Changes of alkaline phosphatase activity in response to different stressors in planarian Dugesia japonica

Biologia ◽  
2013 ◽  
Vol 68 (2) ◽  
Author(s):  
Guang-Wen Chen ◽  
Ke-Xue Ma ◽  
De-Zeng Liu
Keyword(s):  
Alkaline Phosphatase ◽  
Heat Shock ◽  
Early Stage ◽  
High Concentration ◽  
Experimental Conditions ◽  
Alp Activity ◽  
Planarian Regeneration ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Dugesia Japonica

AbstractThe aims of this work are to provide some properties of alkaline phosphatase (ALP) in the planarian Dugesia japonica and detect its activity in response to different stressors, as well as to introduce renatured SDS-PAGE to study enzyme activity in planarians. Our results indicate that ALPs in planarians are mainly membrane-bound form, identified as three mainly enzyme-bands (approximately MW 260 kD, 180 kD, 160 kD, respectively). Under our experimental conditions, ALPs activity had no apparent changes in response to low concentration of Hg2+ (25 μg L−1) and Pb2+ (125 μg L−1, 250 μg L−1) exposure, but were severely inhibited in response to high concentration of Hg2+ (50 μg L−1, 150 μg L−1, 300 μg L−1) and Pb2+ (500 μg L−1, 1000 μg L−1) exposure. Mild heat shock (25°C for 2 days) elevated ALP activity, but severely heat shock (25°C for 2 days, followed by 30°C for 2 days and 32°C for 2 days) inactivated ALP activity. Interestingly, ALP and other cytosolic phosphatases (MW from ∼45 kD to ∼60 kD) activity increased noticeably during the early stage of planarians regeneration, which may be involved in cell proliferation and differentiation. Contrary to regeneration, prolonged starvation suppressed ALP activity. The above findings provide valuable information about the role of ALP in planarian regeneration and for its use as an indicator in ecotoxicology.

Isolation and identification of peptides from Pinctada martensii with osteogenic activity

2021 ◽  
Keyword(s):  
Mass Spectrometry ◽  
Alkaline Phosphatase ◽  
Liquid Chromatography Mass Spectrometry ◽  
Osteoblast Proliferation ◽  
Isolation And Identification ◽  
Osteogenic Activity ◽  
Alp Activity ◽  
Pinctada Martensii ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation

Abstract Marine organisms have attracted considerable attention in recent years. In this study, peptides with osteogenic activity from Pinctada martensii were isolated and identified. Additionally, the effects of the hydrolysates on MC3T3-E1 cell proliferation and differentiation were evaluated using the MTT and alkaline phosphatase (ALP) assays, respectively. First, trypsin, pancreatin, and neutral protease were used to hydrolyse the intact shellfish. The hydrolysates with the greatest effects on osteoblast proliferation and ALP activity were separated and purified. Second, fraction WP2 was isolated and purified using a Sephadex G-25 column. WP2, which had the highest osteogenic activity, increased cell growth by 48.57 ± 0.05% and ALP activity by 6.27 ± 0.07 mU. Finally, four novel peptides were identified in WP2 (FDNEGKGKLPEEY, IVLDSGDGVTH, IVLDSGDGVSH, and SSENSDLQRQ) by Orbitrap Fusion Lumos Tribrid orbital liquid chromatography-mass spectrometry. Our findings revealed that P. martensii contains peptides with potential osteogenic activity.

scholarly journals Fish Collagen Promotes the Expression of Genes Related to Osteoblastic Activity

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mark Luigi Fabian Capati ◽  
Ayako Nakazono ◽  
Kohei Yamamoto ◽  
Kouji Sugimoto ◽  
Kajiro Yanagiguchi ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Clinical Application ◽  
Biological Effects ◽  
Pcr Analysis ◽  
Type I ◽  
Experimental Conditions ◽  
Suitable Alternative ◽  
Alkaline Phosphatase Staining ◽  
Proliferation And Differentiation

Tilapia type I atelocollagen (TAC) is a strong candidate for clinical application as its biological scaffold due to a high degeneration temperature and biologically safe properties. The aim of this study was to confirm the biological effects of TACin vitroon osteoblastic cells, simulating its clinical application. The proliferation and differentiation of typical preosteoblasts, MC3T3-E1 cells, were investigated using a microarray analysis, staining assay for mineralization, and real-time PCR analysis of the expression of mineralization-related genes. The mRNA expression of 10 genes involved in proliferation and differentiation increased after 3-day culture on an TAC gel, with an average balanced score ratio exceeding 1.5 compared to the control. After two weeks of culture, all three experimental groups showed stronger alkaline phosphatase staining than after one week. The genes expression of alkaline phosphatase, osteocalcin, and bone sialoprotein increased under the experimental conditions. The gene expression of osteopontin did not increase, and no statistical differences were noted among the three experimental groups. The present and previous findings suggest that TAC is not only a suitable alternative to collagen products originating from mammals but also a novel biomaterial with cell differentiation ability for regenerative medicine.

Effects of heat shock on neuroblastoma (N1E 115) cell proliferation and differentiation

1992 ◽  
Vol 200 (1) ◽  
pp. 89-96 ◽  
Author(s):  
Angela Stoklosinski ◽  
Hanno Kruse ◽  
Christiane Richter-Landsberg ◽  
Ludger Rensing
Keyword(s):  
Cell Proliferation ◽  
Heat Shock ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation

scholarly journals Clinical and Molecular Insights in Erythropoiesis Regulation of Signal Transduction Pathways in Myelodysplastic Syndromes and β-Thalassemia

2021 ◽  
Vol 22 (2) ◽  
pp. 827
Author(s):  
Sarah Parisi ◽  
Carlo Finelli ◽  
Antonietta Fazio ◽  
Alessia De Stefano ◽  
Sara Mongiorgi ◽  
...  
Keyword(s):  
Myelodysplastic Syndromes ◽  
Transforming Growth Factor ◽  
Early Stage ◽  
Hypoxia Inducible Factor ◽  
Phosphatidylinositol 3 Kinase ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
And Function ◽  
Preclinical And Clinical Studies ◽  
Role And Function

Erythropoiesis regulation is essential in normal physiology and pathology, particularly in myelodysplastic syndromes (MDS) and β-thalassemia. Several signaling transduction processes, including those regulated by inositides, are implicated in erythropoiesis, and the latest MDS or β-thalassemia preclinical and clinical studies are now based on their regulation. Among others, the main pathways involved are those regulated by transforming growth factor (TGF)-β, which negatively regulates erythrocyte differentiation and maturation, and erythropoietin (EPO), which acts on the early-stage erythropoiesis. Also small mother against decapentaplegic (SMAD) signaling molecules play a role in pathology, and activin receptor ligand traps are being investigated for future clinical applications. Even inositide-dependent signaling, which is important in the regulation of cell proliferation and differentiation, is specifically associated with erythropoiesis, with phospholipase C (PLC) and phosphatidylinositol 3-kinase (PI3K) as key players that are becoming increasingly important as new promising therapeutic targets. Additionally, Roxadustat, a new erythropoiesis stimulating agent targeting hypoxia inducible factor (HIF), is under clinical development. Here, we review the role and function of the above-mentioned signaling pathways, and we describe the state of the art and new perspectives of erythropoiesis regulation in MDS and β-thalassemia.

scholarly journals miR-8b is involved in brain and eye regeneration of Dugesia japonica in head regeneration

Biology Open ◽  
2021 ◽  
Vol 10 (6) ◽  
Author(s):  
Hongjin Liu ◽  
Qian Song ◽  
Hui Zhen ◽  
Hongkuan Deng ◽  
Bosheng Zhao ◽  
...  
Keyword(s):  
Regulate Gene Expression ◽  
Planarian Regeneration ◽  
Evolutionarily Conserved ◽  
Proliferation And Differentiation ◽  
Important Regulator ◽  
Dugesia Japonica ◽  
Non Coding Rnas ◽  
Available Information ◽  
Head Regeneration

ABSTRACT MicroRNAs (miRNAs) are a class of evolutionarily conserved small non-coding RNAs that regulate gene expression at the translation level in cell growth, proliferation and differentiation. In addition, some types of miRNAs have been proven to be key modulators of both CNS development and plasticity, such as let-7, miR-9 and miR-124. In this research, we found miR-8b acts as an important regulator involved in brain and eyespot regeneration in Dugesia japonica. miR-8b was highly conserved among species and was abundantly expressed in central nervous system. Here, we detected the expression dynamics of miR-8b by qPCR during the head regeneration of D. japonica. Knockdown miR-8b by anti-MIRs method caused severe defects of eyes and CNS. Our study revealed the evolutionary conserved role of miR-8b in the planarian regeneration process, and further provided more research ideas and available information for planarian miRNAs.

scholarly journals Collagen Hydrolysate Gly-Pro-Hyp on Osteoblastic Proliferation and Differentiation of MC3T3-E1 Cells

2017 ◽  
Vol 1 (3) ◽  
Author(s):  
Geng Min
Keyword(s):  
Alkaline Phosphatase ◽  
Transcription Factor ◽  
Bone Formation ◽  
Osteoblast Differentiation ◽  
Control Medium ◽  
Collagen Hydrolysate ◽  
Alp Activity ◽  
Proliferation And Differentiation ◽  
Related Transcription Factor ◽  
Set Up

Objectives: Bone formation and bone resorption continuouslyoccur in bone tissue to prevent the accumulation of old bone, thisbeing called bone remodeling. Osteoblasts especially play a crucialrole in bone formation through the differentiation and proliferation.Therefore, in this study, we investigated the effects of collagenhydrolysate Glycine-Prolyl-Hydroxyproline (Gly-Pro-Hyp) onosteoblastic proliferation and differentiation in MC3T3-E1 cells.Methods: Four groups including control, Gly-Pro-Hyp 20 μM,100 μM, 500 μM groups were set up in this study. Cells werecultured with blank control medium or Gly-Pro-Hyp of thedifferent dosages for 24 h. CCK 8 assay was analyzed cellproliferation. Assay of Alkaline phosphatase (ALP)activity wasanalyzed osteoblast differentiation. The expression levels of ALP,Col 1, Runx 2 and Osterix in MC3T3-E1 cells were measured byWestern blot. Results: The results indicated the treatment of Gly-Pro-Hyp promoted the proliferation of MC3T3-E1 cells andimproved ALP activity. In addition, cells treated with Gly-Pro-Hypsignificantly upregulated protein expression of ALP, type 1collagen, runt-related transcription factor 2 and osterix.Conclusion: The results demonstrate that Gly-Pro-Hyp promotedifferentiation inducement and proliferation of MC3T3-E1 cells,therefore may help to elucidate the transcriptional mechanism ofbone formation and possibly lead to the development of bone-forming drugs.

scholarly journals Insulin increases the density of potassium channels in white adipocytes: possible role in adipogenesis

2002 ◽  
Vol 174 (2) ◽  
pp. 299-307 ◽  
Author(s):  
MP Ramirez-Ponce ◽  
JC Mateos ◽  
JA Bellido
Keyword(s):  
Potassium Channels ◽  
Normal Growth ◽  
Potassium Currents ◽  
High Concentration ◽  
Differentiated Cells ◽  
White Adipocytes ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Inhibition Of Differentiation

We studied the potassium currents in white adipocytes obtained by culturing preadipocytes from rat epididymal tissue, both with insulin (WA(i)) and without insulin (WA(o)), in order to test the role of insulin in the development of voltage-gated potassium channels (K(v)) during adipogenesis. Occasionally, very small potassium currents (I(K,V)) were present in preadipocytes; however these currents were measured in all differentiated cells (adipocytes). WA(i) exhibited greater macroscopic potassium currents than WA(o) with no apparent differences in kinetics or voltage dependence. The current density (pA/ micro m(2)) calculated in WA(i) was higher than in WA(o). Currents were blocked by millimolar concentrations of tetrethylamonium (TEA). The effect of insulin on adipogenesis, both with and without TEA, was analysed. Four days without insulin and three days with insulin were necessary to increase the total number of cells in culture by 2.5-fold. Insulin increased the number of differentiated cells by 73.5%. Cell proliferation and differentiation were inhibited by TEA. Proliferation was affected only by high concentration of TEA. Inhibition of differentiation was dose dependent, with the concentration necessary for half-block similar to the IC(50) values to block potassium channels. These results suggest that insulin increases the density of K(v) and that these channels may be necessary for the normal growth of white adipocytes in culture.

scholarly journals Surface Modification of Porous Titanium with Microarc Oxidation and Its Effects on Osteogenesis ActivityIn Vitro

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Qi Wang ◽  
Mengqi Cheng ◽  
Guo He ◽  
Xianlong Zhang
Keyword(s):  
De Novo ◽  
Early Stage ◽  
Microarc Oxidation ◽  
Porous Metal ◽  
Collagen Type I ◽  
Porous Titanium ◽  
Type I ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation

Microarc oxidation (MAO) is a method about surface treatment that can provide nanoporous pits and thick oxide layers. A kind of porous metal-entangled titanium (Ti) wire material was treated with MAO process, resulting in a homogeneous rough TiO2layer, which helped facilitate MG-63 cell growth, cell viability, early cell differentiation, and cell mineralizationin vitro. In addition, the MAO-treated Ti surfaces could promote the proliferation of MG-63 cells without sacrificing differentiationin vitro, which would benefitde novobone formation around MAO-treated titanium at the early stage. The transcription levels of the extracellular matrix genes of osterix (OSX), collagen type I (Col I), bone sialoprotein (BSP), alkaline phosphatase (ALP), osteocalcin (OC) and osteopontin (OPN) and their protein expression levels were measured, suggesting that the cocultured cells with MAO titanium maintained the osteoblastic phenotype and that the MAO-treated titanium surface greatly stimulated osteoblast cell proliferation and differentiation compared to the untreated titanium. In conclusion, MAO technique can improve the surface of titanium and can contribute to the osseointegration process.

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