELISA method in the uterine and oviductal fluids as well as the sera of immune rabbits. 3. Concentrations of IgGs in the uterine and oviductal fluids of rabbits immunized with sperm and seminal plasma were higher than those of the control rabbits, but not showed any differences in sera. 4. Amount of IgA in the sera and oviductal fluids of control animals was more than that of the immune animals, while that of IgA in the uterine fluids of control and seminal plasma-immunized animals was higher as compared to sperm immune animals. 5. Average concentration of IgM in the uterine fluids of control and seminal plasma-immunized rabbits was higher than that of sperm-immunized ones. In the oviductal fluids, average concentrations of IgM of immune rabbits was higher than that of immune rabbits.
In the previous studies, the present author found that high proportion of the follicular oocytes from mouse and rabbit ovaries are able to resume their maturation division in the anterior chamber of the eye in which they have been incubated by auto- or homoplastic transplantation. Especially in the case of the homoplastic transplantation, it was known that no trouble has been detected in the process of resumption of the oval maturation in particular connection with the antigen-antibody reaction between donor and recipient. These findings provide a possibility that the follicular oocytes from various animals would be matured in the eye even after the xenoplastic transplantation. Under such an assumption, the present studies were performed to examine the behavior of the follicular oocytes in the eye chamber of the animals of different species. For the donor of the follicular oocytes, domestic rabbits, albino rats of Sprague-Dowley strain, and albino mice of A-strain bred in our laboratory were used. The oocytes obtained from the ovarian follicules were introduced to the anterior chamber of the eye of different species of animals, with an exception of rabbit in which only the female animals were used as a recipient. The procedures of collection of ova, introduction to the eye, harvest from the eye ball, fixation, and staining were the same as mentioned in the previous reports (Cho, 1967b; Cho and Kim, 1968). The conclusions obtained are summarized as below. 1. The rabbit follicular oocytes are able to mature in the eye chambers of both male mouse and rat, although the proportion of the maturation is lower than when they are incubated autoplastically in the eye. When the ova were incubated in the male mouse eye for 24 hours, 21 per cent of them showed chromosomes at metaphase I and II, whereas the rate was 32 per cent when they were incubated in the eye of the male rat. These are apparently low comparing to the rate of 52 per cent of autoplastic transplantation. 2. When rat follicular oocytes were transferred into the mouse eye chamber and recovered after 24 hours, 43 per cent of them produced the mataphase I and II chromosomes. This proportion was higher than the result of the homoplastic transplantation which yielded 23 per cent of the ova on maturation. 3. The most striking result was found in the experiment with mouse follicular oocytes. Seventy-six per cent of the oocytes resumed their maturation division within 24 hours after they were transferred into the male rat eye chamber, and this figure was significantly high compared to the result o 55 per cent obtained by the homoplastic transplantation. In the rat eye, the induction of the degenerative ova also was low (19%). On the other hand, the proportion of the oval maturation decreased to 45 per cent, while that of degeneration increased 33 per cent when they were incubated in the eye of the female rabbit. 4. It was apparent from the present experiments that the follicular oocytes can reveal their activation to maturation in the eye chamber which contains aqueous humor which is known to be composed of low protein content and of very little gamma-globulin which acts as an antibody(Oser, 1965), and that it shows higher osmolarity than blood serum(Levene, 1958). Taking these properities into consideration the humor may provide unfavourable environment to the cells and tissues incubated in. However, it could be noteworthy finding that only the follicular oocytes in the eye of the different species can grow in healthy condition although the maturation rates are varied with the animal species. The fact that the rabbit follicular oocytes show the lower proportion in maturation may be due to the greater amount of the yolk granules in the egg cytoplasm than those in the mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat resumed their maturation process in greater proportion would e explained by the fact that the rat eye chamber particularly provides the better environment to the mouse oocytes than the eye chamber of mouse does.
SMZ is one of the most widely used antibacterial agents in veterinary medicine. It is also used as a growth promotant in many species of domestic animals There are marked species differences in its metabolism and pharmacokinetics. However, its pharmacokinetic and metabolism in rabbits. which are ragarded not only as good laboratorty animals hut also as good economical animals in its own, are lacking. Sex-differences in drug metabolism are well recognized in wide range of animal species including rats. Males are known to he more active than females. It is also know that there are Significant differences in the direction of metabolic pathways. But recently, female goats are reported to be more active in the metabolie capacity of SMZ than the other sex by Dutch researchers at Utrecht. Therefore, it is not easy to make general conclusicn of having higher SMZ metal-die capacity in the male compared to the opposite sex in every animal species. In this regard, the study on metabolic pattern of SMZ in rabbits, which are regarded as hervivorous, is of interest because the dietary habbits of rabbit are comparable to thai of goal, NEW Zealand White rabbits of each sex were given SMZ(35mg/kg) as a bolus injection into the marginalean, vein in order to study its pharmacokinetic profiles(using plasma) anc metabolic pattem(24h urine) as specified in the methods anc materials. 1. In the rabbit, the major metabolic pathway of SMZ was the acetylation(the formation of $N_4AcSMZ$). There were hydroxylation pathways(50HSMZ, $6CH_2OHSMZ$) as well, in the metabolism of SMZ in the rabbit, but minor pathways. 2. No sex differences in the metabolic direction of SMZ and its metabolites formation were found from the urinary excreted metabolites of SMZ out of 24h collected urine. 3. The concentration-time curves of SMZ(35mg/kg, iv) in the plasma compartment were fitted to a one-compartment open model when using a computer program(NONLIN). There was also no difference in the pharmacokinetic pattem of SMZ between two sexes. 4. The emergence of $N_4AcSMZ$ metabolized from SMZ was very fast in the plasma of the rabbit The elimination of $N_4AcSMZ$ was prolonged as compared to that of the parent drug Vie found no sex difference in the elimination pattern of $N_4AcSMZ$ in the rabbit.
Purpose: The preferred material for bone augmentation beyond the envelope of skeletal bone is the bone block graft, due to its dimensional stability. We evaluated the necessity of rigid fixation for the bone block graft, and compared the bone regeneration and volume maintenance associated with grafting using a synthetic hydroxyapatite block (HAB) and an autogenous bone block (ABB) without rigid fixation on rabbit calvaria over two different periods. Methods: Cylinder-shaped synthetic HAB and ABB were positioned without fixation on the rabbit calvarium (n=16). The animals were sacrificed at 4 or 8 weeks postoperatively, and the grafted materials were analyzed at each healing period using microcomputed tomography and histologic evaluation. Results: Integration of the graft and the recipient bed was observed in all specimens, although minor dislocation of the graft materials from the original position was evident in some specimens (six ABB and ten HAB samples). A tendency toward progressive bone resorption was observed in the grafted ABB but not in the grafted HAB, which maintained an intact appearance. In the HAB group, the area of new bone increased between 4 and 8 weeks postoperatively, but the difference was not statistically significant. Conclusions: The nonfixed HAB was successfully integrated into the recipient bed after both healing periods in the rabbit calvaria. In spite of limited bone formation activity in comparison to ABB, HAB may be a favorable substitute osteoconductive bone material.
Kang, T. Y.;Yin, X. J.;Rho, G. J.;Lee, H.;Chae, Y. J.;Lee, H. J
Journal of Embryo Transfer
/
v.15
no.2
/
pp.167-173
/
2000
The principal objective of this study was to clone transgenic embryos in order to improve the efficiency of transgenic animal production by the combination of microinjection and nuclear transplantation techniques. Mature female New Zealand White rabbits were superovulated by eCG and hCG treatments, fllowed by natural mating. Zygotes were collected from the oviducts at 18∼22 h after hCG injection by flushing with D-PBS containing 5% fetal calf serum(FCS). Two to three picoliters of green fluorescent protein(GFP) gene wa microinjected into male pronucleus. The foreign gene-injected zygotes were cultured in TCM-199 or RD medium containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% CO2 incubator. The morulae expressing GFP gene were selected and their blastomeres were separated for the use of nuclear donor. Following nuclear transplantation of fluorescence-positive morula stage blastomeres, 13 (21.3%) out of 61 fused oocytes developed to blastocyst stage and all of the cloned blastocysts expressed GFP. The results indicate that the screening of transgene in rabbit embryos by GFP detection could be a promisible method for the preselection of transgenic embryos. Also the cloning of preselected transgenic embryos by nuclear transplantatin could be efficiently applied to the multiple production of transgenic animals.
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