Pharmacological Characterization of Cholinergic Receptors in a Human Neuroblastoma Cell Line

Clementi, Francesco; Cabrini, D.; Gotti, Cecilia; Sher, Emanuele

doi:10.1111/j.1471-4159.1986.tb02861.x

Cited by 55 publications

(6 citation statements)

References 23 publications

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“…The neuroblastoma cell line IMR-32 is an adrenergic human cell line that can be differentiated in vitro to extend long axon-like processes with numerous growth cones (Clementi et al, 1986;Carbone et al, 1990). After ϳ1 week of treatment of IMR-32 cell cultures with BrdU, the cell proliferation had ceased and an extensive network of processes was visible (data not shown).…”

Section: Baculovirally Expressed Ox 1 Receptors In Differentiated Imrmentioning

confidence: 99%

The Orexin OX₁Receptor Regulates Ca²⁺Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells

Näsman¹,

Bart²,

Larsson³

et al. 2006

J. Neurosci.

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Section: Baculovirally Expressed Ox 1 Receptors In Differentiated Imrmentioning

confidence: 99%

The Orexin OX₁Receptor Regulates Ca²⁺Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells

Näsman¹,

Bart²,

Larsson³

et al. 2006

J. Neurosci.

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“…Sequence from four primary NB tumors (Lastowska et al 2007), four ALL samples (Case et al 2008), and the NB cell line IMR32 (Clementi et al 1986) was generated, originally to develop and assess mutation detection pipelines. Appropriate informed consent for use of all primary material was obtained (Lastowska et al 2007, Case et al 2008.…”

Section: Transcriptome Sequencingmentioning

confidence: 99%

Post-transcriptional exon shuffling events in humans can be evolutionarily conserved and abundant

Al-Balool¹,

Weber²,

Liu³

et al. 2011

Genome Res.

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In silico analyses have established that transcripts from some genes can be processed into RNAs with rearranged exon order relative to genomic structure (post-transcriptional exon shuffling, or PTES). Although known to contribute to transcriptome diversity in some species, to date the structure, distribution, abundance, and functional significance of human PTES transcripts remains largely unknown. Here, using high-throughput transcriptome sequencing, we identify 205 putative human PTES products from 176 genes. We validate 72 out of 112 products analyzed using RT-PCR, and identify additional PTES products structurally related to 61% of validated targets. Sequencing of these additional products reveals GT-AG dinucleotides at >95% of the splice junctions, confirming that they are processed by the spliceosome. We show that most PTES transcripts are expressed in a wide variety of human tissues, that they can be polyadenylated, and that some are conserved in mouse. We also show that they can extend into 59 and 39 UTRs, consistent with formation via trans-splicing of independent pre-mRNA molecules. Finally, we use real-time PCR to compare the abundance of PTES exon junctions relative to canonical exon junctions within the transcripts from seven genes. PTES exon junctions are present at <0.01% to >90% of the levels of canonical junctions, with transcripts from MAN1A2, PHC3, TLE4, and CDK13 exhibiting the highest levels. This is the first systematic experimental analysis of PTES in human, and it suggests both that the phenomenon is much more widespread than previously thought and that some PTES transcripts could be functional.

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“…Human neuroblastoma IMR-32 cells respond to treatment with BrdU by acquiring a neuronal phenotype exhibiting a wide variety of voltage gated calcium channels (N-, and T-type) as well as the outgrowth of long axon-like extensions (Clementi et al, 1986;Carbone et al, 1990). We tested for the presence TRPA1 channels mRNA in non-differentiated and differentiated IMR-32 cells.…”

Section: Expression Of Trpa1 Channels In Differentiated Imr-32 Cellsmentioning

confidence: 99%

“…The most notable changes are the selective expression of voltage gated calcium channels almost exclusively of N-type type (Clementi et al, 1986;Carbone et al, 1990). As discussed above TRPA1 and TRPM8 channel expression is developmentally regulated.…”

mentioning

confidence: 98%

Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR‐32 human neuroblastoma cells

Louhivuori

Bart

Larsson

et al. 2009

Journal Cellular Physiology

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TRPA1 and TRPM8 are transient receptor potential (TRP) channels involved in sensory perception. TRPA1 is a non-selective calcium permeable channel activated by irritants and proalgesic agents. TRPM8 reacts to chemical cooling agents such as menthol. The human neuroblastoma cell line IMR-32 undergoes a remarkable differentiation in response to treatment with 5-bromo-2-deoxyuridine. The cells acquire a neuronal morphology with increased expression of N-type voltage gated calcium channels and neurotransmitters. Here we show using RT-PCR, that mRNA for TRPA1 and TRPM8 are strongly upregulated in differentiating IMR-32 cells. Using whole cell patch clamp recordings, we demonstrate that activators of these channels, wasabi, allyl-isothiocyanate (AITC) and menthol activate membrane currents in differentiated cells. Calcium imaging experiments demonstrated that AITC mediated elevation of intracellular calcium levels were attenuated by ruthenium red, spermine, and HC-030031 as well as by siRNA directed against the channel. This indicates that the detected mRNA level correlate with the presence of functional channels of both types in the membrane of differentiated cells. Although the differentiated IMR-32 cells responded to cooling many of the cells showing this response did not respond to TRPA1/TRPM8 channel activators (60% and 90% for AITC and menthol respectively). Conversely many of the cells responding to these activators did not respond to cooling (30%). This suggests that these channels have also other functions than cold perception in these cells. Furthermore, our results suggest that IMR-32 cells have sensory characteristics and can be used to study native TRPA1 and TRPM8 channel function as well as developmental expression.

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Pharmacological Characterization of Cholinergic Receptors in a Human Neuroblastoma Cell Line

Cited by 55 publications

References 23 publications

The Orexin OX₁Receptor Regulates Ca²⁺Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells

The Orexin OX₁Receptor Regulates Ca²⁺Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells

Post-transcriptional exon shuffling events in humans can be evolutionarily conserved and abundant

Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR‐32 human neuroblastoma cells

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Pharmacological Characterization of Cholinergic Receptors in a Human Neuroblastoma Cell Line

Cited by 55 publications

References 23 publications

The Orexin OX1Receptor Regulates Ca2+Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells

The Orexin OX1Receptor Regulates Ca2+Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells

Post-transcriptional exon shuffling events in humans can be evolutionarily conserved and abundant

Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR‐32 human neuroblastoma cells

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The Orexin OX₁Receptor Regulates Ca²⁺Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells

The Orexin OX₁Receptor Regulates Ca²⁺Entry via Diacylglycerol-Activated Channels in Differentiated Neuroblastoma Cells