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.

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.

Ultrastructural Study of a differentiating human colon carcinoma cell line

1990 ◽  
Vol 48 (3) ◽  
pp. 208-209
Author(s):  
Sylvie Polak-Charcon ◽  
Mehrdad Hekmati ◽  
Yehuda Ben Shaul
Keyword(s):  
Cell Line ◽  
Morphological Changes ◽  
Secretory Granules ◽  
Human Colon ◽  
Cell Types ◽  
Culture Conditions ◽  
Morphological Evidence ◽  
Human Colon Carcinoma Cell ◽  
Colon Cell

The epithelium of normal human colon mucosa “in vivo” exhibits a gradual pattern of differentiation as undifferentiated stem cells from the base of the crypt of “lieberkuhn” rapidly divide, differentiate and migrate toward the free surface. The major differentiated cell type of the intestine observed are: absorptive cells displaying brush border, goblet cells containing mucous granules, Paneth and endocrine cells containing dense secretory granules. These different cell types are also found in the intestine of the 13-14 week old embryo.We present here morphological evidence showing that HT29, an adenocarcinoma of the human colon cell line, can differentiate into various cell types by changing the growth and culture conditions and mimic morphological changes found during development of the intestine in the human embryo.HT29 cells grown in tissue-culture dishes in DMEM and 10% FCS form at late confluence a multilayer of morphologically undifferentiated cell culture covered with irregular microvilli, and devoid of tight junctions (Figs 1-3).

scholarly journals The genetics of catalase in Drosophila melanogaster: isolation and characterization of acatalasemic mutants.

Genetics ◽  
1989 ◽  
Vol 122 (3) ◽  
pp. 643-652 ◽  
Author(s):  
W J Mackay ◽  
G C Bewley
Keyword(s):  
Drosophila Melanogaster ◽  
Free Radical ◽  
Catalase Activity ◽  
Oxygen Toxicity ◽  
Threshold Effect ◽  
Culture Conditions ◽  
Free Radical Damage ◽  
Isolation And Characterization ◽  
Standard Culture ◽  
Hypomorphic Alleles

Abstract Activated oxygen species have been demonstrated to be the important agents in oxygen toxicity by disrupting the structural and functional integrity of cells through lipid peroxidation events, DNA damage and protein inactivation. The biological consequences of free radical damage have long been hypothesized to be a causal agent in many aging-related diseases. Catalase (H2O2:H2O2 oxidoreductase; EC 1.15.1.1) is one of several enzymes involved in the scavenging of oxygen free radicals and free radical derivatives. The structural gene for catalase in Drosophila melanogaster has been localized to region 75D1-76A on chromosome 3L by dosage responses to segmental aneuploidy. This study reports the isolation of a stable deficiency, Df(3L)CatDH104(75C1-2;75F1), that uncovers the catalase locus and the subsequent isolation of six acatalasemic mutants. All catalase mutants are viable under standard culture conditions and recessive lethal mutations within the 75Cl-F1 interval have been shown not to affect catalase activity. Two catalase mutations are amorphic while four are hypomorphic alleles of the Cat+ locus. The lack of intergenic complementation between the six catalase mutations strongly suggests that there is only one functional gene in Drosophila. One acatalesemic mutation was mapped to position 3-47.0 which resides within the catalase dosage sensitive region. While complete loss of catalase activity confers a severe viability effect, residual levels are sufficient to restore viability to wild type levels. These results suggest a threshold effect for viability and offer an explanation for the general lack of phenotypic effects associated with the known mammalian acatalasemics.

scholarly journals Development of a potent embryonic chick lens model for studying congenital cataracts in vivo

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Zhen Li ◽  
Sumin Gu ◽  
Yumeng Quan ◽  
Kulandaiappan Varadaraj ◽  
Jean X. Jiang
Keyword(s):  
Light Transmission ◽  
Genetic Diseases ◽  
Gene Mutations ◽  
Underlying Mechanism ◽  
Dominant Negative ◽  
Lens Model ◽  
Embryonic Chick ◽  
Congenital Cataracts ◽  
Chick Lens

AbstractCongenital cataracts are associated with gene mutations, yet the underlying mechanism remains largely unknown. Here we reported an embryonic chick lens model that closely recapitulates the process of cataract formation. We adopted dominant-negative site mutations that cause congenital cataracts, connexin, Cx50E48K, aquaporin 0, AQP0R33C, αA-crystallin, CRYAA R12C and R54C. The recombinant retroviruses containing these mutants were microinjected into the occlusive lumen of chick lenses at early embryonic development. Cx50E48K expression developed cataracts associated with disorganized nuclei and enlarged extracellular spaces. Expression of AQP0R33C resulted in cortical cataracts, enlarged extracellular spaces and distorted fiber cell organization. αA crystallin mutations distorted lens light transmission and increased crystalline protein aggregation. Together, retroviral expression of congenital mutant genes in embryonic chick lenses closely mimics characteristics of human congenital cataracts. This model will provide an effective, reliable in vivo system to investigate the development and underlying mechanism of cataracts and other genetic diseases.

scholarly journals Selecting single cell clustering parameter values using subsampling-based robustness metrics

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Ryan B. Patterson-Cross ◽  
Ariel J. Levine ◽  
Vilas Menon
Keyword(s):  
Single Cell ◽  
Optimal Parameter ◽  
Clustering Algorithms ◽  
Cell Types ◽  
Parameter Selection ◽  
Data Set ◽  
Biologically Relevant ◽  
Cell Clustering ◽  
Parameter Values ◽  
Robustness Metrics

Abstract Background Generating and analysing single-cell data has become a widespread approach to examine tissue heterogeneity, and numerous algorithms exist for clustering these datasets to identify putative cell types with shared transcriptomic signatures. However, many of these clustering workflows rely on user-tuned parameter values, tailored to each dataset, to identify a set of biologically relevant clusters. Whereas users often develop their own intuition as to the optimal range of parameters for clustering on each data set, the lack of systematic approaches to identify this range can be daunting to new users of any given workflow. In addition, an optimal parameter set does not guarantee that all clusters are equally well-resolved, given the heterogeneity in transcriptomic signatures in most biological systems. Results Here, we illustrate a subsampling-based approach (chooseR) that simultaneously guides parameter selection and characterizes cluster robustness. Through bootstrapped iterative clustering across a range of parameters, chooseR was used to select parameter values for two distinct clustering workflows (Seurat and scVI). In each case, chooseR identified parameters that produced biologically relevant clusters from both well-characterized (human PBMC) and complex (mouse spinal cord) datasets. Moreover, it provided a simple “robustness score” for each of these clusters, facilitating the assessment of cluster quality. Conclusion chooseR is a simple, conceptually understandable tool that can be used flexibly across clustering algorithms, workflows, and datasets to guide clustering parameter selection and characterize cluster robustness.

Structure of vegetative organs in essential oil rose under standard culture conditions and long-term conservation in vitro

2021 ◽  
pp. 185-190
Author(s):  
I.V. Mitrofanova ◽  
V.A. Brailko ◽  
N.P. Lesnikova-Sedoshenko ◽  
N.N. Ivanova ◽  
O.V. Mitrofanova
Keyword(s):  
Essential Oil ◽  
Culture Conditions ◽  
Vegetative Organs ◽  
Standard Culture

scholarly journals Muscle contracture and passive mechanics in cerebral palsy

2019 ◽  
Vol 126 (5) ◽  
pp. 1492-1501 ◽  
Author(s):  
Richard L. Lieber ◽  
Jan Fridén
Keyword(s):  
Extracellular Matrix ◽  
Cerebral Palsy ◽  
Satellite Cells ◽  
Neuromuscular Disorders ◽  
Light And Electron Microscopy ◽  
Cell Types ◽  
Underlying Mechanism ◽  
Muscle Stiffness ◽  
Muscle Satellite Cells ◽  
Cord Injury

Skeletal muscle contractures represent the permanent shortening of a muscle-tendon unit, resulting in loss of elasticity and, in extreme cases, joint deformation. They may result from cerebral palsy, spinal cord injury, stroke, muscular dystrophy, and other neuromuscular disorders. Contractures are the prototypic and most severe clinical presentation of increased passive mechanical muscle force in humans, often requiring surgical correction. Intraoperative experiments demonstrate that high muscle passive force is associated with sarcomeres that are abnormally stretched, although otherwise normal, with fewer sarcomeres in series. Furthermore, changes in the amount and arrangement of collagen in the extracellular matrix also increase muscle stiffness. Structural light and electron microscopy studies demonstrate that large bundles of collagen, referred to as perimysial cables, may be responsible for this increased stiffness and are regulated by interaction of a number of cell types within the extracellular matrix. Loss of muscle satellite cells may be related to changes in both sarcomeres and extracellular matrix. Future studies are required to determine the underlying mechanism for changes in muscle satellite cells and their relationship (if any) to contracture. A more complete understanding of this mechanism may lead to effective nonsurgical treatments to relieve and even prevent muscle contractures.

Experimental studies on Lecanora rupicola (L.) Zahlbr.: chemical and microscopical investigations of the mycobiont and re-synthesis stages

2006 ◽  
Vol 38 (6) ◽  
pp. 577-585 ◽  
Author(s):  
Georg BRUNAUER ◽  
Armin HAGER ◽  
Wolf Dietrich KRAUTGARTNER ◽  
Roman TÜRK ◽  
Elfie STOCKER-WÖRGÖTTER
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Sequence Data ◽  
Experimental Studies ◽  
Culture Conditions ◽  
Voucher Specimen ◽  
Dna Sequence Data ◽  
Standard Culture ◽  
Voucher Specimens ◽  
Scanning Electron

Culture experiments that trigger the axenically grown mycobionts of Lecanora rupicola to produce the polyketide chemosyndrome typical of the naturally grown lichen are reported. This chemosyndrome comprises lecanoric, haematommic and orsellinic acids, sordidone, eugenitol and atranorin, all of which were hardly produced under standard culture conditions. The only exception was arthothelin that was only present in the voucher specimen. It has been shown that almost the complete acetyl-polymalonyl-pathway leading to depsides and chromones can be induced in culture, but apparently not the xanthones. The mycobiont was also successfully re-synthesized with its original photobiont, as confirmed by Scanning Electron Microscope studies (SEM). Cultures of the resynthesised lichen biosynthesized additional satellite substances, which were not detected either in the voucher specimens or in the aposymbiontically (without the photobiont) grown mycobiont cultures. The identity of cultured mycobionts of L. rupicola was confirmed by comparing ITS-DNA-sequence data from the original lichen with publicly available (GeneBank) sequences of that species.

scholarly journals Apamin Enhances Neurite Outgrowth and Regeneration after Laceration Injury in Cortical Neurons

Toxins ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 603
Author(s):  
Hyunseong Kim ◽  
Jin Young Hong ◽  
Junseon Lee ◽  
Wan-Jin Jeon ◽  
In-Hyuk Ha
Keyword(s):  
Phospholipase A2 ◽  
Neurite Outgrowth ◽  
Cortical Neurons ◽  
Neurological Diseases ◽  
Minor Component ◽  
Underlying Mechanism ◽  
Bee Venom ◽  
Dependent Manner ◽  
Neuroprotective Effects ◽  
Wide Range

Apamin is a minor component of bee venom and is a polypeptide with 18 amino acid residues. Although apamin is considered a neurotoxic compound that blocks the potassium channel, its neuroprotective effects on neurons have been recently reported. However, there is little information about the underlying mechanism and very little is known regarding the toxicological characterization of other compounds in bee venom. Here, cultured mature cortical neurons were treated with bee venom components, including apamin, phospholipase A2, and the main component, melittin. Melittin and phospholipase A2 from bee venom caused a neurotoxic effect in dose-dependent manner, but apamin did not induce neurotoxicity in mature cortical neurons in doses of up to 10 µg/mL. Next, 1 and 10 µg/mL of apamin were applied to cultivate mature cortical neurons. Apamin accelerated neurite outgrowth and axon regeneration after laceration injury. Furthermore, apamin induced the upregulation of brain-derived neurotrophic factor and neurotrophin nerve growth factor, as well as regeneration-associated gene expression in mature cortical neurons. Due to its neurotherapeutic effects, apamin may be a promising candidate for the treatment of a wide range of neurological diseases.

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