• Animal Bioscience
• /
• 제34권6호
• /
• pp.957-966
• /
• 2021

Objective: Ovarian follicular development, which dependent on the proliferation and differentiation of granulosa cells (GCs), is a complex biological process in which miRNA plays an important role. Our previous study showed that miR-458b-5p is associated with ovarian follicular development in chicken. The detailed function and molecular mechanism of miR-458b-5p in GCs is unclear. Methods: The luciferase reporter assay was used to verify the targeting relationship between miR-458b-5p and catenin beta-1 (CTNNB1), which is an important transcriptional regulatory factor of the Wnt/β-catenin pathway. The cell counting kit-8 (CCK-8) assay, flow cytometry with propidium iodide (PI) and annexin V-fluorescein isothiocyanate (FITC) labeling were applied to explore the effect of miR-458b-5p on proliferation, cell cycle and apoptosis of chicken GCs. Quantitative real-time polymerase chain reaction and Western blot were used to detect the mRNA and protein expression levels. Results: We demonstrated that the expression of miR-458b-5p and CTNNB1 showed the opposite relationship in GCs and theca cells of hierarchical follicles. The luciferase reporter assay confirmed that CTNNB1 is the direct target of miR-458b-5p. Using CCK-8 assay and flow cytometry with PI and Annexin V-FITC labeling, we observed that transfection with the miR-458b-5p mimics significantly reduced proliferation and has no effects on apoptosis of chicken GCs. In addition, miR-458b-5p decreased the mRNA and protein expression of CD44 molecule and matrix metallopeptidase 7, which are the downstream effectors of CTNNB1 in Wnt/β-Catenin pathway and play functional roles in cell proliferation. Conclusion: Taken together, the data indicate that miR-458b-5p regulates ovarian GCs proliferation through Wnt/β-catenin signaling pathway by targeting CTNNB1, suggesting that miR-458b-5p and its target gene CTNNB1 may potentially play a role in chicken ovarian follicular development.

• 한국대기환경학회지
• /
• 제31권3호
• /
• pp.223-238
• /
• 2015

The $PM_{2.1}$ was collected by LVCI (low volume cascade impactor) during Group-A Period (September 1990 to December 2012) and the $PM_{2.5}$ was collected by HVAS (high volume air sampler) during Group-B Period (September 2009 to April 2012) at Kyung Hee University, Global Campus located on the boarder of Yongin and Suwon. The 8 water-soluble ions ($Na^+$, $NH_4{^+}$, $K^+$, $Mg^{2+}$, $Ca^{2+}$, $Cl^-$, $ NO_3{^-}$, and $SO_4{^{2-}}$) were analyzed by IC, and the 14 inorganic elements (Al, Mn, Si, Fe, Cu, Pb, Cr, Ni, V, Cd, Ba, Zn, Ti, Ag) were analyzed by XRF and ICP-AES after performing proper pre-treatments of each sample filter. The average total mass fractions of $SO_4{^{2-}}$, $NO_3{^-}$, and $NH_4{^+}$+ to $PM_{2.5}$ samples during Group-B Period were 0.39 in normal days, 0.44 in haze days, and 0.27 in Asian dust days, respectively; however, the average total mass fractions of Al, Fe, and Si to $PM_{2.5}$ mass were 0.043 in normal days, 0.021 in haze days, and 0.036 in Asian dust days, respectively. Especially the concentration of Pb was significantly decreased during Group-B Period rather than during Group-A Period, while Cr and Ni was increased during Group-B Period. In this study, we intensively compared the annual and seasonal patterns of major chemical species among normal days, haze days, and Asian dust days. Further we developed mass fraction profiles by collecting episode cases of haze days and Asian dust days, which were consisting of 22 chemical species. Those profiles are considered to be useful when applying various receptor models and establishing air quality management plans near future.

• Journal of Microbiology and Biotechnology
• /
• 제20권3호
• /
• pp.518-524
• /
• 2010

A mannanase gene (man26B) was obtained from a sea bacterium, Paenibacillus sp. BME-14, through the constructed genomic library and inverse PCR. The gene of man26B had an open reading frame of 1,428 bp that encoded a peptide of 475- amino acid residues with a calculated molecular mass of 53 kDa. Man26B possessed two domains, a carbohydrate binding module (CBM) belonging to family 6 and a family 26 catalytic domain (CD) of glycosyl hydrolases, which showed the highest homology to Cel44C of P. polymyxa (60% identity). The optimum pH and temperature for enzymatic activity of Man26B were 4.5 and $60^{\circ}C$, respectively. The activity of Man26B was not affected by $Mg^{2+}$ and $Co^{2+}$, but was inhibited by $Hg^{2+},\;Ca^{2+},\;Cu^{2+},\;Mn^{2+},\;K^+,\;Na^+$, and $\beta$-mercaptoethanol, and slightly enhanced by $Pb^{2+}$ and $Zn^{2+}$. EDTA did not affect the activity of Man26B, which indicates that it does not require divalent ions to function. Man26B showed a high specific activity for LBG and konjac glucomannan, with $K_m,\;V_{max}$, and $k_{cat}$ values of 3.80 mg/ml, 91.70 ${\mu}mol$/min/mg protein, and 77.08/s, respectively, being observed when LBG was the substrate. Furthermore, deletion of the CBM6 domain increased the enzyme stability while enabling it to retain 80% and 60% of its initial activity after treatment at $80^{\circ}C$ and $90^{\circ}C$ for 30 min, respectively. This finding will be useful in industrial applications of Man26B, because of the harsh circumstances associated with such processes.