Cholinergic receptors, ion channels, neurotransmitter synthesis, and neurite outgrowth are independently regulated during the in vitro differentiation of a human neuroblastoma cell line

Gotti, Cecilia; Sher, Emanuele; Cabrini, D.; Bondiolotti, Gianpietro; Wanke, Enzo; Mancinelli, E.; Clementi, Francesco

doi:10.1111/j.1432-0436.1987.tb00061.x

Cited by 61 publications

(44 citation statements)

References 27 publications

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“…Differentiation characteristics include morphological, biochemical and electrophysiological parameters. Neurally differentiated cells show a typically neural-like appearance [12,34,41], express neuron-specific markers, while glial-specific markers are absent (present study) and behave as neurons in electrophysiologic studies [1,12]. The production of neuropeptides and the expression of chromogranin B mRNA [10] can be viewed as another characteristic of neuroendocrine differentiation.…”

Section: Discussionmentioning

confidence: 66%

“…The presence of NSE mRNA and the absence of GFAP, NF and PLP mRNAs in IMR32 cells irrespective of differentiation suggests that these cells are committed to a neural cell pathway. Neuronally-differentiated neuroblastoma cells have been shown to contain neurotransmitter receptors [12,24] and enzymes [12,13,41]. They produce a number of neurotransmitters such as dopamine, noradrenaline and serotonin [13], and contain noradrenergic uptake systems [30].…”

Section: Discussionmentioning

confidence: 99%

“…hypothalamic neurons, to establish cell culture systems producing hypothalamic hormones. Indeed, we recently reported that the differentiating agent bromo-deoxy-uridine that is known to induce differentiation in neuroblastoma cells [12], markedly enhances production of TRH in fetal hypothalamic neurones in primary culture [6].…”

Section: Discussionmentioning

confidence: 99%

“…These ceils have been reported to produce different neuropeptides [5,26,27,36,37] and neurotransmitters [12,13,34] and thus constitute an alternative model for neural peptide synthesis.…”

Section: Introductionmentioning

confidence: 99%

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Neural differentiation of the human neuroblastoma cell line IMR32 induces production of a thyrotropin-releasing hormone-like peptide

Rondeel¹,

Klootwijk

Linkels

et al. 1994

Brain Research

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The human neuroblastoma cell line IMR32 produces and secretes substantial amounts of TRH-immunoreactivity (TRH-IR) as measured with radioimmunoassay (RIA) using the nonspecific antiserum 4319. It was found that synthesis of TRH-IR is dependent on neural differentiation: under serum-free conditions these cells exhibit neural characteristics as defined by morphological and biochemical standards. After culture for 2-5 days in serum-free medium cells grew large neural processes and expressed neuron-specific markers whereas glial-specific markers were absent. TRH-IR became detectable after 4-8 days serum-free conditions. Northern blot and chromatographic analysis, however, failed to detect proTRH mRNA and authentic TRH in these cells. Moreover, TRH-IR was undetectable in the RIA using TRH-specific antiserum 8880. TRH-IR produced by differentiated cells was retained on a QAE Sephadex A-25 anion-exchange column and thus negatively charged. HPLC analysis showed coelution with the synthetic peptide pGlu-Glu-ProNH e. Study of the mechanisms regulating production of this novel peptide in these cells should further elucidate the role differentiation plays in the synthesis of neuropeptides.

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Section: Discussionmentioning

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…These ceils have been reported to produce different neuropeptides [5,26,27,36,37] and neurotransmitters [12,13,34] and thus constitute an alternative model for neural peptide synthesis.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Neural differentiation of the human neuroblastoma cell line IMR32 induces production of a thyrotropin-releasing hormone-like peptide

Rondeel¹,

Klootwijk

Linkels

et al. 1994

Brain Research

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“…As reported by Gotti et al (1987), when the cell was under voltage clamp and the pipette contained a high concentration of potassium (solution G, Methods), the response to depolarization from a sufficiently negative holding potential consisted of a rapid inward followed by a sustained outward current. The sustained outward component was not seen if the potassium in the pipette was replaced with caesium (solution H, Methods); it could therefore be identified as a potassium current.…”

Section: Voltage Clamp Sodium Currentsmentioning

confidence: 78%

On the sodium and potassium currents of a human neuroblastoma cell line.

Ginsborg

Martin

Patmore

1991

The Journal of Physiology

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SUMMARY1. The patch-clamp method was applied to the study of ionic currents activated by depolarization of undifferentiated IMR-32 human neuroblastoma cells. Wholecell sodium and potassium currents and single potassium ion channel currents from cell-attached patches were investigated.2. Cells had a mean resting potential of -38 mV and mean input resistance of 16 GQ. Single action potentials were evoked under current clamp during the injection of depolarizing currents.3. A voltage-dependent inward sodium current was observed which reversed at +44 mV. A Boltzmann fit to the activation curve gave a half-maximal activation voltage of -41-6 mV and a 'slope' of 3.9 mV. The steady-state inactivation curve had a half-maximal inactivation voltage of -81 mV and a 'slope' of 9-7 mV.4. The time-dependent activation and inactivation of the current displayed cl'assical Hodgkin-Huxley kinetics. Values for the time constants Tm and Th of 0-16 and 0-63 ms were calculated for a voltage jump from -80 to -10 mV; Tm and Th decreased as the step potential was changed from -30 to + 20 mV.5. Outward currents were activated in bathing solutions substantially free of anions and could thus be attributed to potassium ions. The tail current reversed in direction on repolarization to -60 mV when the potassium concentration in the bathing solution was increased from 6 to 30 mm. When the bathing solution contained 145 mM-potassium, and the patch pipette, 95 mm, a depolarization to -10 mV from a holding potential of -60 mV evoked an inward current.6. Outward currents were examined by using voltage pulses which depolarized the cell to -20 mV, or more positive values, from a holding potential of -80 mV and by pulses which depolarized the cell to 0 mV, or to positive values, from a holding potential of -30 mV. A Boltzmann fit of typical activation data gave a halfmaximal activation voltage of 17 mV and a 'slope' of 14 mV.7. The time course of the rising phase of the current was described by a function of the form A{1 -exp -(t-6t)/r]}, where 6t varied between 1 and 4 ms and r varied between 4 and 27 ms, decreasing with increasing depolarization. There was no evidence for a fast transient component.M,S1 8428 B. L. GINSBORG, R. J. MARTIN AND L. PATMORE 8. The amplitude of outward currents was reduced by extracellular calcium ions, cobalt ions, tetraethylammonium and 4-aminopyridine.9. During prolonged depolarization, the time course of the current could be described by the sum of two exponentially decaying components, with time constants of around 1 and 10 s. 4-Aminopyridine selectively reduced the slower component.10. Single-channel currents were evoked by depolarization of cell-attached patches with electrodes containing normal extracellular solution. The most frequently encountered channel had a conductance of about 20 pS. Patches usually contained either no or several such channels; the activity of a single channel, however, could be recorded during the inactivation which occurred during prolonged depolarization.11. A detailed study was made of r...

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Mitogenic effect of neurotrophic factors on human imr 32 neuroblastoma cells

Lüdecke

Unsicker

1990

Cancer

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The effect of growth factors with neurotrophic properties on proliferation of the human IMR 32 neuroblastoma (NB) cell line was studied. A colorimetric proliferation assay and an anchorage-dependent cell culture system were used. Basic fibroblast growth factor (bFGF), nerve growth factor (NGF), neurite-inducing factor (NIF), ciliary neurotrophic factor (CNTF), and a cell-free extract from selected embryonic chick eye tissues (CIPE) were assayed for their capacity to control proliferation. Basic FGF, NGF, and CIPE stimulated proliferation of IMR 32 NB cells in serum-containing culture conditions. The NIF and CNTF had no effect. The concentration of bFGF required to induce half-maximal cell growth was 4.6 +/- 1.8 ng/ml, but the half-maximally effective dose of NGF was 7.5 +/- 2.7 ng/ml. In combination these two growth factors were additive within a small concentration range. In serum-free culture conditions bFGF affected both proliferation and cell differentiation by promoting neurite growth and cell aggregate formation. In contrast, NGF induced cell neurite outgrowth only. These results, in conjunction with the evidence that bFGF-like molecules are present, in IMR 32 NB cells may support the notion that NB cells regulate their proliferation by an autocrine mechanism. Basic FGF and NGF, two distinct neurotrophic factors, appear to be involved in the regulation of NB cell proliferation.

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Cholinergic receptors, ion channels, neurotransmitter synthesis, and neurite outgrowth are independently regulated during the in vitro differentiation of a human neuroblastoma cell line

Cited by 61 publications

References 27 publications

Neural differentiation of the human neuroblastoma cell line IMR32 induces production of a thyrotropin-releasing hormone-like peptide

Neural differentiation of the human neuroblastoma cell line IMR32 induces production of a thyrotropin-releasing hormone-like peptide

On the sodium and potassium currents of a human neuroblastoma cell line.

Mitogenic effect of neurotrophic factors on human imr 32 neuroblastoma cells

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