scholarly journals Characterization of a factor that promotes neurite outgrowth: evidence linking activity to a heparan sulfate proteoglycan.

1982 ◽  
Vol 94 (3) ◽  
pp. 574-585 ◽  
Author(s):  
A D Lander ◽  
D K Fujii ◽  
D Gospodarowicz ◽  
L F Reichardt
Keyword(s):  
Endothelial Cells ◽  
Neurite Outgrowth ◽  
Sympathetic Neurons ◽  
Cesium Chloride ◽  
Cell Types ◽  
Sulfate Proteoglycan ◽  
Corneal Endothelial Cells ◽  
Active Factor ◽  
Heparin Sulfate Proteoglycan ◽  
Heparin Sulfate

Rat sympathetic neurons, plated onto extracellular matrix produced by cultured bovine corneal endothelial cells, rapidly extended neurites in the absence of nerve growth factor (NGF). The response was unaffected by antiserum to NGF. Rapid outgrowth also occurred when sympathetic neurons were plated onto polylysine-coated surfaces that had been exposed to serum-free medium conditioned by corneal endothelial cells (CMSF). A response was seen even when the neurons were cultured without serum. When plated onto a polylysine-coated dish treated with CMSF over half its surface, only the neurons on the treated half extended neurites. The active factor in CMSF was destroyed by trypsin, acid (pH 1.6), base (pH 12.7), or heating to 80 degrees C; it was stable to heating to 60 degrees C, collagenase, deoxyribonuclease, and neuraminidase. The factor elutes just after the void volume of a Sepharose 6B column. In associative cesium chloride gradients, it sediments as a peak centered at a density of 1.36-1.37, corresponding to a peak of material that can be biosynthetically labeled with [35S]sulfate or [3H]leucine. Material from this fraction was inactivated by heparinase, but not chondroitinase ABC, implying that a heparin sulfate proteoglycan is essential for the factor's activity. Inactivation by contaminants in the heparinase preparation was ruled out. Further purification indicated that the active factor may exist as an aggregate containing a heparin sulfate proteoglycan and other molecules. CMSF also promoted neurite outgrowth by other types of neurons. Furthermore, a variety of cell types were shown to produce factors similar to that in CMSF.

scholarly journals Purification of a factor that promotes neurite outgrowth: isolation of laminin and associated molecules.

1985 ◽  
Vol 101 (3) ◽  
pp. 898-913 ◽  
Author(s):  
A D Lander ◽  
D K Fujii ◽  
L F Reichardt
Keyword(s):  
Endothelial Cells ◽  
Heparan Sulfate ◽  
Neurite Outgrowth ◽  
Conditioned Medium ◽  
Culture Medium ◽  
Cell Types ◽  
Cultured Neurons ◽  
Sulfate Proteoglycan ◽  
Corneal Endothelial Cells ◽  
Laminin Binding

When culture medium, conditioned by any of several cell types, is applied to a polycationic substratum, a substance is adsorbed that causes neurons cultured on that substratum to extend processes (neurites) rapidly and profusely. We have purified the factor responsible for this effect from medium conditioned by bovine corneal endothelial cells, and have shown that it is composed of the glycoprotein laminin and two associated laminin-binding molecules: a sulfated protein known as entactin, and a large heparan sulfate proteoglycan. Of these molecules, only laminin was found to be present throughout the purification in all fractions possessing neurite outgrowth-promoting activity and absent from all fractions lacking activity. Laminin, purified from other sources, has been shown previously to promote extensive outgrowth by cultured neurons. These and other data presented here support the conclusion that laminin is responsible for the neurite outgrowth-promoting activity of the conditioned medium factor. Evidence is also presented that the association of a proteoglycan with laminin promotes efficient attachment of laminin to polycationic substrata, particularly in the presence of competing molecules.

scholarly journals Rapid Cell Type-Dependent Uptake of 0N4R TAU Monomer is not Solely Heparin Sulfate Proteoglycan Dependen

2020 ◽  
Vol 118 (3) ◽  
pp. 454a
Author(s):  
Anne S. Robinson ◽  
Daniel Oseid ◽  
Evan Wells ◽  
Liqing Song
Keyword(s):  
Sulfate Proteoglycan ◽  
Cell Type ◽  
Heparin Sulfate Proteoglycan ◽  
Heparin Sulfate

scholarly journals A comparison of the biological activities of the cell-adhesive proteins vitronectin and fibronectin

1989 ◽  
Vol 93 (4) ◽  
pp. 641-649
Author(s):  
P.A. Underwood ◽  
F.A. Bennett
Keyword(s):  
Endothelial Cells ◽  
Serum Proteins ◽  
Cell Attachment ◽  
Biological Activities ◽  
Cell Types ◽  
Corneal Endothelial Cells ◽  
Response Curves ◽  
Similar Dose ◽  
Bovine Endothelial Cells ◽  
Adhesive Proteins

The effects of vitronectin and fibronectin upon the attachment and growth of bovine corneal endothelial cells (BCE) and BHK-21 cells were compared. Similar dose-response curves for cell attachment to the substratum were obtained for both molecules and both cell types, although BCE cells exhibited a slight preference for vitronectin, and BHK cells for fibronectin. When, however, cells were plated in medium containing bovine serum stripped of fibronectin, they attached and grew normally, whereas in medium containing serum stripped of vitronectin, cells either failed to attach (BHK-21) or attached but exhibited poor cell spreading and growth. This dependence of cells upon vitronectin, rather than fibronectin, in serum for cell attachment, was shown to be due to a failure of fibronectin to coat the substratum in the presence of other serum proteins. Vitronectin was able to coat the substratum efficiently in the presence of other serum proteins. Although dependent upon vitronectin for adhesion to the substratum, bovine endothelial cells were unable to synthesize endogenous vitronectin.

scholarly journals Segregated neural explants exhibit co-oriented, asymmetric, neurite outgrowth

2019 ◽  
Author(s):  
David B. Pettigrew ◽  
Curtis B. Dobson ◽  
Lori G. Isaacson ◽  
Eric Leuthardt ◽  
Heather Lilley ◽  
...  
Keyword(s):  
Neurite Outgrowth ◽  
Sympathetic Neurons ◽  
Cell Types ◽  
Underlying Mechanism ◽  
Culture Conditions ◽  
Emergent Behavior ◽  
Embryonic Chick ◽  
Neurite Length ◽  
Cell Clustering ◽  
Standard Culture

AbstractExplants of embryonic chick sympathetic and sensory ganglia were found to exhibit asymmetric radial outgrowth of neurites under standard culture conditions with or without exogenous Nerve Growth Factor [NGF]. Opposing sides of an explant exhibited: a] differences in neurite length and, b] differences in terminal morphology. Strikingly, this asymmetry exhibited co-orientation among segregated, neighboring explants. The underlying mechanism[s] of the asymmetry and its co-orientation have not been discovered but appears to depend on cell clustering because dissociated sympathetic neurons do not exhibit co-orientation whereas re-aggregated clusters of cells do. This emergent behavior may be similar to the community effect described in other cell types. If a similar phenomenon exists in the embryo, or in maturity, it may contribute to the establishment of proper orientation of neurite outgrowth during development and/or injury-induced neuronal plasticity.

THROMBIN INTERACTION WITH CULTURED AORTIC AND CAPILLARY ENDOTHELIAL CELLS: BINDING, INTERNALIZATION, DEGRADATION AND RELEASE OF PROTEASE NEXINS

1987 ◽  
Author(s):  
N Savion ◽  
A Gamliel ◽  
N Farzame
Keyword(s):  
Molecular Weight ◽  
Endothelial Cells ◽  
Cell Surface ◽  
Serine Proteases ◽  
Vascular Endothelial Cells ◽  
Polyacrylamide Gel Electrophoresis ◽  
Vascular Endothelial Cell ◽  
Cell Types ◽  
Vascular Endothelial ◽  
Corneal Endothelial Cells

Thrombin (Th) binds specifically to confluent cultures of bovine aortic (ABAE) and brain capillary (BBC) endothelial cells. Saturation of 125I-Th binding is observed after 1 h exposure to the ligand and at an extracellular concentration of 0.5 and 1.0 µg/ml for ABAE and BBC cells, respectively. Under optimal conditions both ABAE and BBC cultures bind about 2 to 5 ng/106 cells, which represents about 20% of Th binding.to bovine corneal endothelial (BCE) cells. The cell associated 125I-Th in ABAE and BBC cells is internalized and degraded as described in BCE cells. The nature of the cell associated radioactivity is analyzed on SDS-polyacrylamide -gel electrophoresis and in ABAE and BBC cells about 30% of the I-Th appears in a complex with protease nexin I (PN I) while in BCE cells about 70% of the binding is mediated by PN I. ABAE cells possess 3 types of complexes, one which appears only on the cell surface with a molecular weight of 78 kDa, and two other complexes which appear only in the conditioned medium (CM) with molecular weights of 84 and 85 kDa. BBC and BCE cells demonstrate only one type of complex with a molecular weight of 77 kDa which appears both on the cell surface and in the CM. Preincubation of BCE cultures in the presence of Th is known to up-regulate the amount of PN I on the cell surface and in the CM, but this Th induced up-regulation effect is not observed in ABAE or BBC cells.The results described above indicate a difference between ABAE and BBC cells although both cell types growunder similar conditins and demonstrate similar morphological appearance. However, in both vascular endothelial cell types the total amount of PN I and its metabolism is relatively small compared to corneal endothelial cells. It, therefore, may indicate the lower capacity of vascular endothelial cells to control serine proteases activity at or near their cell surfaces as compared to corneal endothelial cells. This research was supported by a grant from the NationalCouncil for Research and Development, Israel and G.S.F. Munchen, Germany

Culturing Discarded Peripheral Human Corneal Endothelial Cells From the Tissues Deemed for Preloaded DMEK Transplants

Cornea ◽  
2019 ◽  
Vol 38 (9) ◽  
pp. 1175-1181 ◽  
Author(s):  
Mohit Parekh ◽  
Vito Romano ◽  
Alessandro Ruzza ◽  
Stephen B. Kaye ◽  
Diego Ponzin ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Corneal Endothelial Cells ◽  
Human Corneal Endothelial Cells

Morphilogy and Ultrastructure of in Vitro Cultures of Aortic Endothelial Cells Interacting with Antibodies

1979 ◽  
Author(s):  
S. Korach ◽  
D. Ngo
Keyword(s):  
Endothelial Cells ◽  
Cell Surface ◽  
Vascular Endothelial Cells ◽  
Intercellular Junctions ◽  
Cell Types ◽  
Endothelial Damage ◽  
Antibody Concentration ◽  
High Yields ◽  
Scanning Em

Adult pig aortas, sectioned longitudinally, were incubated in 0.1% collagenase-PBS (15 mn, 37°C). Gentle scraping of the lumenal surface resulted in high yields (3-4 x 106 cell/aorta) of viable endothelial cells, essentially devoid of other cell types by morphological and immunochemical (F VIII-antigen) criteria. Confluent monolayers were incubated for various times (5 mn to 1 wk) with decomplemented rabbit antisera raised against pig endothelial cells. Changes in cell morphology appeared to depend on antibody concentration rather than on duration of contact with antiserum. High concentrations of antiserum (5 to 20%) led to cytoplasmic shredding, bulging of cells and extensive vacuolization, whereas at lower concentrations, cells appeared almost normal. Transmission EM studies by the indirect immunoperoxydase method showed antibodies reacting with unfixed cells to be distributed all over the upper cell surface, in the outer parts of intercellular junctions, and within numerous pinocytotic vesicles. Much weaker reactions could also be seen at the lower cell surface. When viewed under the Scanning EM, antiserum-treated endothelial cells also disclosed antibody concentration-dependent bulging and release of cells from their substrate. In vitro studies of gradual modifications of vascular endothelial cells acted upon by antibodies should provide a better understanding of the structural and biochemical processes underlying endothelial damage and detachment.

scholarly journals The Cellular Choreography of Osteoblast Angiotropism in Bone Development and Homeostasis

2021 ◽  
Vol 22 (14) ◽  
pp. 7253
Author(s):  
Georgiana Neag ◽  
Melissa Finlay ◽  
Amy J. Naylor
Keyword(s):  
Endothelial Cells ◽  
Bone Formation ◽  
Drug Targets ◽  
Bone Development ◽  
Cell Types ◽  
Regulatory Relationship ◽  
Sequencing Technologies ◽  
Resolution Imaging ◽  
Molecular Relationships ◽  
Form Type

Interaction between endothelial cells and osteoblasts is essential for bone development and homeostasis. This process is mediated in large part by osteoblast angiotropism, the migration of osteoblasts alongside blood vessels, which is crucial for the homing of osteoblasts to sites of bone formation during embryogenesis and in mature bones during remodeling and repair. Specialized bone endothelial cells that form “type H” capillaries have emerged as key interaction partners of osteoblasts, regulating osteoblast differentiation and maturation and ensuring their migration towards newly forming trabecular bone areas. Recent revolutions in high-resolution imaging methodologies for bone as well as single cell and RNA sequencing technologies have enabled the identification of some of the signaling pathways and molecular interactions that underpin this regulatory relationship. Similarly, the intercellular cross talk between endothelial cells and entombed osteocytes that is essential for bone formation, repair, and maintenance are beginning to be uncovered. This is a relatively new area of research that has, until recently, been hampered by a lack of appropriate analysis tools. Now that these tools are available, greater understanding of the molecular relationships between these key cell types is expected to facilitate identification of new drug targets for diseases of bone formation and remodeling.

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