To improve the efficiency of nuclear transplantation in the rabbit, this study were evaluated the influence of celly cycle of donor nuclei on the in vitro developmental potential in the nuclear transplant embryos. The embryos of 16-cell stage were collected from the mated does at 48h post-hCG injection and they were synchronized to G1 phase of 32-cell stage. Synchronization of the cell cylce of blastomeres were induced, first, using an microtubules polymerization inhibitor, 0.5$\mu\textrm{g}$/ml colcemid for 10h to arrest blastomeres in metaphase, and secondly, using a DNA synthesis inhibitor, 0.1$\mu\textrm{g}$/ml aphidicolin for 1.5 to 2h to cleave to 32-cell stage and arrest them in G1 phase. The separated G1 phase blastomeres of 32-cell stage were injectied into enucleated recipient cytoplasms by micromanipulation. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused and activated by electrical stimulation. The nuclear transplant embryos were co-cultured for 120h. In vitro cultured embryos were monitored every 24h to assess for development rate. After in vitro cultue for 120h, the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 dye for counting the number of blastomeres under a fluorescence microscopy. The cleavage rate of blastomeres from 16-cell stage stage rabbit embryos treated with colcemid for 10h or aphidicolin for 6h following colcemid for 10h were not significantly different. The electrofusion rate was similar by high in S and G1 phase donor nuclei as 80.6 and 79.1%, respectively. However, the nuclear transplant embryos using G1 phase donor nuclei were developed to blastocyst at high rate(60.3%) than those using S phase donor nuclei(26.0%). Moreover, the mean blastocyst stage were increased significantly(P<0.05) with the G1 phase donor nuclei(176.6 cells and 1.50 cycles), as compared with the S phase donor nuclei(136.6 cells and 1.42 cycles). These results show that the blastomeres of G1 phase were more successful as donor nuclei in the nuclear transplant procedure, compared with S phase.
Glucose is universal and essential fuel of energy metabolism and in the synthesis pathways of all mammalian cells. Glucose is the one of the major precursors of lactose synthesis using glycolysis result in producing milk fat and protein. During the milk fat synthesis, lipoprotein lipase (LPL) and CD36 are required for glucose uptake. Various morecules such as acyl-CoA synthetase 1 (ACSL1) activity of acetyl-CoA synthetase 2 (ACSS2), ACACA, FASN AGPAT6, GPAM, LPIN1 are closely related with milk fat synthesis. Additionally, glucose plays a major role for synthesizing lactose. Activations of lactose synthesize enzymes such as membranebound enzyme, beta-1,4-galactosyl transferase (B4GALT), glucose-6-phosphate dehydrogenase (G6PD) are changed by concentration of glucose in blood resulting change of amount of lactose production. Glucose transporters are a wide group of membrane proteins that facilitate the transport of glucose over a plasma membrane. There are 2 types of glucose transporters which consisted facilitative glucose transporters (GLUT); and sodium-dependent transport, mediated by the Na+/glucose cotransporters (SGLT). Among them, GLUT1, GLUT8, GLUT12, SGLT1, SGLT2 are main glucose transporters which involved in mammary gland development and milk synthesis. However, more studies are required for revealing clear mechanism and function of other unknown genes and transporters. Therefore, understanding of the mechanisms of glucose usage and its regulation in mammary gland is very essential for enhancing the glucose utilization in the mammary gland and improving dairy productivity and efficiency.
Gavojdian, D.;Budai, C.;Cziszter, L.T.;Csizmar, N.;Javor, A.;Kusza, S.
Asian-Australasian Journal of Animal Sciences
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제28권4호
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pp.599-603
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2015
The objective of the current pilot study was to evaluate the reproductive performance and health indicators in Dorper, White Dorper, and Tsigai breed ewes managed semi-intensively under European temperate conditions. A total of 544 ewe-year units were observed, with ewes (ranging from 1.5 to 8 years of age) managed under identical rearing conditions for a period of two consecutive production cycles (2012 through 2013 and 2013 through 2014). In general, significant ($p{\leq}0.001$) genotype-related disparities were found in occurrence rates for all health parameters taken into study. Clinical mastitis incidence was significantly lower ($p{\leq}0.05$) in Dorper (9.4%) and White Dorper (10.8%) breeds compared to that of Tsigai ewes (17.4%). Significant differences ($p{\leq}0.05$) for lameness were found between Dorper and Tsigai breeds, with occurrence rates of 8.0% and 2.9%, respectively. Incidence for pneumonia and abortion was not influenced (p>0.05) by the ewes' genotype. Litter size was significantly lower ($p{\leq}0.05$) in White Dorper breed than for Dorper and Tsigai ewes, of 1.21, 1.40, and 1.45, respectively. Conception rates and lambs survival were not affected (p>0.05) by genotype. Results suggest that South African Dorper and White Dorper sheep breeds have adapted well to the specific rearing conditions.
한국펄프종이공학회 1999년도 Proceedings of Pre-symposium of the 10th ISWPC
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pp.209-213
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1999
In the society of the 21\ulcorner century under multiphase media conditions, the rapidly glowing electronic media will replace the conventional paper media in a variety of areas. However, if human being still has an affinity for paper media and an instinct for hardcopy from electronic-based text or image, the new market will be created for the paper industry. To what extent the consumer choses paper media for output will depend upon the availability of functions of paper media appealing to human senses; i.e., "sensory functions of paper". As a whole, on-demand type personal as well as business communications will increase in the next century and this trend will lead certainly to a rapidly expanding "contents hardcopy market". The technological progress of the paper industry in the 21\ulcorner century depends upon the market needs for higher products quality and higher efficiency of manufacturing process as well as an endeavour to overcome constraints from forest resource, energy, and environmental issues. Under the conditions with above constraints, the paper media will be polarized into two categories; (1)paper for higher image reproduction capability for original image or text and (2)paper for lower reproduction but with higher appeals for human senses. To cope with these trends, psycho-physical analysis and a sensory engineering approach for developing new paper media is vitally required. Also newly emerged roles of paper physics in the multimedia age is pointed out associated with sensory functions of paper that are not well-understood so far.
To investigate the fidelity of transgene transmission and expression, we produced transgenic mice carrying bovine $\beta$-casein/bovine grwoth hormone(bGH) fusion gene and examined transmission efficiency and expression level of the transgene in the founders and their progeny. The transgene was composed of 1.8 kb bovine $\beta$-casein promoter and 2.1 kb bGH gene. Ten transgenic mice were produced. Milk and mammary gland were collected from eight transgenic lines at 10-day lactation and a, pp.ied to Western and Northern blot analyses. The bGH expression was detected in four of them. The concentrations of bGH in milk were highly variable from 4$\mu\textrm{g}$/ml to 600$\mu\textrm{g}$/ml depending on the lines. The bGH mRNA level in mammary gland was closely correlated with the bGH concentration in milk in each transgenic line. These results indicated that bGH transgene expression was a, pp.opriately regulated in the mammary gland and secreted into milk in transgenic mice. By using two transgenic lines(#2, #7) secreting a considerable amoung of bGH into their milk, the inferitance and maintenance of transgenic phenotype were assessed in successive four generations. The mean transmission frequencies of transgene in lines #2 and #7 were 34% and 40%, respectively. The bGH concentration in milk were 80, 240, 120, 60$\mu\textrm{g}$/ml in each G0(generation 0), G2, G3, G4 generation of line #2 and 600, 1600, 860, 900$\mu\textrm{g}$/ml in each G1. G2, G3, G4 generation of line #7. These results demonstrated that bovine $\beta$-casein/bGH gene was stably transmitted from generation to generation in a Menelian fashion in trasgenic mice and consistenly expressed in their milk throughout the generations, although there was a little variation in the transmission frequency and expression level of the transgene between generations.
To examine the efficiency of nuclear transplantation the influence of electrical preactivation of recipient cytoplasm on the in vitro developmental potentyl in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgery procedure. The separated G1 phase blastomeres of 32-cell stage were put into the non-preactivated and/or the preactivated recipient cytoplasm by electrical stimulation. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused. The fused nuclear transplant embryos were co-cultured with rabbit oviduct epithelial cells and monitored every 24h to assess for developmental rate. After in vitro culture for 120h, the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 and their blastomere were counted. The electrofusion rate was similar to the non-preactivated and preactivated recipient cytoplasm(81.8 and 85.7%, respectively). However, the in vitro developmental rate to blastocyst stage with the non-preactivated recipient cytoplasm (57.1%) was found significantly (P<0.05) higher, compared to the preactivated recipient cytoplasm(20.8%). The cell counts of nuclear transplant embryos developed to blastosyst stage were increased significantly (P<0.05) more in the non-preactivated recipient cytoplasm (163.7 cells), as compared with the preactivated recipient cytoplasm(85.4 cells). These results considered better that non-preactivated oocytes, MII phase oocytes, were used for recipient cytoplasms in the rabbit nuclear transplant procedure.
This experiment was undertaken to examine the effects of HIS treatment on the motility and acrosome reaction of frozen bovine spermatozoa and to test their abilities to interact with zona-free hamster eggs in vitro. Also, in vitro results were compared with those of bull's fertility in AI. The frozen semen from four Holstein bulls were exposed to HIS-DM for 5 minutes after thawing and then preincubated for 60 minutes in DM prior to insemination. The hamster eggs were mounted, fixed and stained 6 hours after exposure to boving spermatozoa and examined under a phase-contrast microscope. 1. The sperm motility expressed as a mobility index dro, pp.d significantly from 60-75 to 12-24 after exposure to HIS-DM, but increased in 32 to 41 at insemination. Bull C showed a low motility index than those of the orher bulls. The percentage of acrosome reaction by staining procedure were increased by HIS-DM treatment but did not change during 7 hours incubation period in DM. 2. The overall percentage of hamster eggs interacting with bull spermatozoa was 56.3%, 58.3%, 66.6% and 70.0%, respectively. Although there was no significant difference among bulls in the penetration rate of spermatozoa into hamster eggs, high proportions of eggs interacted with spermatozoa from Bull C and D than those from Bull A and B. 3. The conception rates (60-90 day RP) resulting from AI were 62.5%, 67.5% and 70.9% for Bull A, B and C, respectively. These results were in good agreement with the invitro results that the proportions of bull sperm-egg interction were greater for Bull C than for Bull A and B.
Kim, Doo-San;Hwangbo, Yong;Cheong, Hee-Tae;Park, Choon-Keun
한국동물생명공학회지
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제35권1호
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pp.58-64
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2020
This present study was conducted to investigate protective effect of discontinuous Percoll gradient containing alpha-linolenic acid (ALA) before freezing process on viability, acrosome damage, mitochondrial activity, and oxidative stress of frozen-thawed boar spermatozoa. The separation of spermatozoa by discontinuous Percoll gradient was performed by different concentration of Percoll solution (45/90%) containing ALA combined with bovine serum albumin (BSA), and collected sperm in each Percoll layer was cryopreserved. To evaluate viability, acrosome damage, mitochondrial activity, and reactive oxygen species (ROS) level of frozen-thawed sperm, flow cytometry was used. Morphological abnormalities were observed under light microscope. In results, viability of sperm from 90% Percoll layer was higher than control and 45% Percoll group (p < 0.05). Separated sperm in 90% Percoll layer had lower acrosome damage and morphological abnormalities than control as well as viability, whereas 45% Percoll group was higher (p < 0.05). Similar with acrosome damage and abnormalities, mitochondrial activity was slightly enhanced and the population of live sperm with high ROS level was decreased by 90% Percoll separation, however, there was no significant difference. Supplementation of 3 ng/mL ALA into Percoll solution increased sperm viability and decreased population of live sperm with high ROS compared to control (p < 0.05). In conclusion, discontinuous Percoll gradient before freezing process could improve efficiency of cryopreservation of boar sperm through selection of sperm with high freezing resistance, and supplement of ALA during Percoll gradient might contribute suppression of ROS generation via stabilizing of plasma membrane during cryopreservation.
These experiments were investigate the effects of bacterial infection of uterus and vagina during bovine embryo transferring on the development of embryo. We examined the distribution of reproductive disordered cow by akind of disease, identified the bacteria isolated from the vagina of those cows and bacterial infectin of media and its treatment with several kinds of antibiotics at that. The results obtained were summarized as follows ; 1. The total 592 reproductive disordered cows were caused by ovarian dysfunction(43.4%), ovary-uterus complication(24.5%), endometrities(17.7%), and repeat breeder(12.0%). 2. The main bacteria among 11 kinds of bacteria(113 colonies) was E. coli(38 colonies, 33.6%). Likewise, E. coli was propotioned to 23 and 22.7% among bacteria from vagina of endometritis and repeat breeder, respectively. 3. The sensitivities of viginal bacteria to pencillin and streptomycin were 6.2 and 4.4% respectively, but those to gentamycin and chloramphenicol were 22.1 and 16.8%, respectively. The similar sensitivities were found in the embryo recovery media. 4. The rates of bacterial infection of recovery medim and that of abnormal development of embryo were 75 and 80%, respectively. 5. The antibiotic sensitivity assay of ova recovery media showed gentamicin and chloramphenicol gave better results than streptomycin and penicillin. 6. The developmental rate of 1-cell stage mice embryos was 34.0% in bacterial infected culture media, but was 40.0, 58.0, 40.0 or 30.0% with the treatment of kanamycin, gentamycin, chloramphenicol, streptomycin, or penicillin, respectively.
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