Seo, Hee-Jung;Lee, Seong-Kyu;Baik, Haing-Woon;Lee, Ki-Ho
Journal of Animal Science and Technology
/
v.54
no.3
/
pp.157-163
/
2012
The male reproduction is precisely controlled by a number of intrinsic and extrinsic factors. These factors usually involve in expressional regulation of various molecules influencing on sperm production in the testis. A number of ways are employed to control the transcription of specific genes, including epigenetic modifications of DNA and histone molecules. DNA methylation of CpG dinucleotides is a commonly used regulatory mechanism for testicular genes associated with the fertility. Previous studies have demonstrated the infertility induced by improper DNA methylation of these genes. In the present research, we attempted to determine transcriptional expression of some of these genes in the rat testis at different postnatal ages using real-time PCR analysis. These genes include neurotrophin 3 (Ntf3), insulin-like growth factor II (Igf2), JmjC-domain-containing histone demethylase 2A 1 (Jhm2da), paired box 8 transcription factor (Pax8), small nuclear ribonucleoprotein polypeptide N (Snrpn), and 5,10-methylenetetrahydrofolate reductase (Mthfr). The expression levels of Ntf3, Igf2, and Snrpn genes were the highest at the neonatal age, followed by transient decreases at the prepubertal age. Expression of Jhm2da and Mthfr genes were continuously increased from the neonate to 1 year of age. The levels of Pax8 mRNA at the early ages were higher than those at the later ages of postnatal development. These findings suggest that expression of some fertility-associated testicular genes in the rat during postnatal period could be differentially regulated by the control of the degree of DNA methylation.
Objective: This study investigated the effects of intrauterine growth restriction (IUGR) during late pregnancy on the cell growth, proliferation, and differentiation in ovine fetal thymuses. Methods: Eighteen time-mated Mongolian ewes with singleton fetuses were allocated to three groups at d 90 of pregnancy: restricted group 1 (RG1, 0.18 MJ ME/body weight [BW]0.75/d, n = 6), restricted group 2 (RG2, 0.33 MJ ME/BW0.75/d, n = 6) and control group (CG, ad libitum, 0.67 MJ ME/BW0.75/d, n = 6). Fetuses were recovered at slaughter on d 140. Results: The G0/G1 phase cell number in fetal thymus of the RG1 group was increased but the proliferation index and the expression of proliferating cell nuclear antigen (PCNA) were reduced compared with the CG group (p<0.05). Fetuses in the RG1 group exhibited decreased growth hormone receptor (GHR), insulin-like growth factor 2 receptor (IGF-2R), and their mRNA expressions (p<0.05). For the RG2 fetuses, there were no differences in the proliferation index and PCNA expression (p>0.05), but growth hormone (GH) and the mRNA expression of GHR were lower than those of the CG group (p<0.05). The thymic mRNA expressions of cyclin-dependent protein kinases (CDKs including CDK1, CDK2, and CDK4), CCNE, E2-factors (E2F1, E2F2, and E2F5) were reduced in the RG1 and RG2 groups (p<0.05), and decreased mRNA expressions of E2F4, CCNA, CCNB, and CCND were occurred in the RG1 fetuses (p<0.05). The decreased E-cadherin (E-cad) as a marker for epithelial-mesenchymal transition (EMT) was found in the RG1 and RG2 groups (p<0.05), but the OB-cadherin which is a marker for activated fibroblasts was increased in fetal thymus of the RG1 group (p<0.05). Conclusion: These results indicate that weakened GH/IGF signaling system repressed the cell cycle progression in G0/G1 phase in IUGR fetal thymus, but the switch from reduced E-cad to increased OB-cadherin suggests that transdifferentiation process of EMT associated with fibrogenesis was strengthened. The impaired cell growth, retarded proliferation and modified differentiation were responsible for impaired maturation of IUGR fetal thymus.
The present study was carried out to investigate the effects of co-culture cells and growth factors on in vitro culture of Korean native cattle(KNC) embryos fertilized in vitro. Two-eight cell embryos were cultured in vitro using 4 types of co-culture cells and 3 growth factors singly or in combination. The results were as follows, In the co-culture of 2~8 cell embryos with bovine oviductal epithelial cell(BOEC), granulosa cell(BGC), uterine epithelial cell(BUEC) and mouse embryonic fibroblast (MEF) monolayers, the developing rate to blastocysts was significantly(P<0.05) higher with BUEC(32.1%) than with MEF(15.3%), BGC(13.2%) and non co-culture control(11.6%). When the morula co-cultured with BOEC for 5 days following in vitro fertilization were co-cultured with BOEC continuously or with BUEC, respectively, the developing rate to blastocysts was higher with BUEC(73.9%) than with BOEC(56.0%). To examine the effects of growth factors on in vitro development of 2~8 cell embryos, epidermal growth factor(EGF), transforming growth factor-$\beta$l(TGF-$\beta$l) and insulin-like growth factor-1(IGF-1) were added singly or in combination to TCM 199 maturation medium with respective concentration. In a addition of each 10, 30 and SOng /rnl EGF, the developing rate to blastocysts was the highest in lOng /ml EGF(25.3%). In addition of each 1, 2 and Sng /mi TGF-$\beta$1, the developing rate to blastocysts was the highest in lng /ml TGF-$\beta$1(28.8%). In addition of each 50, 100ng/ml JGF-l, the developing rate to blastocysts was higher in 100ng/ml IGF-l(16.5%) than in SOng/mi IGF-1(12.9%). When lOng /ml EGF and lng /ml TGF-$\beta$l was added singly or in combination, the developing rate to blastocysts was similar in groups added singly or in combination with EGF and TGF-$\beta$l (23.l~24.6%), although higher than in control(16.7%). In the co-culture of 2~8 cell embryos Wth BOEC + each 10, 30 and 5Ong /rnl EGF, the developing rate to blastocysts was significantly(p<0.05) higher in BOEC + long /ml EGF(32.3%) than in BOEC + 3Ong /ml EGF(18.9%) and BOEC + song /ml EGF(9.7%). In the co-culture of 2~8 cell embryos with BOEC + each 1, 2, Sng /ml TGF-$\beta$l the developing rate to blastocysts was higher in BOEC + Sng/rnl TGF-$\beta$l(28.2%) than in BOEC + lng /ml TGF-$\beta$l(21.7%) and BOEC + 2ng/ml TGF-$\beta$l(21.4%). In summary, higher developing rate to blastocysts were obtained with co-culture of BUEC for co-culture system, with addition of lOng /ml EGF or lng /ml TGF-$\beta$l for growth factor culture system, and with co-culture of BOEC + lOng /ml EGF or BOEC + Sng /ml TGF-$\beta$l for co-culture + growth factor culture system.
The induction of nephric duct from Xenopus presumptive ectoderm(animal cap) was studied and the high-dose ef-fect of IGF-1 was investigated. Activin A induce various organs from cultured animal cap explants and the effects are time and dose-dependent. On the induction of nephric duct, the combined-dose of activin A and retinoic acid was very efficient method in reference study. In present study, we used IGF-1 as well as activin A as a combined growth factor. The concentration ranges of growth factors were activin A l00ng/ml an IGF-1 0-500ng/m1. Explants were cultured in combined solution for 3days to the normal embryo arrives at st. 43. In general, when the explant was cultured in high concentration(l00ng/ml) of activin A, it was destroyed, however, nephric duct and other tis-sues were differentiated by adding IGF-1. In addition, eye induction by adding IGF-1 500ng/ml to activin A 1- 100 ng/ml solution was studied. The low concentration of activin A(1ng/ml) have blood-like cell inducing effect and the explant was balloon-shaped, however, the high dose combination with IGF-1 extended the range of eye inductive concentration of activin A.
Kim, Min Ho;Kang, Moon Sung;Ha, Duck Min;Ko, Yong;Lee, C. Young
Journal of Animal Science and Technology
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v.55
no.2
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pp.95-101
/
2013
The present study was conducted to test a hypothesis that pork quality traits would be influenced by the systemic and/or local bioavailability of insulin-like growth factor-I (IGF-I), transforming growth factor-${\beta}1$ (TGF-${\beta}1$), or epidermal growth factor (EGF) before or at slaughter. To this end, 60 cross-bred female market pigs weighing approximately 110 kg were slaughtered, after which Longissimus dorsi muscle (LM) samples taken at slaughter (D 0) and blood samples taken at D -7 and D 0 were analyzed. The 60 carcasses rendered 36 RFN (reddish-pink, firm, and non-exudative), 16 RSE (reddish-pink, soft, and exudative), and 6 PSE (pale, soft, and exudative); 2 DFD (dark, firm, and dry) also were found but were excluded in subsequent experiments. The $L^*$ and drip loss were greater in PSE vs. RFN and RSE and in PSE and RSE vs. RFN, respectively, as they should (P<0.05). The $pH_{45min}$ was less in PSE vs. RFN (P<0.05); $pH_{24h}$ tended to be less in the former (P=0.09). The LM IGF-I and TGF-${\beta}1$ as well as serum EGF concentrations were less in PSE than in RFN. None of the other LM and serum concentrations of the three growth factors differed across the three pork quality categories. The LM IGF-I and TGF-${\beta}1$ concentrations and serum EGF concentration at D 0 were negatively correlated with drip loss [r = -0.36(P<0.01), -0.44 (P<0.01), and -0.32 (P<0.05), respectively]. However, none of the serum and LM growth factor variables was correlated with $L^*$ or $a^*$ (redness) of LM. Taken together, results suggest that locally expressed IGF-I and TGF-${\beta}1$ and blood-borne EGF may have a beneficial effect on postmortem water holding capacity of the muscle and that pork quality traits could be predicted to some extent from concentrations of IGF-I and TGF-${\beta}1$ in muscle and EGF in serum at slaughter.
Soo Hyun Kim;Kyu-Sang Sim;Jung Yoon Cheon;Jae-Woong Jang;Su Jin Jeong;Ye Hei Seo;Hye Myoung Ahn;Bong-Geun Song;Gi-Seok Kwon;Jung-Bok Lee
Journal of Life Science
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v.33
no.3
/
pp.234-241
/
2023
At present, many countries around the world are legalizing cannabis and its products, and research on various treatments using cannabis is being actively conducted. However, the cannabis plant contains other compounds whose biological effects have not yet been established. We investigated the effect of cannabidiol (CBD) on hair growth in human dermal papilla cells (HDPCs). 2,2'-Azino-bis (3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays were performed to determine the antioxidant activity of CBD. The HDPCs viability of CBD was examined via water-soluble tetrazolium salt (WST-1) assay. The expression of hair-loss-related markers in HDPCs by CBD treatment was analyzed by real-time PCR and western blotting. The DPPH, ABTS radical scavenging activity assay showed that CBD had superior antioxidant activities. In HDPCs, CBD increased cellular proliferation at concentrations without cytotoxicity. It also increased the expressions of fibroblast growth factor 1 (FGF1), fibroblast growth factor 7 (FGF7), vascular endothelial growth factor (VEGF), and insulin-like growth factor (IGF). These results correlated with a decrease in the expression of inhibition-related factors, such as androgen receptor (AR) and transforming growth factor beta 1 (TGF-B1). Moreover, CBD resulted in a significant increase in the phosphorylation of AKT and extracellular signal-regulated kinase (ERK). Therefore, it is suggested that CBD may be a potential remedy for the treatment of alopecia.
This experiment was designed to evaluate the effect of transforming growth factor-$\beta$ (TGF-$\beta$) and insulin-like growth factor-I (IGF-I) in bovine oocyte maturation in the presence or absence of serum on subsequent fertilization and embryo development. In addition, various concent rations of these growth factors were evaluated for the ability to promote development of eight-cell stage embryos to the blastocyst stage. Cumulus-oocyte complexes were recovered from 2 to 6 mm follicles obtained from slaughterhouse ovaries and cultured at 38.5$^{\circ}C$ for 24 hours in TCM-199 (HEPES Modification) with or with out 20 % fetal bovine serum (FBS) to which the following growth factors were added TGF$\beta$ IGF-l or TGF $\beta$ + IGF-I, all at 10 ng/ml each. The matured oocytes were fertilized in IVF-TL medium with frozen-thawed semen at a concentration of 1 ${\times}$ 10$^6$ cells/ml of fertilization medium following Percoll separation. After 24 hours of sperm-egg incubation, the embryos were transferred to CZB medium without glucose for 48 hours and then cultured in TCM-199 with 20% fetal bovine serum (FBS) for 96 hours. The addition of growth factors to IVM medium in the presence of serum had no effect on cleavage and subsequent embryo devlopment to blastocyst. In the absence of serum, TGF- improved cleavage and development to blastocyst compared to control's(p<0.05) and no synergistic effeet of IGF-I + TGF-$\beta$ was observed. In the second experiment, eight-cell embryos obtained by in vitro maturation (IVM) in TCM-199 + 20% FBS without growth facrors and in vitro fertil-ization (IVF) were cultured in the in vitro cuiture (IVC) medium supplemented with 5, 10 ng/ml TGF-$\beta$ or 5, 10, 50, 100 ng/ml IGF-I. Cleavage rate and development to the blastocyst stage was observed during seven days of incubation. The supplementation of 10 ng/ml TGF-$\beta$ to lVC medium for eight-cell embryos improved development to blastocyst (p<0.05) compared to control. In conclusion, these data indicate that the supplementation of growth factors to IVM medium in the presence of serum does not influence cleavage and subsequent embryo development. However, significantly more oocytes matured in serum-free TCM-199 and eight-cell embryos cultured in lVC medium developed to blastocyst with supplementation of 10ng/ml TGF-$\beta$.
Park, Joung-Hyun;Kim, Dong-Seok;Lee, Bae-Jin;Her, Jung-Soo;Jeon, Byeong Hwan
Journal of Marine Bioscience and Biotechnology
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v.13
no.2
/
pp.76-85
/
2021
Crassostrea gigas were fermented using L. brevis BJ20 to prepare fermented oyster extract (FO). The participants of this study were randomly assigned to FO and placebo (CON) groups. The FO group was given 1.0 g of FO supplementation and the CON group was given sucrose each day for eight weeks. The effects of FO supplementation on body composition, muscula r strength, and blood factors associated with muscle growth were assessed. The FO supplement was enriched with arginine (6,183.3 mg), phenylalanine (217.9 mg), leucine (122.6 mg), isoleucine (59.8 mg), valine (16.4 mg), and γ-amino butyric acid (GABA, 1,053.7 mg). The total fat was significantly decreased in the FO group compared with the CON group (p < 0.05). 60D/S Ext.T/Wo rk and 60D/S Flex.T/Work concomitantly with 60D/S Flex.PeakTQ/BW were significantly increase d by FO treatment compared to CON group (p < 0.05). However, posture stability was not significa ntly different between the groups. The levels of angiotensin-converting enzyme were significantly decreased within the FO group (p < 0.05). The FO group showed significantly decreased levels of tumor necrosis factor-α and increased levels of human growth hormone compared with the CON group (p < 0.01). The levels of insulin-like growth factor-1 increased (p < 0.01) in the FO group while that of creatine kinase and triglyceride decreased significantly compared with the CON group (p < 0.05). These results demonstrated that FO supplementation is effective in preventing sarcopenic obesity and maintaining and strengthening muscular function in elderly wom en. Hence, FO supplements can be used as functional ingredients for these benefits.
Engineered bone tissue is thought to be the ideal alternative for bone grafts in the treatment of related bone diseases. BMP9 has been demonstrated as one of the most osteogenic factors, and enhancement of BMP9-induced osteogenesis will greatly accelerate the development of bone tissue engineering. Here, we investigated the effect of insulin-like growth factor 1 (IGF1) on BMP9-induced osteogenic differentiation, and unveiled a possible molecular mechanism underling this process. We found that IGF1 and BMP9 are both detectable in mesenchymal stem cells (MSCs). Exogenous expression of IGF1 potentiates BMP9-induced alkaline phosphatase (ALP), matrix mineralization, and ectopic bone formation. Similarly, IGF1 enhances BMP9-induced endochondral ossification. Mechanistically, we found that IGF1 increases BMP9-induced activation of BMP/Smad signaling in MSCs. Our findings demonstrate that IGF1 can enhance BMP9-induced osteogenic differentiation in MSCs, and that this effect may be mediated by the enhancement of the BMP/Smad signaling transduction triggered by BMP9.
Objective: Endometrial cancer (EC) is the most common gynecologic malignancy. Identification of potential biomarkers of EC would be helpful for the detection and monitoring of malignancy, improving clinical outcomes. Methods: The Weighted Gene Co-expression Network Analysis method was used to identify prognostic markers for EC in this study. Moreover, underlying molecular mechanisms were characterized by KEGG pathway enrichment and transcriptional regulation analyses. Results: Seven gene co-expression modules were obtained, but only the turquoise module was positively related with EC stage. Among the genes in the turquoise module, COL5A2 (collagen, type V, alpha 2) could be regulated by PBX (pre-B-cell leukemia homeobox 1)1/2 and HOXB1(homeobox B1) transcription factors to be involved in the focal adhesion pathway; CENP-E (centromere protein E, 312kDa) by E2F4 (E2F transcription factor 4, p107/p130-binding); MYCN (v-myc myelocytomatosis viral related oncogene, neuroblastoma derived [avian]) by PAX5 (paired box 5); and BCL-2 (B-cell CLL/lymphoma 2) and IGFBP-6 (insulin-like growth factor binding protein 6) by GLI1. They were predicted to be associated with EC progression via Hedgehog signaling and other cancer related-pathways. Conclusions: These data on transcriptional regulation may provide a better understanding of molecular mechanisms and clues to potential therapeutic targets in the treatment of EC.
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