• Asian-Australasian Journal of Animal Sciences
• /
• v.27 no.5
• /
• pp.743-748
• /
• 2014

The objective of this study was to determine the molecular characteristics of the horse vascular endothelial growth factor alpha gene ($VEGF{\alpha}$) by constructing a phylogenetic tree, and to investigate gene expression profiles in tissues and blood leukocytes after exercise for development of suitable biomarkers. Using published amino acid sequences of other vertebrate species (human, chimpanzee, mouse, rat, cow, pig, chicken and dog), we constructed a phylogenetic tree which showed that equine $VEGF{\alpha}$ belonged to the same clade of the pig $VEGF{\alpha}$. Analysis for synonymous (Ks) and non-synonymous substitution ratios (Ka) revealed that the horse $VEGF{\alpha}$ underwent positive selection. RNA was extracted from blood samples before and after exercise and different tissue samples of three horses. Expression analyses using reverse transcription-polymerase chain reaction (RT-PCR) and quantitative-polymerase chain reaction (qPCR) showed ubiquitous expression of $VEGF{\alpha}$ mRNA in skeletal muscle, kidney, thyroid, lung, appendix, colon, spinal cord, and heart tissues. Analysis of differential expression of $VEGF{\alpha}$ gene in blood leukocytes after exercise indicated a unimodal pattern. These results will be useful in developing biomarkers that can predict the recovery capacity of racing horses.

• Korean Journal of Poultry Science
• /
• v.26 no.4
• /
• pp.217-236
• /
• 1999

Among polyunsaturated fatty acids (PURAs) targeted for manipulation in animal tissues (poultry eggs and meat), omega-3 PUFAs(n-3 PUFAs) are discussed in this review. 3 or 5% dietary menhaden oil (MO) supplemented layer diets was reported to increase docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) contents in the egg. MO at 1.5% also increased the deposition of up to 180mg total omega-3 fatty acids / yolk. Utilization of 5% ground flax seed (FS) resulted in similar total omega-3 fatty acid (FA) deposition as 1.5% MO. However, the basic feed formulations used in the Canadian feed industry usually include 10 to 20% FS in the egg laying diets. Recently several studies reported that addition of tocopherols in layer diets increased the tocopherol content more in the egg than any other tissue. One of reports said that 3.5% dietary oil with added tocopherols resulted in increasing tocopherol deposition and FA composition of the egg and other tissues. In the poultry meat, redfish meal (RM;4, 8, 12, 15 and 30% of diet) or redfish oil (RO;2.1 or 4.2% of diet) added to the practical corn-wheat-soybean based diets resulted in an increase in omega-3 FA and docosapentaenoic acid (DPA) contents in broiler meat lipids. Linseed oil (LO;1.0, 2.5, and 5.0% of broiler diet) supplemented in broiler diets also resulted in omega-3 FA and the ratio of omega-6 being significantly higher in poultry meat lipid than MO. Concern about fish flavor resulted in research about fish oil (FO) supplementation in broiler diets. Without the use of antioxidants, no more than 1.5% FO should be fed to broilers due to unacceptable orders from the chicken carcasses. One recent research project found that over 50mg/kg of vitamin E was required for maintaining the stability of unsaturated lipids in the meat. In regards to 'fishy'or 'crabby'taint in the eggs and poultry meat, poultry products remained acceptable when dietary fish oils were stabilized with antioxidants.

• Korean Journal of Poultry Science
• /
• v.16 no.3
• /
• pp.169-174
• /
• 1989

Culled laying hens used in this study were Arbor acres, which reared to S 35 days old on commerical formula feed for chicken. Liver, gizzard, breast and thigh muscles separated from each carcass, and total lipid was extracted and fractionated to neutral, phospho and glycolipid and then fatty acid composition were analyzed. Liver had the highest level of. total lipid, and breast tissue had the least among tissues tested. The neutral, phospho and glycolipid contents of total lipid had more thigh, breast and gizzard than other tissues, respectively. The major fatty acid in total and neutral lipid were palmitic, stearic, oleic and linoleic acid. And the major fatty acid in phospholipid was palmitic, stearic, oleic, linoleic, arachidonic and docosahexaenoic acid, and palmitic, palmitoleic, stearic, oleic and linoleic acid in case of glycolipid. The fatty acid contents of neutral, phospho and glycolipid in total lipid had more oleic, docosahexaenoic and linoleic acid than other lipid, respectively. Contents of unsaturated fatty acid of total and neutral lipid were comparatively high in thigh, and phospho and glycolipid were high in breast and liver, respectively. Contents of Polyunsaturated fatty acids were comparatively high in phospholipids than other lipids.

• Korean Journal of Poultry Science
• /
• v.27 no.2
• /
• pp.155-161
• /
• 2000

Unfortunately, there is no technique which is stable and repetitive to produce transgenic chicken, although various ways of gene transfer including PGC-and embryonic cell-mediated gene transfer, DNA microinjection, virus inoculation and sperm cells have been employed. The aims of this study were 세 develop and establish such a stable, repetitive and efficient way of gene transfer giving a faithful gene expression during development after the reconstruction of embryo in an UV-irradiated egg. A dual reporter plasmid (pJJ9), a fusion gene containing lacZ and GFP driven by a CMV promoter was used to exploit either merits of both reporting markers. lacZ with strong signal or GFP with vital marking. Electroporated embryonic blastodermal cells (EBCs) in the presence of the pJJ9 DNA faithfully showed 377 bp PCR product and lacZ or GFP expressions in the identical cells in vitro of in vivo. Furthermore, analyses of expression pattern of the foreign DNA demonstrated that microinjected EBCs cells into the UV-irradiated recipient egg should participate in normal developmental process, for example, proliferation and differentiation into various tissues. Thirty percentages of the manipulated eggs showed lacZ expression in their tissues. These results together with the specific procedures used in this study should facilitate avian transgenesis.

• Journal of Nutrition and Health
• /
• v.10 no.2
• /
• pp.18-26
• /
• 1977

Among animals, the rabbit is known to be affected most sensitively by dietary changes and to be most susceptible to atherosclerosis. The exact reason is still unknown as to whether the primary cause is intrinsic (tissue itself) or extrinsic such as a blood factor which could be influenced by various dietary means. It is of utmost importance to check the nutritional quality of rabbit meat before it is accepted and adapted as a daily food item. To evaluate nutritional quality of rabbit meat, studies on various aspects of lipid components were carried out in comparison with other animal meats such as beef, pork and chicken also included in the study was the question whether the cooking and storage conditions influence the composition of fatty acids and cholesterol level. Some results and findings are listed below: 1. The content of linoleic acid (18 : 2), one of the essential fatty acids, was much higher in rabbit meat compared to the other meats. The Percentages of this Polyunsaturated fatty acids, was much higher in rabbit meat compared to the other meats. The percentages of this polyunsaturated fatty acid in terms of total fatty acids were $37.3{\pm}3.7$, 5.9, 14.5, and 21.9% for rabbit, beef, pork, and chicken respectively. The degree of unsaturation was high not only in meat but also in liver and adipose tissue of rabbit. The values of iodine number, the indication of degree of unsaturation, were known to be $102{\sim}107$, $32{\sim}47$, $46{\sim}67$, and $55{\sim}77$ for rabbit, beef, pork, and chicken respectively. Such a high proportion of this polyunsaturated fatty acid contained in rabbit meat could be harmful due to their Peroxidation effect. 2. A small amount of lower (short chain) fatty acids was isolated from rabbit tussues, which were not observed in other animal's tissues. The significance of this small amount of short chain fatty acids contained in rabbit meat remained an open question. 3. The concentration of total cholesterol in rabbit meat was similar to that of otherr but the content of esterified cholesterol was higher in rabbit meat. This was probably due to the perference of cholesterol to esterify with unsaturated fatty acids. By roasting the percentage of Polyunsaturated fatty acids was decreased while saturated palmitic acid was proportionally increased. 4 The composition of fatty acids were affected more by dry heat than moist heat. More research should be pursued to improve methods of preservation and storage to prevent possible peroxidation and rancidity problems of rabbit meat. In the meantime, the public should be informed to eat fresh rabbit meat and not to store it for a long period of time. This study was supported by the Ministry of Science and Technology in Korea.

• Development and Reproduction
• /
• v.5 no.1
• /
• pp.81-88
• /
• 2001

Gonadotropin-releasing hormone (GnRH) has been known to play a role in the regulation of hCG secretion by human placenta. Recently, a gene encoding the second f개m of GnRH (GnRH-II) was identified in human. Herein, we demonstrate that GnRH-II is expressed in human placenta and assess GnRH-II expression by nested RT-PCR and immunohistochemistry in human placenta during the first trimester. We found that two altematively spliced transcripts of GnW-II mRNA were expressed in human placental tissues of first trimester and the shorter variant had a 21-bp deletion in GnRH-associated peptide (GAP). Immunoreactive GnRH-II was localized in both cytotrophoblastic and syncytiotrophoblastic cytoplasm. The immunostaining intensity was stronger in cytotrophoblast. Villous stromal cells also showed GnRH-II immunoreactiyiry. The results of our study report that the second isoform of GnRH (GnRH-II) is expressed in the first trimester human placenta and we suggest that GnRH-II may also play a regulatory role in maintenance of early pregnancy and hCG secretion in human placenta.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.7
• /
• pp.1037-1047
• /
• 2017

Objective: Despite an increasing number of investigations into the pathophysiology of necrotic enteritis (NE) disease, etiology of NE-associated diseases, and gene expression profiling of NE-affected tissues, the microRNA (miRNA) profiles of NE-affected poultry have been poorly studied. The aim of this study was to induce NE disease in the genetically disparate Fayoumi chicken lines, and to perform non-coding RNA sequencing in the intestinal mucosal layer. Methods: NE disease was induced in the Fayoumi chicken lines (M5.1 and M15.2), and non-coding RNA sequencing was performed in the intestinal mucosal layer of both NE-affected and uninfected chickens to examine the differential expression of miRNAs. Next, quantitative real-time polymerase chain reaction (real-time qPCR) was performed to further examine four miRNAs that showed the highest fold differences. Finally, bioinformatics analyses were performed to examine the four miRNAs target genes involvement in the signaling pathways, and to examine their interaction. Results: According to non-coding RNA sequencing, total 50 upregulated miRNAs and 26 downregulated miRNAs were detected in the NE-induced M5.1 chickens. While 32 upregulated miRNAs and 11 downregulated miRNAs were detected in the NE-induced M15.2 chickens. Results of real-time qPCR analysis on the four miRNAs (gga-miR-9-5p, gga-miR-20b-5p, ggamiR-196-5p, and gga-let-7d) were mostly correlated with the results of RNAseq. Overall, ggamiR-20b-5p was significantly downregulated in the NE-induced M5.1 chickens and this was associated with the upregulation of its top-ranking target gene, mitogen-activated protein kinase, kinase 2. Further bioinformatics analyses revealed that 45 of the gene targets of gga-miR-20b-5p were involved in signal transduction and immune system-related pathways, and 35 of these targets were predicted to interact with each other. Conclusion: Our study is a novel report of miRNA expression in Fayoumi chickens, and could be very useful in understanding the role of differentially expressed miRNAs in a NE disease model.

• Animal Bioscience
• /
• v.35 no.7
• /
• pp.964-974
• /
• 2022

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry and economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for studies on HPAIV resistance. Therefore, in this study, we investigated gene expression related to the mitogen-activated protein kinase (MAPK) signaling pathway by comparing non-infected, HPAI-infected resistant, and susceptible Ri chicken lines. Methods: Resistant (Mx/A; BF2/B21) and susceptible Ri chickens (Mx/G; BF2/B13) were selected by genotyping the Mx and BF2 genes. Then, the tracheal tissues of non-infected and HPAIV H5N1 infected chickens were collected for RNA sequencing. Results: A gene set overlapping test between the analyzed differentially expressed genes (DEGs) and functionally categorized genes was performed, including biological processes of the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. A total of 1,794 DEGs were observed between control and H5N1-infected resistant Ri chickens, 432 DEGs between control and infected susceptible Ri chickens, and 1,202 DEGs between infected susceptible and infected resistant Ri chickens. The expression levels of MAPK signaling pathway-related genes (including MyD88, NF-κB, AP-1, c-fos, Jun, JunD, MAX, c-Myc), cytokines (IL-1β, IL-6, IL-8), type I interferons (IFN-α, IFN-β), and IFN-stimulated genes (Mx1, CCL19, OASL, and PRK) were higher in H5N1-infected than in non-infected resistant Ri chickens. MyD88, Jun, JunD, MAX, cytokines, chemokines, IFNs, and IFN-stimulated expressed genes were higher in resistant-infected than in susceptible-infected Ri chickens. Conclusion: Resistant Ri chickens showed higher antiviral activity compared to susceptible Ri chickens, and H5N1-infected resistant Ri chickens had immune responses and antiviral activity (cytokines, chemokines, interferons, and IFN-stimulated genes), which may have been induced through the MAPK signaling pathway in response to H5N1 infection.

• Korean Journal of Veterinary Research
• /
• v.41 no.3
• /
• pp.373-380
• /
• 2001

Marek's disease virus (MDV) can cause skin lesions including inflammatory to tumorous. The phenotypical changes of lymphocytes infiltrating in the skin lesions induced by MDV were not clear. Therefore, the skin biopsies taken at weekly intervals for 8 weeks from the same specific-pathogen free chickens inoculated with Md/5 MDV were examined to analysis the phenotypical changes of lymphocytes. Histologically skin lesions progressed from initial inflammatory to late tumorous. Sequentially CD4+ T lymphocytes increased gradually in number from initial skin lesions and were major composition cells in the tumor lesions. Regardless of inflammatory or tumor lesions, CD8+ T cells and ${\gamma}{\delta}$ T cells infiltrated particularly in the dermis and subcutaneous on which MDV was actively replicated in the feather follicle epithelium(FFE). In addition, IgG bearing B lymphocytes in considerable number infiltrated in the dermis and subcutaneous tissues. From these results, the development of MDV-induced skin lesions was inflammatory following tumorous. In addition, each CD8+, ${\gamma}{\delta}$ and CD4+ T cells and B cell might act to protect MDV replication in the FFE or tumor cells which turned on lytic cycle.

• BMB Reports
• /
• v.29 no.2
• /
• pp.127-132
• /
• 1996

Germinal centers (GCs) are formed in peripheral lymphoid tissues in response to protein antigens. In order to see if immunoglobulin isotype switching takes place in GC B-cells, we isolated GC B-cells (PNA positive cells) from mouse popliteal lymph nodes by a flow cytometer after the staining of lymph node cells with PNA-FITC and anti-B220-PE, and determined the expression of ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA by RT-PCR. ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA were amplified specifically in cDNAs from hybridoma expressing IgG1 or splenocytes stimulated LPS plus IL-4. Germinal center B-cells formed in popliteal lymph nodes of mice immunized with chicken ovalbumin were isolated 7 days after immunization. We sorted GC B-cells five times. Immunoglobulin ${\gamma}1$ germline transcripts were expressed in germinal center B-cells in three out of five sorts whereas two out of five sorts did not express ${\gamma}1$ germline transcripts in GC B-cells. The contents of GC B-cells ranged from 5 to 7% of total lymph node cells in most flow cytometric analyses but those of two sorted cells which did not express ${\gamma}1$ germline transcripts were out of normal range. These results imply that isotype switching of immunoglobulins may take place in GCs.