scholarly journals Biosynthesis of anandamide and related acylethanolamides in mouse J774 macrophages and N18 neuroblastoma cells

1996 ◽  
Vol 316 (3) ◽  
pp. 977-984 ◽  
Author(s):  
Vincenzo Di MARZO ◽  
Luciano De PETROCELLIS ◽  
Nunzio SEPE ◽  
Anna BUONO
Keyword(s):  
Cell Function ◽  
Cannabinoid Receptor ◽  
Neuroblastoma Cells ◽  
Neuronal Cell ◽  
Cell Types ◽  
Endogenous Ligand ◽  
Alternative Pathways ◽  
Central Neurons ◽  
The Central Nervous System ◽  
Hydrolysis Of

Anandamide (arachidonoylethanolamide, AnNH) has been recently proposed as the endogenous ligand at the brain cannabinoid receptor CB1. Two alternative pathways have been suggested for the biosynthesis of this putative mediator in the central nervous system. Here we present data (1) substantiating further the mechanism by which AnNH is produced by phospholipase D (PLD)-catalysed hydrolysis of N-arachidonoylphosphatidylethanolamine in mouse neuroblastoma N18TG2 cells, and (2) suggesting for the first time that AnNH is biosynthesized via the same mechanism in a non-neuronal cell line, mouse J774 macrophages, together with other acylethanolamides and is possibly involved in the control of the immune/inflammatory response. Lipids from both neuroblastoma cells and J774 macrophages were shown to contain a family of N-acylphosphatidylethanolamines (N-aPEs), including the possible precursor of AnNH, N-arachidonoyl-PE. Treatment with exogenous PLD, but not with exogenous phospholipase A2 and ethanolamine, resulted in the production of a series of acylethanolamides (AEs), including AnNH, from both cell types. The formation of AEs was accompanied by a decrease in the levels of the corresponding N–aPEs. Enzymically active homogenates from either neuroblastoma cells or J774 macrophages were shown to convert synthetic N-[3H]arachidonoyl-PE into [3H]AnNH, thus suggesting that in both cells an enzyme is present which is capable of catalysing the hydrolysis of N–aPE(s) to the corresponding AE(s). Finally, as previously shown in central neurons, on stimulation with ionomycin, J774 macrophages also produced a mixture of AEs including AnNH and palmitoylethanolamide, which has been proposed as the preferential endogenous ligand at the peripheral cannabinoid receptor CB2 and, consequently, as a possible down-modulator of mast cells. On the basis of this as well as previous findings it is now possible to hypothesize for AnNH and palmitoylethanolamide, co-synthesized by macrophages, a role as peripheral mediators with multiple actions on blood cell function.

Expression of protocadherin 11Yb (PCDH11Yb) in seminal germ cells is correlated with fertility status in men

2017 ◽  
Vol 29 (11) ◽  
pp. 2100 ◽  
Author(s):  
Thottathil R. Anilkumar ◽  
Anandavalli N. Devi ◽  
Sathy M. Pillai ◽  
Krishnapillai Jayakrishnan ◽  
Oommen V. Oommen ◽  
...  
Keyword(s):  
Germ Cells ◽  
Neuronal Cell ◽  
Cell Types ◽  
Wnt Signalling ◽  
Retinoic Acid Signalling ◽  
Fertility Status ◽  
Neuronal Cell Differentiation ◽  
The Central Nervous System ◽  
Differentiation And Proliferation ◽  
Canonical Wnt

Protocadherin 11 Y-linked (PCDH11Y), a member of the cadherin superfamily, is predominantly expressed in the central nervous system, is encoded by the Yp11.2 locus and exists in three isoforms: 11Ya, 11Yb and 11Yc. PCDH11Y is upregulated by retinoic acid signalling and is essential for spermatogonial differentiation and initiation of meiosis. PCDH11Y mediates Wnt signalling, which plays a crucial role in the differentiation of various cell types. PCDH11Y has been implicated in neuronal cell differentiation and proliferation, but its association with spermatogenesis has not yet been addressed. Hence, in order to address the possible role of PCDH11Y in relation to spermatogenesis, the expression analysis of PCDH11Y in the seminal germ cells of fertile and infertile males were carried out employing RT-PCR, western blotting and immunofluorescence analysis. In the present study, PCDH11Yb, but not PCDH11Ya or PCDH11Yc, was expressed in germ cells isolated from the semen of all 13 men with proven fertility. However, in several subjects from various infertility classes, there was complete absence or a significant reduction in the expression of PCDH11Yb. PCDH11Y exhibited prominent localisation on the head and midpiece region of spermatozoa from fertile men, whereas spermatozoa from infertile subjects had either weak or abnormal localisation patterns for PCDH11Y. In addition, downregulation of canonical Wnt signalling was correlated with defective expression of PCDH11Y in spermatozoa of infertile men, as evidenced by downregulation of the Wnt targets C-Myc and C-Jun. In conclusion, expression levels of PCDH11Yb in germ cells in the semen were correlated with the fertility status of men.

A Novel Receptor for N-Arachidonoyl Dopamine Mediates Intracellular Signaling Events in Human T-Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3861-3861
Author(s):  
Florin Tuluc ◽  
Rahil T. Rahim ◽  
Satya P. Kunapuli
Keyword(s):  
T Cells ◽  
Intracellular Calcium ◽  
Intracellular Signaling ◽  
Cannabinoid Receptors ◽  
Cell Function ◽  
Cannabinoid Receptor ◽  
Calcium Influx ◽  
Cell Types ◽  
Calcium Mobilization ◽  
Human T Cells

Abstract N-arachidonoyl dopamine (NADA) is a relatively new member of the expanding family of endocannabinoids. NADA is also able to activate the transient receptor potential (TRP) channel V1, which is permeable for calcium and sodium. TRPV1 is the receptor for capsaicin and is present in dorsal horn neurons but it is also functional in other cell types. In a recent report NADA has been shown to effectively inhibit the production of IL-2 and TNFα in human T cells by negatively regulating the activation of several transcription factors, such as NFκB, NF-AT, and AP-1, strongly suggesting that NADA may play important roles in regulating T cell function. However, the signaling mechanisms triggered by NADA are poorly studied in lymphocytes. In the present study we show that NADA activates specific intracellular signaling pathways in isolated human T cells and we tested the hypothesis that these effects are mediated by a cannabinoid receptor or/and the TRPV1 channel. NADA (1 to 100 μM) induced intracellular calcium increase in fura 2-loaded human T cells in a dose-dependent manner. The effect of NADA on free cytosolic calcium is biphasic: it initially triggers mobilization of calcium from the intracellular stores followed by a strong calcium influx lasting more than 15 minutes. In addition, we show that NADA can trigger robust phosphorylation of protein kinase C𝛉 as well as the phosphorylation of p44/42 MAP kinase with a time course similar to that observed for intracellular calcium mobilization. Arachidonic acid elicited no calcium increase in isolated T cells, although it caused calcium increase in isolated human granulocytes. Moreover, phenidone (100 μM), a 5-lipoxygenase inhibitor, did not inhibit the calcium response to NADA, demonstrating that the effect of NADA on T cells is not due to generation of arachidonic acid metabolites. Cannabinoid receptors have been reported to inhibit rather than activate calcium channels. However, the CB2 cannabinoid receptor has been linked to calcium mobilization by activation of PLC through the βγ subunit of Gi proteins. We tested the effects of arachidonyl cyclopropylamide, a potent CB1 agonist, and JWH015, a potent CB2 agonist, on isolated human T cells to determine if activation of cannabinoid receptors can explain NADA-induced calcium mobilization. Neither of these two cannabinoid agonists altered intracellular calcium levels when tested at concentrations up to 10 μM. Moreover, AM251 (10 μM), an antagonist at cannabinoid receptors, did not inhibit the effect of NADA on calcium increase, further supporting the view that the effect of NADA on intracellular calcium levels in T cells is not mediated by cannabinoid receptors. Although NADA has been shown in other cell types to cause calcium influx through TRPV1 channels, capsaicin, a classical TRPV1 agonist, failed to alter the intracellular calcium levels in T cells. In addition, capsazepine (100 μM), a TRPV1 antagonist, did not inhibit the response to NADA, strongly suggesting that TRPV1 does not mediate the effect of NADA on calcium influx in human T cells. Our data support the view that NADA may function as an important endogenous modulator on T cell function, activating a novel receptor that has yet to be identified and which is distinct from either cannabinoid receptors or the TRPV1 channel.

scholarly journals Laminin N-terminus α31 protein distribution in adult human tissues

2020 ◽  
Author(s):  
Lee D. Troughton ◽  
Raphael Reuten ◽  
Conor J. Sugden ◽  
Kevin J. Hamill
Keyword(s):  
Cell Function ◽  
Basal Layer ◽  
Alveolar Epithelium ◽  
Cell Types ◽  
Epithelial Tissue ◽  
Protein Distribution ◽  
N Terminus ◽  
The Central Nervous System ◽  
Kidney Liver ◽  
Different Cell Types

AbstractLaminin N terminus α31 (LaNt α31) is a netrin-like protein derived from alternative splicing of the laminin α3 gene. Although LaNt α31 has been demonstrated to influence corneal and skin epithelial cell function, its expression has not been investigated beyond these tissues. In this study, we used immunohistochemistry to characterise the distribution of this protein in a wide-array of human tissue sections in comparison to laminin α3. These data revealed widespread LaNt α31 expression. In epithelial tissue, LaNt α31 was present in the basal layer of the epidermis, throughout the epithelium of the digestive tract, and much of the epithelium of the reproductive system. LaNt α31 was also found throughout the vasculature of most tissues, with enrichment in reticular-like fibres in the extracellular matrix surrounding large vessels. A similar matrix pattern was observed around the terminal ducts in the breast and around the alveolar epithelium in the lung, where basement membrane staining was also evident. Specific enrichment of LaNt α31 was identified in sub-populations of cells of the kidney, liver, pancreas, and spleen, with variations in intensity between different cell types in the collecting ducts and glomeruli of the kidney being of particular note. Intriguingly, LaNt α31 immunoreactivity was also evident in neurons of the central nervous system, in the cerebellum, cerebral cortex, and spinal cord. Together these findings suggest that LaNt α31 may be functionally relevant in a wider range of tissue contexts than previously thought, and provides a valuable basis for investigation into this interesting protein.

scholarly journals Laminin N-terminus α31 protein distribution in adult human tissues

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0239889
Author(s):  
Lee D. Troughton ◽  
Raphael Reuten ◽  
Conor J. Sugden ◽  
Kevin J. Hamill
Keyword(s):  
Cell Function ◽  
Basal Layer ◽  
Alveolar Epithelium ◽  
Cell Types ◽  
Epithelial Tissue ◽  
Protein Distribution ◽  
N Terminus ◽  
The Central Nervous System ◽  
Kidney Liver ◽  
Different Cell Types

Laminin N-terminus α31 (LaNt α31) is a netrin-like protein derived from alternative splicing of the laminin α3 gene. Although LaNt α31 has been demonstrated to influence corneal and skin epithelial cell function, its expression has not been investigated beyond these tissues. In this study, we used immunohistochemistry to characterise the distribution of this protein in a wide-array of human tissue sections in comparison to laminin α3. The data revealed widespread LaNt α31 expression. In epithelial tissue, LaNt α31 was present in the basal layer of the epidermis, throughout the epithelium of the digestive tract, and in much of the epithelium of the reproductive system. LaNt α31 was also found throughout the vasculature of most tissues, with enrichment in reticular-like fibres in the extracellular matrix surrounding large vessels. A similar matrix pattern was observed around the terminal ducts in the breast and around the alveolar epithelium in the lung, where basement membrane staining was also evident. Specific enrichment of LaNt α31 was identified in sub-populations of cells of the kidney, liver, pancreas, and spleen, with variations in intensity between different cell types in the collecting ducts and glomeruli of the kidney. Intriguingly, LaNt α31 immunoreactivity was also evident in neurons of the central nervous system, in the cerebellum, cerebral cortex, and spinal cord. Together these findings suggest that LaNt α31 may be functionally relevant in a wider range of tissue contexts than previously anticipated, and the data provides a valuable basis for investigation into this interesting protein.

Visualization of co-localization in Aβ42-administered neuroblastoma cells reveals lysosome damage and autophagosome accumulation related to cell death

2011 ◽  
Vol 441 (2) ◽  
pp. 579-590 ◽  
Author(s):  
Violetta Soura ◽  
Maris Stewart-Parker ◽  
Thomas L. Williams ◽  
Arjuna Ratnayaka ◽  
Joe Atherton ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Function ◽  
Neuroblastoma Cells ◽  
Amyloid Β ◽  
Neuronal Cell ◽  
Immunogold Electron Microscopy ◽  
Amyloid Β Peptide ◽  
Aβ Oligomers ◽  
Oligomeric Form ◽  
Endosomal System

Aβ42 [amyloid-β peptide-(1–42)] plays a central role in Alzheimer's disease and is known to have a detrimental effect on neuronal cell function and survival when assembled into an oligomeric form. In the present study we show that administration of freshly prepared Aβ42 oligomers to a neuroblastoma (SH-SY5Y) cell line results in a reduction in survival, and that Aβ42 enters the cells prior to cell death. Immunoconfocal and immunogold electron microscopy reveal the path of the Aβ42 with time through the endosomal system and shows that it accumulates in lysosomes. A 24 h incubation with Aβ results in cells that have damaged lysosomes showing signs of enzyme leakage, accumulate autophagic vacuoles and exhibit severely disrupted nuclei. Endogenous Aβ is evident in the cells and the results of the present study suggest that the addition of Aβ oligomers disrupts a crucial balance in Aβ conformation and concentration inside neuronal cells, resulting in catastrophic effects on cellular function and, ultimately, in cell death.

scholarly journals Sorting Out Sorting Nexins Functions in the Nervous System in Health and Disease

2021 ◽  
Author(s):  
Neide Vieira ◽  
Teresa Rito ◽  
Margarida Correia-Neves ◽  
Nuno Sousa
Keyword(s):  
Nervous System ◽  
Protein Trafficking ◽  
Cell Function ◽  
Neuronal Cell ◽  
Protein Family ◽  
Sorting Nexins ◽  
Genome Wide ◽  
Fundamental Process ◽  
The Central Nervous System ◽  
Health And Disease

AbstractEndocytosis is a fundamental process that controls protein/lipid composition of the plasma membrane, thereby shaping cellular metabolism, sensing, adhesion, signaling, and nutrient uptake. Endocytosis is essential for the cell to adapt to its surrounding environment, and a tight regulation of the endocytic mechanisms is required to maintain cell function and survival. This is particularly significant in the central nervous system (CNS), where composition of neuronal cell surface is crucial for synaptic functioning. In fact, distinct pathologies of the CNS are tightly linked to abnormal endolysosomal function, and several genome wide association analysis (GWAS) and biochemical studies have identified intracellular trafficking regulators as genetic risk factors for such pathologies. The sorting nexins (SNXs) are a family of proteins involved in protein trafficking regulation and signaling. SNXs dysregulation occurs in patients with Alzheimer’s disease (AD), Down’s syndrome (DS), schizophrenia, ataxia and epilepsy, among others, establishing clear roles for this protein family in pathology. Interestingly, restoration of SNXs levels has been shown to trigger synaptic plasticity recovery in a DS mouse model. This review encompasses an historical and evolutionary overview of SNXs protein family, focusing on its organization, phyla conservation, and evolution throughout the development of the nervous system during speciation. We will also survey SNXs molecular interactions and highlight how defects on SNXs underlie distinct pathologies of the CNS. Ultimately, we discuss possible strategies of intervention, surveying how our knowledge about the fundamental processes regulated by SNXs can be applied to the identification of novel therapeutic avenues for SNXs-related disorders.

The Red Neuronal Pigment in the Nervous System of the Mussel, Mytilus Edulis: Localization and Its Effect on Lateral Ciliary Activity with Spectral and Analytical Changes

1981 ◽  
Vol 39 ◽  
pp. 494-495
Author(s):  
Anthony A. Paparo ◽  
Judith A. Murphy
Keyword(s):  
Nervous System ◽  
Mytilus Edulis ◽  
Neuronal Cell ◽  
Ciliary Activity ◽  
Neuronal Cell Bodies ◽  
The Central Nervous System ◽  
Intracellular Organelles ◽  
The Common ◽  
The Individual ◽  
Cryostat Sections

The purpose of this study was to localize the red neuronal pigment in Mytilus edulis and examine its role in the control of lateral ciliary activity in the gill. The visceral ganglia (Vg) in the central nervous system show an over al red pigmentation. Most red pigments examined in squash preps and cryostat sec tions were localized in the neuronal cell bodies and proximal axon regions. Unstained cryostat sections showed highly localized patches of this pigment scattered throughout the cells in the form of dense granular masses about 5-7 um in diameter, with the individual granules ranging from 0.6-1.3 um in diame ter. Tissue stained with Gomori's method for Fe showed bright blue granular masses of about the same size and structure as previously seen in unstained cryostat sections.Thick section microanalysis (Fig.l) confirmed both the localization and presence of Fe in the nerve cell. These nerve cells of the Vg share with other pigmented photosensitive cells the common cytostructural feature of localization of absorbing molecules in intracellular organelles where they are tightly ordered in fine substructures.

Freeze-substitution of rye grass and ragweed pollen grains

1990 ◽  
Vol 48 (3) ◽  
pp. 686-687
Author(s):  
Liza B. Martinez ◽  
Susan M. Wick
Keyword(s):  
Cell Function ◽  
Pollen Grains ◽  
Freeze Substitution ◽  
Cell Types ◽  
Rapid Freezing ◽  
Rye Grass ◽  
Acrylic Resins ◽  
Allergenic Proteins ◽  
Cold Embedding ◽  
Structural Preservation

Rapid freezing and freeze-substitution have been employed as alternatives to chemical fixation because of the improved structural preservation obtained in various cell types. This has been attributed to biomolecular immobilization derived from the extremely rapid arrest of cell function. These methods allow the elimination of conventionally used fixatives, which may have denaturing or “masking” effects on proteins. Thus, this makes them ideal techniques for immunocytochemistry, in which preservation of both ultrastructure and antigenicity are important. These procedures are also compatible with cold embedding acrylic resins which are known to increase sensitivity in immunolabelling.This study reveals how rapid freezing and freeze-substitution may prove to be useful in the study of the mobile allergenic proteins of rye grass and ragweed. Most studies have relied on the use of osmium tetroxide to achieve the necessary ultrastructural detail in pollen whereas those that omitted it have had to contend with poor overall preservation.

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