In vitro differentiation of neuronal precursor cells from amphibian late gastrulae: morphological, immunocytochemical studies, biosynthesis, accumulation and uptake of neurotransmitters

Development ◽  
1985 ◽  
Vol 86 (1) ◽  
pp. 71-87
Author(s):  
Anne-Marie Duprat ◽  
Paulette Kan ◽  
Françoise Foulquier ◽  
Michel Weber
Keyword(s):  
Ambystoma Mexicanum ◽  
Acetylcholine Synthesis ◽  
Toxin Binding ◽  
Neuronal Precursor Cells ◽  
Neuronal Precursors ◽  
Neurofilament Polypeptides ◽  
Pleurodeles Waltl ◽  
High Degree ◽  
Neurula Stage

Neuronal differentiation has been studied in dissociated cell cultures from early neurulae of Pleurodeles waltl and Ambystoma mexicanum. Cocultures were prepared from the neural primordium and underlying chordamesoderm. NP and NF cultures were prepared from isolated neural plate and neural folds, respectively. Neuronal precursors in NP and NF cultures had distinctive aggregation properties already evident after 1–2 days in culture. After 10–15 days, mature neurones and synapses were observed by electron microscopy in the three culture types. The expression of neurofilament polypeptides and tetanus-toxin-binding sites was also present in these cultures. A small percentage of neurones contained cytochemically detectable catecholamine. Many neurones took up tritiated dopamine with a high affinity. Quantitative measurement of [3H]acetylcholine synthesis and storage from [3H]choline were negative at the early neurula stage and in 5 to 15-day-old NF cultures, and remained low in 5 to 15-day-old NP cultures. Acetylcholine production in cocultures increased linearly with time and was always much higher than in NP cultures. These results suggest that, at the early neurula stage, some neuronal precursors have acquired the capacity to express a high degree of morphological and biochemical differentiation even in the absence of further chordamesoderm influence. However, the chordamesodermal cells in the cultures increased acetylcholine synthesis.

Neural induction: embryonic determination elicits full expression of specific neuronal traits

Development ◽  
1985 ◽  
Vol 89 (Supplement) ◽  
pp. 167-183
Author(s):  
A. M. Duprat ◽  
P. Kan ◽  
L. Gualandris ◽  
F. Foulquier ◽  
J. Marty ◽  
...  
Keyword(s):  
Neural Induction ◽  
Morphological Differentiation ◽  
Target Cells ◽  
Adhesive Properties ◽  
Neuronal Markers ◽  
Toxin Binding ◽  
Neurofilament Polypeptides ◽  
Pleurodeles Waltl ◽  
High Degree

In Pleurodeles waltl, the early neuronal differentiation of precursor cells from late gastrula stage has been studied by culture in vitro from either isolated neural plate (NP) or isolated neural fold (NF). The aim of this study was to delineate the information acquired by ectodermal target cells during neural induction. By culturing these cells in vitro either with or without the underlying chordamesoderm, we showed that in the absence of chordamesodermal influence such NP or NF cells exhibited a high degree of biochemical and morphological differentiation as revealed by the synthesis and the storage of neurotransmitters, the activity of specific enzymes, as well as by the expression of neuronal markers: specific changes in cell surface carbohydrates, tetanus toxin binding sites and neurofilament polypeptides. Remarkable changes in the cell adhesive properties were the first events observed in the different central (NP) and peripheral (NF) types. In cocultures the chordamesodermal cells exert a beneficial influence on this differentiation, specially increasing acetylcholine synthesis. There are some differences between central (NP) or peripheral (NF) neuroblast response to this further notochord or mesodermal influence.

Peptidergic properties expressed in vitro by embryonic neuroblasts after neural induction

Development ◽  
1989 ◽  
Vol 105 (3) ◽  
pp. 529-540
Author(s):  
F. Pituello ◽  
P. Deruntz ◽  
L. Pradayrol ◽  
A.M. Duprat
Keyword(s):  
Culture Medium ◽  
In Vitro Model ◽  
Neural Induction ◽  
Defined Medium ◽  
Neuronal Precursor Cells ◽  
Cell Extracts ◽  
Neuronal Precursors ◽  
Immunocytochemical Techniques ◽  
Vitro Model

As an immediate consequence of neural induction, some neuroectodermal cells acquire the ability to develop a number of characteristic neuronal features, without requiring any subsequent embryonic cues (Duprat et al. 1987). Thus, adrenergic, cholinergic and gabaergic traits are expressed in cultures of neural fold and neural plate isolated from amphibian embryos immediately after induction and grown in a defined medium. The aim of the present study was to determine, using the same in vitro model, their abilities to develop peptidergic phenotypes. Using immunocytochemical techniques, we show that substance P-, enkephalin- (leu-enkephalin, metenkephalin), and somatostatin- like immunoreactivities are expressed in subpopulations of neurones grown in vitro, whereas VIP (vasoactive intestinal polypeptide) is not detected under the same conditions. The appearance and development of the somatostatinergic phenotype has been quantified by RIA both in cell extracts and in the culture medium. Somatostatin-like immunoreactivity (SLI) undetectable at the late gastrula stage, can be measured in cells after 4 days of culture and continues to increase over the next 10 days. In culture medium, SLI is present at a constant level from day 4 up to day 14. These data reveal that some neuronal precursor cells acquire, during neural induction, the potentiality to biosynthesize, store and release neuropeptides. Furthermore, the expression of these peptidergic phenotypes in distinct subpopulations of neurones suggests that certain neuronal precursors become committed to different metabolic pathways at the earliest steps of neurogenesis.

scholarly journals Thyroid hormones promote cell differentiation and up-regulate the expression of the seladin-1 gene in in vitro models of human neuronal precursors

2008 ◽  
Vol 197 (2) ◽  
pp. 437-446 ◽  
Author(s):  
S Benvenuti ◽  
P Luciani ◽  
I Cellai ◽  
C Deledda ◽  
S Baglioni ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Cell Differentiation ◽  
Thyroid Hormones ◽  
Brain Regions ◽  
Developing Brain ◽  
Neuronal Marker ◽  
Neuronal Precursor Cells ◽  
Neuronal Precursors

Thyroid hormones (TH) play an important role in the development of human brain, by regulating the expression of specific genes. Selective Alzheimer's disease indicator-1 (seladin-1) is a recently discovered gene with neuroprotective properties, which has been found to be down-regulated in brain regions affected by Alzheimer's disease. Seladin-1 has anti-apoptotic properties mainly due to the inhibition of the activation of caspase 3. The aim of this study was to determine whether seladin-1 may be regarded as a new mediator of the effects of TH in the developing brain. In order to demonstrate this hypothesis, the effects of TH both on cell differentiation and on the expression of seladin-1 were assessed in two different cell models, i.e. fetal human neuroepithelial cells (FNC) and human mesenchymal stem cells (hMSC), which can be differentiated into neurons. 3,3′,5-Triiodothyronine (T3) determined different biological responses (inhibition of cell adhesion, induction of migration, and increase in the expression of the neuronal marker neurofilament-M and Na+ and Ca2+ channel functionality) in both FNC and hMSC, which express TH receptors. Then, we showed that TH significantly increase the expression levels of seladin-1, and that T3 effectively prevents camptothecin-induced apoptosis. However, in hMSC-derived neurons the expression of seladin-1 was not affected by TH. Our results demonstrated for the first time that seladin-1 is a novel TH-regulated gene in neuronal precursors. In view of its anti-apoptotic activity, it might be hypothesized that one of the functions of the increased seladin-1 levels in the developing brain may be to protect neuronal precursor cells from death.

scholarly journals Liposomes Prevent In Vitro Hemolysis Induced by Streptolysin O and Lysenin

Membranes ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 364
Author(s):  
Marcelo Ayllon ◽  
Gamid Abatchev ◽  
Andrew Bogard ◽  
Rosey Whiting ◽  
Sarah E. Hobdey ◽  
...  
Keyword(s):  
Blood Cell ◽  
Lipid Membranes ◽  
Bilayer Lipid Membranes ◽  
Prior Work ◽  
Cellular Targets ◽  
Toxin Binding ◽  
Streptolysin O ◽  
Biological Attack ◽  
Toxin Removal

The need for alternatives to antibiotics in the fight against infectious diseases has inspired scientists to focus on antivirulence factors instead of the microorganisms themselves. In this respect, prior work indicates that tiny, enclosed bilayer lipid membranes (liposomes) have the potential to compete with cellular targets for toxin binding, hence preventing their biological attack and aiding with their clearance. The effectiveness of liposomes as decoy targets depends on their availability in the host and how rapidly they are cleared from the circulation. Although liposome PEGylation may improve their circulation time, little is known about how such a modification influences their interactions with antivirulence factors. To fill this gap in knowledge, we investigated regular and long-circulating liposomes for their ability to prevent in vitro red blood cell hemolysis induced by two potent lytic toxins, lysenin and streptolysin O. Our explorations indicate that both regular and long-circulating liposomes are capable of similarly preventing lysis induced by streptolysin O. In contrast, PEGylation reduced the effectiveness against lysenin-induced hemolysis and altered binding dynamics. These results suggest that toxin removal by long-circulating liposomes is feasible, yet dependent on the particular virulence factor under scrutiny.

scholarly journals Chromatography-Independent Fractionation and Newly Identified Molecular Features of the Adzuki Bean (Vigna angularis Willd.) β-vignin Protein

2021 ◽  
Vol 22 (6) ◽  
pp. 3018
Author(s):  
Biane Philadelpho ◽  
Victória Souza ◽  
Fabiani Souza ◽  
Johnnie Santos ◽  
Fabiana Batista ◽  
...  
Keyword(s):  
Metal Binding ◽  
Cardiovascular Health ◽  
Binding Capacity ◽  
Biological Activities ◽  
Electrophoretic Analysis ◽  
Adzuki Bean ◽  
Molecular Features ◽  
Health Promoting ◽  
High Degree

Adzuki seed β-vignin, a vicilin-like globulin, has proven to exert various health-promoting biological activities, notably in cardiovascular health. A simple scalable enrichment procedure of this protein for further nutritional and functional studies is crucial. In this study, a simplified chromatography-independent protein fractionation procedure has been optimized and described. The electrophoretic analysis showed a high degree of homogeneity of β-vignin isolate. Furthermore, the molecular features of the purified protein were investigated. The adzuki bean β-vignin was found to have a native size of 146 kDa, and the molecular weight determined was consistent with a trimeric structure. These were identified in two main polypeptide chains (masses of 56–54 kDa) that are glycosylated polypeptides with metal binding capacity, and one minor polypeptide chain with a mass 37 kDa, wherein these features are absent. The in vitro analysis showed a high degree of digestibility of the protein (92%) and potential anti-inflammatory capacity. The results lay the basis not only for further investigation of the health-promoting properties of the adzuki bean β-vignin protein, but also for a possible application as nutraceutical molecule.

scholarly journals Construction of a mammalian embryo model from stem cells organized by a morphogen signalling centre

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Peng-Fei Xu ◽  
Ricardo Moraes Borges ◽  
Jonathan Fillatre ◽  
Maraysa de Oliveira-Melo ◽  
Tao Cheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Tracking ◽  
Cardiac Tissue ◽  
Embryonic Stem ◽  
Mammalian Embryo ◽  
Wide Range ◽  
Vitro Model ◽  
Neurula Stage

AbstractGenerating properly differentiated embryonic structures in vitro from pluripotent stem cells remains a challenge. Here we show that instruction of aggregates of mouse embryonic stem cells with an experimentally engineered morphogen signalling centre, that functions as an organizer, results in the development of embryo-like entities (embryoids). In situ hybridization, immunolabelling, cell tracking and transcriptomic analyses show that these embryoids form the three germ layers through a gastrulation process and that they exhibit a wide range of developmental structures, highly similar to neurula-stage mouse embryos. Embryoids are organized around an axial chordamesoderm, with a dorsal neural plate that displays histological properties similar to the murine embryo neuroepithelium and that folds into a neural tube patterned antero-posteriorly from the posterior midbrain to the tip of the tail. Lateral to the chordamesoderm, embryoids display somitic and intermediate mesoderm, with beating cardiac tissue anteriorly and formation of a vasculature network. Ventrally, embryoids differentiate a primitive gut tube, which is patterned both antero-posteriorly and dorso-ventrally. Altogether, embryoids provide an in vitro model of mammalian embryo that displays extensive development of germ layer derivatives and that promises to be a powerful tool for in vitro studies and disease modelling.

scholarly journals The Antibacterial Effects of Resin-Based Dental Sealants: A Systematic Review of In Vitro Studies

Materials ◽  
2021 ◽  
Vol 14 (2) ◽  
pp. 413
Author(s):  
Saad Saeed AlShahrani ◽  
Mana’a Saleh AlAbbas ◽  
Isadora Martini Garcia ◽  
Maha Ibrahim AlGhannam ◽  
Muath Abdulrahman AlRuwaili ◽  
...  
Keyword(s):  
Antibacterial Agents ◽  
Data Extraction ◽  
Risk Of Bias ◽  
Dental Sealants ◽  
Medium Risk ◽  
Metabolic Activities ◽  
High Degree ◽  
Phosphate Buffered Saline ◽  
Full Text Screening

This review aimed to assess the antimicrobial effects of different antibacterial agents/compounds incorporated in resin-based dental sealants. Four databases (PubMed, MEDLINE, Web of Science and Scopus) were searched. From the 8052 records retrieved, 275 records were considered eligible for full-text screening. Nineteen studies met the inclusion criteria. Data extraction and quality assessment was performed by two independent reviewers. Six of the nineteen included studies were judged to have low risk of bias, and the rest had medium risk of bias. Compounds and particles such as zinc, tin, Selenium, chitosan, chlorhexidine, fluoride and methyl methacrylate were found to be effective in reducing the colony-forming unit counts, producing inhibition zones, reducing the optical density, reducing the metabolic activities, reducing the lactic acid and polysaccharide production and neutralizing the pH when they are added to the resin-based dental sealants. In addition, some studies showed that the antibacterial effect was not significantly different after 2 weeks, 2 months and 6 months aging in distilled water or phosphate-buffered saline. In conclusion, studies have confirmed the effectiveness of adding antibacterial agents/compounds to dental sealants. However, we should consider that these results are based on laboratory studies with a high degree of heterogeneity.

Myocardial CO2 buffering: role of transmembrane transport of H+ or HCO3-ions

1976 ◽  
Vol 230 (4) ◽  
pp. 1037-1041 ◽  
Author(s):  
DR Strome ◽  
RL Clancy ◽  
NC Gonzalez
Keyword(s):  
Small Volume ◽  
Coronary Circulation ◽  
Myocardial Cell ◽  
Ringer Solution ◽  
Red Cells ◽  
Transmembrane Transport ◽  
High Degree

Isolated rabbit hearts were perfused with rabbit red cells suspended in Ringer solution. A small volume of perfusate was recirculated for 10 min at Pco2 of 33.4 +/- 0.9 or 150.8 +/- 7.5 mmHg. Hypercapnia resulted in an increase in perfusate HCO3- concentration that was smaller than that observed when isolated perfusate was equilibrated in vitro with the same CO2 tensions (delta HCO-3e = 1.6 mM, P less than 0.01). This difference is consistent with a net movement of HCO3- into or H+ out of the mycardial cell, and cannot be accounted for by dilution of HCO3- in the myocardial interstitium. Recirculation of perfusate through the coronary circulation at normal Pco2 for two consecutive 10-min periods was not followed by changes in perfusate HCO3- concentration. A high degree of correlation (r = 0.81) was observed between intracellular HCO-3e concentration and the corresponding delta HCO-3e in individual experiments. The results suggest that transmembrane exchange of H+ or HCO3- is a buffer mechanism for CO2 in the myocardial cell.

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