The use of the human neuroblastoma SH-SY5Y to study the effect of second messengers on noradrenaline release

Vaughan, Peter F. T.; Peers, Chris; Walker, John H.

doi:10.1016/0306-3623(94)00312-b

Cited by 72 publications

(48 citation statements)

References 56 publications

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“…We chose to experiment with SH-SY5Y, because this neuroblastoma cell line derived from the sympathetic nervous system expresses several properties of mature sympathetic neurons (10). In addition, our findings in neuroblastoma cells were strengthened by their similarity to the findings obtained in native cardiac sympa- thetic terminals.…”

Section: Discussionmentioning

confidence: 97%

“…The exocytotic release of norepinephrine from postganglionic sympathetic neurons requires entry of Ca 2ϩ through voltagedependent Ca 2ϩ channels (10,18,19). Having first established that H 3 R activation down-regulates norepinephrine exocytosis from cardiac sympathetic terminals (1, 2, 7), we subsequently found that the selective N-type Ca 2ϩ channel blocker -CTX (8) potentiates the effects of H 3 R activation (1).…”

Section: Discussionmentioning

confidence: 99%

“…The human neuroblastoma cell line SH-SY5Y has been used as a model to study mechanisms of neurotransmitter release (10). To determine whether H 3 R activation modulates norepinephrine release by impeding Ca 2ϩ influx, we stably transfected SH-SY5Y cells with the cDNA for the H 3 R (SH-SY5Y-H 3 ).…”

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confidence: 99%

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Decreased intracellular calcium mediates the histamine H ₃ -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings

Silver

Poonwasi

Seyedi

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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Activation of presynatic histamine H3 receptors (H3R) down-regulates norepinephrine exocytosis from cardiac sympathetic nerve terminals, in both normal and ischemic conditions. Analogous to the effects of ␣2-adrenoceptors, which also act prejunctionally to inhibit norepinephrine release, H 3R-mediated antiexocytotic effects could result from a decreased Ca 2؉ influx into nerve endings. We tested this hypothesis in sympathetic nerve terminals isolated from guinea pig heart (cardiac synaptosomes) and in a model human neuronal cell line (SH-SY5Y), which we stably transfected with human H 3R cDNA (SH-SY5Y-H3). We found that reducing Ca 2؉ influx in response to membrane depolarization by inhibiting Ntype Ca 2؉ channels with -conotoxin ( -CTX) greatly attenuated the exocytosis of W e have found that sympathetic nerve endings in the guinea pig (1) and human heart (2) express the H 3 histamine receptor subtype (H 3 R). Activation of these H 3 R down-regulates norepinephrine exocytosis in both normal and ischemic conditions (3, 4).The H 3 R is a G-protein-coupled receptor linked to the inhibition of adenylyl cyclase (5). The H 3 R is modestly related in sequence to most biogenic amine receptors, including ␣ 2 -adrenoceptors (6). Like the H 3 R, cardiac ␣ 2 -adrenoceptors act prejunctionally to inhibit norepinephrine exocytosis (3, 7). Moreover, inhibition of Ca 2ϩ influx into sympathetic nerve endings with the selective N-type Ca 2ϩ channel blocker -conotoxin ( -CTX) (8) potentiates both H 3 R-and ␣ 2 -adrenoreceptor-mediated attenuation of cardiac adrenergic responses (1). Therefore, it is conceivable that, as for ␣ 2 -adrenoceptors (9), H 3 R activation may also inhibit norepinephrine release by diminishing Ca 2ϩ influx into cardiac sympathetic terminals.The human neuroblastoma cell line SH-SY5Y has been used as a model to study mechanisms of neurotransmitter release (10). To determine whether H 3 R activation modulates norepinephrine release by impeding Ca 2ϩ influx, we stably transfected SH-SY5Y cells with the cDNA for the H 3 R (SH-SY5Y-H 3 ). We report that upon K ϩ -induced depolarization, activation of H 3 R is associated with a marked decrease in intracellular Ca 2ϩ (Ca i ) and, thus, with an attenuation of norepinephrine exocytosis. Materials and MethodsPreparation of Cardiac Synaptosomes. Male Hartley guinea pigs (Charles River Breeding Laboratories) weighing 250-300 g were killed by cervical dislocation under light anesthesia with CO 2 vapor. The rib cage was rapidly opened and the heart was dissected away. A cannula was inserted into the aorta, and the heart was perfused for 5 min at constant pressure (40 cm H 2 O) in a Langendorff apparatus (11) with Ringer's solution (154 mM NaCl͞5.61 mM KCl͞2.16 mM CaCl 2 ͞5.95 mM NaHCO 3 ͞5.55 mM glucose) equilibrated with 100% O 2 at 37°C. This procedure ensured that no blood traces remained in the coronary circulation. Hearts were then freed from fat and connective tissue and minced in ice-cold 0.32 M sucrose containing 1 mM EGTA (pH 7.4). Synaptosomes were isolated as...

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Decreased intracellular calcium mediates the histamine H ₃ -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings

Silver

Poonwasi

Seyedi

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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“…3D). This was unexpected because the predominant muscarinic receptor type in SH-SY5Y cells is m3 (32,33), which via phosphoinositidase C generates InsP 3 and diacylglycerol and thus elevates [Ca 2ϩ ] i and protein kinase C activity (33). The effect of carbachol (IC 50 ϭ ϳ1 M) was blocked by atropine and was not additive with a maximal concentration of VIP, suggesting that VIP and carbachol act via the same pathway (Fig.…”

Section: Phosphorylation Of Immunoprecipitated Type I Ii and Iii Inspmentioning

confidence: 99%

Phosphorylation of Inositol 1,4,5-Trisphosphate Receptors by cAMP-dependent Protein Kinase

Wojcikiewicz

Luo

1998

Journal of Biological Chemistry

147

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The ability of cAMP-dependent protein kinase (PKA) to phosphorylate type I, II, and III inositol 1,4,5-trisphosphate (InsP 3 ) receptors was examined. The receptors either were immunopurified from cell lines and then phosphorylated with purified PKA or were phosphorylated in intact cells after activating intracellular cAMP formation. The former studies showed that the type I receptor was a good substrate for PKA (0.65 mol P i incorporated/mol receptor), whereas type II and III receptors were phosphorylated relatively weakly. The latter studies showed that despite these differences, each of the receptors was phosphorylated in intact cells in response to forskolin or activation of neurohormone receptors. Detailed examination of SH-SY5Y neuroblastoma cells, which express >99% type I receptor, revealed that minor increases in cAMP concentration were sufficient to cause maximal phosphorylation. Thus, VIP and pituitary adenylyl cyclase activating peptide (acting through G s -coupled pituitary adenylyl cyclase activating peptide-I receptors) were potent stimuli of type I receptor phosphorylation, and remarkably, even slight increases in cAMP concentration induced by carbachol (acting through G q -coupled muscarinic receptors) or other Ca 2؉ mobilizing agents were sufficient to cause phosphorylation. Finally, PKA enhanced InsP 3 -induced Ca 2؉ mobilization in a range of permeabilized cell types, irrespective of whether the type I, II, or III receptor was predominant. In summary, these data show that all InsP 3 receptors are phosphorylated by PKA, albeit with marked differences in stoichiometry. The ability of both G s -and G q -coupled cell surface receptors to effect InsP 3 receptor phosphorylation by PKA suggests that this process is widespread in mammalian cells and provides multiple routes by which the cAMP signaling pathway can influence Ca 2؉ mobilization.Inositol 1,4,5-trisphosphate (InsP 3 ) 1 receptors form tetrameric channels in endoplasmic reticulum membranes that conduct Ca 2ϩ in an InsP 3 -sensitive manner (1-3). Thus, they link cell surface receptor-mediated increases in InsP 3 formation to increases in cytoplasmic free Ca 2ϩ concentration ([Ca 2ϩ ] i ). To date, the coding regions of three mammalian InsP 3 receptor genes have been sequenced (4 -11). Their products, termed type I, II, and III receptors, are ϳ2700 amino acids in length and are 60 -70% identical at the amino acid level (2-11). Each receptor is thought to have the same overall structure, being divided into three domains: an N-terminal ligand binding domain, a C-terminal channel forming domain, and an intervening sequence, termed the coupling domain, that contains sites either known or hypothesized to be involved in receptor regulation (2-4, 6, 12). Type I, II, and III receptors are expressed in different amounts in different cell types, and some cells co-express all three receptors (13,14). InsP 3 receptors form heterotetramers in intact cells, and such associations persist after receptor solubilization (14 -17).Characterization of diffe...

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“…SH-SY5Y cells are derived from human sympathetic neuronal tissue, and maintain many properties of nerve cells, thus providing a useful model for the characterization of molecules affecting human neuronal function, including endogenously expressed calcium channels [15][16][17][18][19]. In order to deplete TAF1 expression in vitro, TAF1-specific siRNAs were transfected into SH-SY5Y cells, and depletion validated using qRT-PCR.…”

Section: Neuronal Ion Channel Gene Expression Analysismentioning

confidence: 99%

A novel variant in TAF1 affects gene expression and is associated with X-linked TAF1 intellectual disability syndrome

et al. 2018

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We investigated the genome of a 5-year old male who presented with global developmental delay (motor, cognitive, and speech), hypotonia, possibly ataxia, and cerebellar hypoplasia of unknown origin. Whole genome sequencing and mRNA-sequencing were performed on a family containing an affected proband, his unaffected parents and maternal grandfather. To explore the molecular and functional consequences of the variant, we performed cell proliferation assays, qRT-PCR array, immunoblotting, calcium imaging, and neurite outgrowth experiments in SH-SY5Y neuroblastoma cells to compare the properties of the wild type TAF1, deletion of TAF1, and TAF1 variant p.Ser1600Gly samples. The whole genome data identified several gene variants. However, the genome sequence data failed to implicate a candidate gene as many of the variants were of unknown significance. By combining genome sequence data with transcriptomic data, a probable candidate variant, p.Ser1600Gly, emerged in TAF1. Moreover, the RNA-seq data revealed a 90:10 extremely skewed X-chromosome inactivation in the mother. Our results show that neuronal ion channel genes were differentially expressed between TAF1 deletion and TAF1 variant p.Ser1600Gly cells, when compared to their respective controls, and that the TAF1 variant may impair neuronal differentiation and cell proliferation. Taken together, our data suggests that this novel variant in TAF1 plays a key role in the development of a recently described X-linked syndrome, TAF1 intellectual disability syndrome, and further extends our knowledge of a potential link between TAF1 deficiency and defects in neuronal cell function.

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The use of the human neuroblastoma SH-SY5Y to study the effect of second messengers on noradrenaline release

Cited by 72 publications

References 56 publications

Decreased intracellular calcium mediates the histamine H ₃ -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings

Decreased intracellular calcium mediates the histamine H ₃ -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings

Phosphorylation of Inositol 1,4,5-Trisphosphate Receptors by cAMP-dependent Protein Kinase

A novel variant in TAF1 affects gene expression and is associated with X-linked TAF1 intellectual disability syndrome

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The use of the human neuroblastoma SH-SY5Y to study the effect of second messengers on noradrenaline release

Cited by 72 publications

References 56 publications

Decreased intracellular calcium mediates the histamine H 3 -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings

Decreased intracellular calcium mediates the histamine H 3 -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings

Phosphorylation of Inositol 1,4,5-Trisphosphate Receptors by cAMP-dependent Protein Kinase

A novel variant in TAF1 affects gene expression and is associated with X-linked TAF1 intellectual disability syndrome

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Decreased intracellular calcium mediates the histamine H ₃ -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings

Decreased intracellular calcium mediates the histamine H ₃ -receptor-induced attenuation of norepinephrine exocytosis from cardiac sympathetic nerve endings