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Immunohistochemical study on the Bovine chromogranin, Serotonin and Bovine pancreatic polypeptide immunoreactive cells in the pancreas of the duck(Anas platyrhynchos platyrhyncos, Linne) (청둥오리 췌장에서 bovine chromogranin, serotonin 및 bovine pancreatic polypeptide 면역반응세포의 분포에 관한 연구)

  • Lee, Jae-hyun;Ku, Sae-kwang;Lee, Hyeung-sik
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.232-238
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    • 1998
  • The distributions and relative frequencies of bovine chromogranin(BCG)-, serotonin-, and bovine pancreatic polypeptide(BPP)-immunoreactive cells in the pancreas of the duck(Anas plaryrhynchos plaryrhyncos, Linne) were studied immunohistochemically on 23 days of incubation, at hatching, 1 week, 2 weeks, 3 weeks, 5 weeks, 6 weeks, 7 weeks, 9 weeks, 10 weeks, and 32 weeks after hatching. In the exocrine portions, the relative frequencies of BCG- and serotonin-immunoreactive cells tend to decrease according to ages. BPP-immunoreactive cells first appeared 1 week after hatching, and thereafter decreased. These cells were mainly observed in the exocrine pancreas and marginal region of the pancreatic islets. In the endocrine portions, BCG-, serotonin- and BPP-immunoreacteve cells disappeared 2 weeks, 9 weeks and 5 weeks after hatching, respectively.

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EFFECT OF TRYPSIN-DIGESTED BOVINE GROWTH HORMONE ON WHOLE-BODY PROTEIN SYNTHESIS IN VITRO IN CHICKEN EMBRYOS

  • Kita, K.;Hatano, S.;Okumura, J.;Muramatsu, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.2
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    • pp.319-323
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    • 1993
  • The effect of bovine growth hormone digested with trypsin on whole-body protein synthesis in vitro of chicken embryos was investigated by using a whole-embryo culture system. Bovine growth hormone at 5.3 and 530 ng/ml was digested partially and completely with trypsin for 4 min and 18 h, respectively. After culturing chicken embryos with a synthetic medium containing $L-[4-^3H]$ pheylalanine, whole-embryo protein synthesis was determined from the ratio of specific radioactivities of free and protein-bound pheylalanine. Whole-embryo protein synthesis of the control group cultured with no bovine growth hormone was $49.5{\pm}2.2%/d$. There was no significant interaction between digestion time and the concentration of trypsin-digested bovine growth hormone. Tryptic digestion of bovine growth hormone increased fractional synthesis rates of whole-body protein compared to the 0-min groups, and there was no significant difference between the 4-min and 18-h groups. The higher concentration (530 ng/ml) of trypsin-digested bovine growth hormone was more effective in enhancing whole-embryo protein synthesis than the lower concentration (5.3 ng/ml).

Production of Normal Calves after Transfer of IVF-Derived Bovine Embryos (체외수정란 유래의 송아지 생산)

  • 한용만
    • Korean Journal of Animal Reproduction
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    • v.18 no.1
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    • pp.7-13
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    • 1994
  • To verify in vivo viability of IVF-derived bovine embryos, morula and blastocysts that developed from in vitro matured and fertilized ova were transferred to the uteri of recipient cows and normal calves were produced. To produce IVF-derived bovine morula or blastocysts, ova matured and fertilized in vitro were cultured in culture medium for 7~8 days at 39$^{\circ}C$ under the humicified atmosphere of 5% CO2. Two different culture systems, a co-culture system with TCM-199 and bovine epithelial cells (BOEC) and CR1aa without somatic cell support, were compared. Cleavage rates to 2~8 cell stage and developmental rates of IVF-derived bovine embryos to blastocyst stage were not different between co-culture system (51.3 and 14.0%) and CR1aa medium (60.4 and 22.1%), respectively. Embryos were classified into three grades by embryo quality and then one or two embryos in higher quality(A and B grades) were transferred to the uterus of recipients. In this study Korean Native calf was first born after transfer of IVF-derived embryos. Total four live calves were normally developed to term from IVF-derived bovine blastocysts and one female fetus was still-born approximatedly 8 months of gestation, but there was no pregnancy after transfer of morula. Therefore, normal calves could be produced after transfer of IVF-derived bovine embryos cultured in CR1aa medium without somatic cell support. In addition, our results suggest that in transfer of IVF-derived bovine embryos blastocyst stage is better than morula.

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Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence In Situ Hybridization Technique

  • Han, M.S.;Cho, E.J.;Ha, H.B.;Park, H.S.;Sohn, S.H.
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.133-137
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    • 2004
  • Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.

Comparative scanning electron microscope analysis of the enamel of permanent human, bovine and porcine teeth

  • Olek, Aneta;Klimek, Leszek;Boltacz-Rzepkowska, Elzbieta
    • Journal of Veterinary Science
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    • v.21 no.6
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    • pp.83.1-83.14
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    • 2020
  • Background: Bovine and porcine teeth are often used in in vitro experiments as substitutes of human teeth. Objectives: The aim of the present study was to perform a comparative analysis of enamel morphology of permanent human, bovine and porcine teeth under the scanning electron microscope. Methods: As many as 10 human, 10 bovine, and 10 porcine teeth were studied. All the teeth were sectioned and the halves were randomly divided into 2 groups according to the examined tissue (vestibular enamel at the mid-height of the dental crown and in the cervical area). Human and bovine enamel was etched for 15 sec and porcine enamel for 30 sec. The scanning electron microscope analysis was performed. The length and width of enamel prisms were determined with the "Met-Ilo" 1.1 computer program. Results: All enamel samples revealed the same etching pattern-Silverstone's type 2. Bovine enamel showed a similar porosity and the amount of interprismatic enamel compared to human enamel while the amount and width of interprismatic enamel bands in porcine enamel were evidently greater. The shape of the porcine prisms was visually similar to human prisms, although dimensions were significantly different. However, bovine prisms differed in form and appeared to be distinctly elongated. Conclusions: Reported findings indicate that the results of experimental studies carried out on bovine and porcine enamel should not be compared with the results obtained on human enamel.

12-Oxoeicosatetraenoic acid, a candidate signal for placenta separation, activates matrix metalloproteinase and induces apoptosis in bovine trophoblast cells

  • Hachiro Kamada
    • Animal Bioscience
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    • v.36 no.3
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    • pp.429-440
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    • 2023
  • Objective: 12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-KETE), a metabolite of arachidonic acid, is a strong candidate signal for placenta separation following calf discharge at delivery. In the present study, the effects of 12-KETE on bovine trophoblast cells were investigated to determine its function in the placentome at delivery. Methods: Bovine trophoblast cells derived from blastocysts were used. They were cocultured with or without fibroblasts derived from bovine placentome and/or bovine uterine epithelial cells. 12-KETE was added to the culture medium. Results: Bovine trophoblast cells contained binucleate cells and strongly expressed caudal type homeobox 2 (CDX-2) genes. Addition of 12-KETE to the trophoblast cell colony without feeder cells or that on a fibroblast monolayer induced rapid exfoliation of the colony. After 12-KETE addition, trophoblast cells emitted strong fluorescence caused by the degradation of dye-quenched collagen, indicating that 12-KETE activated matrix metalloproteinase of the trophoblast cells. Exfoliated cell colonies were stained with YOPRO-1, but not propidium iodide (PI). Moreover, DNA fragmentation and Bcl-2 associated X protein (Bax) gene (apoptosis stimulator) upregulation were observed in exfoliated cells, indicating that 12- KETE induced trophoblast cell apoptosis. These results were consistent with previous in vivo observations; however, even a lower concentration of 12-KETE activated trophoblast protease. Meanwhile, fibroblasts derived from the bovine placentome converted arachidonic acid to 12-KETE. Conclusion: These observations indicate that 12-KETE may serve as a signal for placenta separation at delivery.

Characterization of Bovine NANOG5'-flanking Region during Differentiation of Mouse Embryonic Stem Cells

  • Jang, Hye-Jeong;Park, Hwan Hee;Tran, Thi Thuy Linh;Lee, Hak-Kyo;Song, Ki-Duk;Lee, Woon Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.12
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    • pp.1721-1728
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    • 2015
  • Embryonic stem cells (ESCs) have been used as a powerful tool for research including gene manipulated animal models and the study of developmental gene regulation. Among the critical regulatory factors that maintain the pluripotency and self-renewal of undifferentiated ESCs, NANOG plays a very important role. Nevertheless, because pluripotency maintaining factors and specific markers for livestock ESCs have not yet been probed, few studies of the NANOG gene from domestic animals including bovine have been reported. Therefore, we chose mouse ESCs in order to understand and compare NANOG expression between bovine, human, and mouse during ESCs differentiation. We cloned a 600 bp (-420/+181) bovine NANOG 5'-flanking region, and tagged it with humanized recombinant green fluorescent protein (hrGFP) as a tracing reporter. Very high GFP expression for bovine NANOG promoter was observed in the mouse ESC line. GFP expression was monitored upon ESC differentiation and was gradually reduced along with differentiation toward neurons and adipocyte cells. Activity of bovine NANOG (-420/+181) promoter was compared with already known mouse and human NANOG promoters in mouse ESC and they were likely to show a similar pattern of regulation. In conclusion, bovine NANOG 5-flanking region functions in mouse ES cells and has characteristics similar to those of mouse and human. These results suggest that bovine gene function studied in mouse ES cells should be evaluated and extrapolated for application to characterization of bovine ES cells.

Cloning of Bovine Macrophage Colony-stimulating Factor

  • Kim, Tae-Yung;Kim, Cheol-Ho;Lee, Sang-Gil;Kang, Chung-Boo
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.892-897
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    • 2005
  • Macrophage colony-stimulating factor (M-CSF) is a growth factor required for growth and differentiation of mononuclear phagocyte lineage. Total and 16 poly (A) mRNA of bovine M-CSF were isolated from healthy bovine peripheral mononuclear cells stimulated by phobol 12-myristste 13-acetate (TPA). The more compatible cultured mononuclear cells were 5${\times}$10/ml for RNA isolation. TPA-activated mononuclear cells increased the level of M-CSF-mRNA more than concanavalin A (Con A) and lipopolysaccharide (LPS). The optimal analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) for14 Macrophage colonystimulating factor (M-CSF) as a growth factor required for bovine M-CSF was denaturation at 94$^{\circ}C$ for 1 minute, annealing at 57$^{\circ}C$ for 1 minute, extension at 72$^{\circ}C$ for 1 minute for 30 cycles. The size of cDNA of bovine M-CSF by RT-PCR was 774 base pairs. A 774 base pairs cDNA encoding bovine M-CSF was synthesized by reverse transcriptase polymerase chain reaction (RT-PCR). Ligated cDNA was transformed to competent cells and then plasmid isolation and digestion was performed. Molecular cloning and sequencing were performed for cDNA of bovine M-CSF. The size of cloned cDNA of bovine M-CSF was 774base pairs. The homology of base sequence and amino acid sequence was 88% and 86% compared with known human M-CSF, respectively. From a high degree of sequence similarity, the obtained cDNA of bovine M-CSF is thought be a specific gene of bovine M-CSF.

Stable Inheritance of Bovine $\beta$-Casein/Bovine Growth Hormone Fusion Gene in Transgenic Mice (형질전환 생쥐에서 Bovine $\beta$-Casein/Bovine Growth Hormone 재조합 유전자의 유전적 안정성에 관한 연구)

  • 최영희;오건봉;강용국;방남수;서길웅;이경광;이철상
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.237-244
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    • 1998
  • 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.

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Linkage Disequilibrium Estimation of Chinese Beef Simmental Cattle Using High-density SNP Panels

  • Zhu, M.;Zhu, B.;Wang, Y.H.;Wu, Y.;Xu, L.;Guo, L.P.;Yuan, Z.R.;Zhang, L.P.;Gao, X.;Gao, H.J.;Xu, S.Z.;Li, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.6
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    • pp.772-779
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    • 2013
  • Linkage disequilibrium (LD) plays an important role in genomic selection and mapping quantitative trait loci (QTL). In this study, the pattern of LD and effective population size ($N_e$) were investigated in Chinese beef Simmental cattle. A total of 640 bulls were genotyped with IlluminaBovinSNP50BeadChip and IlluminaBovinHDBeadChip. We estimated LD for each autosomal chromosome at the distance between two random SNPs of <0 to 25 kb, 25 to 50 kb, 50 to 100 kb, 100 to 500 kb, 0.5 to 1 Mb, 1 to 5 Mb and 5 to 10 Mb. The mean values of $r^2$ were 0.30, 0.16 and 0.08, when the separation between SNPs ranged from 0 to 25 kb to 50 to 100 kb and then to 0.5 to 1 Mb, respectively. The LD estimates decreased as the distance increased in SNP pairs, and increased with the increase of minor allelic frequency (MAF) and with the decrease of sample sizes. Estimates of effective population size for Chinese beef Simmental cattle decreased in the past generations and $N_e$ was 73 at five generations ago.