• Title/Summary/Keyword: Mammary Development

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Characterizing Milk Production Related Genes in Holstein Using RNA-seq

  • Seo, Minseok;Lee, Hyun-Jeong;Kim, Kwondo;Caetano-Anolles, Kelsey;Jeong, Jin Young;Park, Sungkwon;Oh, Young Kyun;Cho, Seoae;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.343-351
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    • 2016
  • Although the chemical, physical, and nutritional properties of bovine milk have been extensively studied, only a few studies have attempted to characterize milk-synthesizing genes using RNA-seq data. RNA-seq data was collected from 21 Holstein samples, along with group information about milk production ability; milk yield; and protein, fat, and solid contents. Meta-analysis was employed in order to generally characterize genes related to milk production. In addition, we attempted to investigate the relationship between milk related traits, parity, and lactation period. We observed that milk fat is highly correlated with lactation period; this result indicates that this effect should be considered in the model in order to accurately detect milk production related genes. By employing our developed model, 271 genes were significantly (false discovery rate [FDR] adjusted p-value<0.1) detected as milk production related differentially expressed genes. Of these genes, five (albumin, nitric oxide synthase 3, RNA-binding region (RNP1, RRM) containing 3, secreted and transmembrane 1, and serine palmitoyltransferase, small subunit B) were technically validated using quantitative real-time polymerase chain reaction (qRT-PCR) in order to check the accuracy of RNA-seq analysis. Finally, 83 gene ontology biological processes including several blood vessel and mammary gland development related terms, were significantly detected using DAVID gene-set enrichment analysis. From these results, we observed that detected milk production related genes are highly enriched in the circulation system process and mammary gland related biological functions. In addition, we observed that detected genes including caveolin 1, mammary serum amyloid A3.2, lingual antimicrobial peptide, cathelicidin 4 (CATHL4), cathelicidin 6 (CATHL6) have been reported in other species as milk production related gene. For this reason, we concluded that our detected 271 genes would be strong candidates for determining milk production.

Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

Role of cAMP, EGF, IGF-I and Protein Phosphorylation in Mammary Development II. Interaction Effects of EGF, IGF-I and Photoreactive Cyclic AMP on DNA Synthesis and Protein Phosphorylation (유선발달에 있어서 cAMP, EGF, IGF-I 및 단백질 인산화 작용의 역할 II. EGF, IGF-I 및 Photoreactive Cyclic AMP의 상호작용과 단백질 인산화 작용)

  • 여인서
    • Korean Journal of Animal Reproduction
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    • v.19 no.2
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    • pp.95-104
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    • 1995
  • Mouse mammary epithelial cells(NMuMG) were maintained onto 6-well plates (3$\times$105 cells/well) or chambered slide (1$\times$104 cells/well), in DMEM supplemented with 10% fetal calf serum. After serum starvation for 24 hours, DMNB (1$\mu$M) was added and exposed to UV light (300nm, 3 second pulse) after 2 hours from DMNB addition in order to activate DMNB which induces a rapid transient increase in intracellular cAMP upon UV irradiation. EGF (100ng/ml) and/or IGF-I (10ng/ml) were treated at the time of UV irradiation. Nuclear labeling index was estimated as percent of nuclear labeled cells(percent of S phase of cells) by incorporation of 3H-thymidine into DNA(1 hour pulse with 1$\mu$Ci/ml). DMNB(1$\mu$M), EGF (100ng/ml) and/or IGF-I (10ng/ml) signifciantly increased nuclear labeling index than those of control (P<0.05). Addition of DMNB+EGF or DMNB+EGF+IGF-I showed the interaction effect in nuclear labeling index (P<0.05). Protein kinase A activities by addition of EGF, IGF-I or EGF+IGF-I were 10.5, 9.8 or 9.4 unit/mg protein, respectively, and no statistical difference was found in comparison with control (P>0.05). Additon of DMNB+EGF showed the moderate interaction effect on tyrosyl kinase activity (P<0.1). In the fluorography analysis, there were no specific protein phosphorylation patterns were found at 1 or 15 minute by addition of DMNB. EGF and/or IGF-I. These results suggest that the interaction effect in nuclear labeling index by addition DMNB and EGF could be mediated through the modulation of tyrosyl kinase activity by cAMP.

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Characterization of Double Transgenic Mice Harboring Both Goat $\beta$-casein/hGH and Goat $\beta$-casein/hG-CSF Hybrid Genes

  • Oh, Keon-Bong;Lee, Chul-Sang
    • Development and Reproduction
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    • v.13 no.3
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    • pp.191-198
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    • 2009
  • In an attempt to simultaneously produce two human proteins, hGH and hG-CSF, in the milk of transgenic mice, we constructed goat $\beta$-casein-directed hGH and hG-CSF expression cassettes individually and generated transgenic mice by co-injecting them into mouse zygotes. Out of 33 transgenic mice, 29 were identified as double transgenic harboring both transgenes on their genome. All analyzed double transgenic females secreted both hGH and hG-CSF in their milks. Concentrations ranged from 2.1 to $12.4\;mg/m{\ell}$ for hGH and from 0.04 to $0.13\;mg/m{\ell}$ for hG-CSF. hG-CSF level was much lower than hGH level but very similar to that of single hG-CSF mice, which were introduced with hG-CSF cassette alone. In order to address the causes of concentration difference between hGH and hG-CSF in milk, we examined mRNA level of hGH and hG-CSF in the mammary glands of double transgenic mice and tissue specificity of hG-CSF mRNA expression in both double and single transgenic mice. Likewise protein levels in milk, hGH mRNA level was much higher than hG-CSF mRNA, and hG-CSF mRNA expression was definitely specific to the mammary glands of both double and single transgenic mice. These results demonstrated that two transgenes have distinct transcriptional potentials without interaction each other in double transgenic mice although two transgenes co-integrated into same genomic sites and their expressions were directed by the same goat $\beta$-casein promoter. Therefore goat $\beta$-casein promoter is very useful for the multiple production of human proteins in the milk of transgenic animals.

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Development of Automatic Measurement of Body Temperature by Taking the Temperature of Milk while Milking in Dairy Cattle (젖소에서 유즙체온 측정을 이용한 체온 자동 측정 방법의 개발)

  • 김용준;한종현;이수영;한병성;김동원
    • Journal of Veterinary Clinics
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    • v.19 no.1
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    • pp.37-42
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    • 2002
  • To develop an automatic detecting system of body temperature of dairy cattle while milking, measurement of the temperature of mammary skin using three thermometers attached into the lining of teat cup was carried out for 23 dairy cattle, whereas measurement of the temperature of milk while milking was also performed for 263 animals. For the latter experiment, three thermometers were attached at 10cm(left and right) and 20 cm away from an individual milk collector on the milk transporting hose. Taking the rectal temperature was accompanied all the time for the experiments. The measurement of the temperature of mammary skin using teat cup was successful for 11 of 23 dairy cattle(47.8%) and the mean temperature was $33.5^{\circ}C$ with the mean difference of $5.2^{\circ}C$ from the mean rectal temperature. The measurement of the temperature of milk using the thermometers onto the milk transporting hose while milking was very successful , From 37.3 to $38.4^{\circ}C$ of rectal temperature, the temperature of milk was almost the same and from 38.5 to $39.5^{\circ}C$ of rectal temperature, the temperature of milk tended to be low with the difference of 0.1$^{\circ}C$. From 39.6 to $41^{\circ}C$ of rectal temperature, the temperature of milk tended to be low with the difference of $0.2-0.6^{\circ}C$. These results indicated that automatic detection of body temperature whether low or high can be possible if the temperature of milk is taken while milking and if it is connected to the integration system by on-line.

Coping with large litters: management effects on welfare and nursing capacity of the sow

  • Peltoniemi, Olli;Han, Taehee;Yun, Jinhyeon
    • Journal of Animal Science and Technology
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    • v.63 no.2
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    • pp.199-210
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    • 2021
  • A number of management issues can be used as drivers for change in order to improve animal welfare and nursing capacity of the hyperprolific sow. Group housing of sows during gestation is a recommended practice from the perspective of animal welfare. Related health issues include reproductive health and the locomotor system. It appears that management of pregnant sows in groups is challenging for a producer and considerable skill is required. We explored the benefits and challenges of group housing, including feeding issues. Increasing litter size requires additional attention to the mammary gland and its ability to provide sufficient nursing for the growing litter. We discuss the fundamentals of mammary development and the specific challenges related to the hyperprolific sow. We also address challenges with the farrowing environment. It appears that the old-fashioned farrowing crate is not only outdated in terms of welfare from the public's perspective, but also fails to provide the environment that the sow needs to support her physiology of farrowing, nursing, and maternal behaviour. Studies from our group and others indicate that providing the sow with a loose housing system adequate in space and nesting material, along with reasonable chance for isolation, can be considered as fundamental for successful farrowing of the hyperprolific sow. It has also been shown that management strategies, such as split suckling and cross fostering, are necessary to ensure proper colostrum intake for all piglets born alive in a large litter. We thus conclude that welfare and nursing capacity of the sow can be improved by management. However, current megatrends such as the climate change may change sow management and force the industry to rethink goals of breeding and, for instance, breeding for better resilience may need to be included as goals for the future.

Complex Regulatory Network of MicroRNAs, Transcription Factors, Gene Alterations in Adrenocortical Cancer

  • Zhang, Bo;Xu, Zhi-Wen;Wang, Kun-Hao;Lu, Tian-Cheng;Du, Ye
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2265-2268
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    • 2013
  • Several lines of evidence indicate that cancer is a multistep process. To survey the mechanisms involving gene alteration and miRNAs in adrenocortical cancer, we focused on transcriptional factors as a point of penetration to build a regulatory network. We derived three level networks: differentially expressed; related; and global. A topology network ws then set up for development of adrenocortical cancer. In this network, we found that some pathways with differentially expressed elements (genetic and miRNA) showed some self-adaption relations, such as EGFR. The differentially expressed elements partially uncovered mechanistic changes for adrenocortical cancer which should guide medical researchers to further achieve pertinent research.

Relationship between the Polymorphisms of 5' Regulation Region of Prolactin Gene and Milk Traits in Chinese Holstein Dairy Cows

  • Li, J.T.;Wang, A.H.;Chen, P.;Li, H.B.;Zhang, C.S.;Du, L.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.4
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    • pp.459-462
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    • 2006
  • Prolactin (PRL) plays an important role in promoting mammalian mammary gland development, and milk production during lactation. Therefore the PRL gene was chosen as a candidate gene for milk traits in Holstein dairy cows. PCR-SSCP and PCR-RFLP were used to analyze genetic variations in the 5' regulation region of the PRL gene. In this part of the gene, two new polymorphic sites were detected in the Chinese Holstein dairy cows. One was a XbaI-RFLP locus, and the other was an SSCP locus. Statistical analysis showed that the XbaI-RFLP locus and the SSCP locus had a significant positive effect on milk traits.

Receptor activator of nuclear factor-κB ligand in T cells and dendritic cells communication

  • Nam, Sun-Young;Jeong, Hyun-Ja
    • CELLMED
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    • v.3 no.1
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    • pp.3.1-3.3
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    • 2013
  • The receptor activator of NF-${\kappa}B$ ligand (RANKL), a member of the tumor necrosis factor ligand family, has extensive functions beyond osteoclast development. RANKL is expressed in many immune cells such as osteoblasts, osteocytes, marrow stromal cells, activated T cells, synovial cells, keratinocytes, and mammary gland epithelial cells as well as in various tissues. The ligation of RANK by RANKL promotes dendritic cells (DCs) survival through prosurvival signals and the up-regulation of the anti-apoptotic proteins Bcl-2 and Bcl-$x_L$ and plays a crucial role in DCs-mediated Th1 differentiation. Therefore, RANKL plays an important role in the regulation of DCs/T cells-mediated specific immunity. This review will briefly inform our current understanding of the role of RANKL signaling in T cells-DCs communication in the immune system.