The dispersed cell culture as model for functional studies of the subcommissural organ: preparation and characterization of the culture system

Schöniger, Sandra; Wehming, Sascha; González, César; Schöbitz, Karin; Rodríguez, Estéban M.; Oksche, A.; Yulis, Carlos R.; Nürnberger, F.

doi:10.1016/s0165-0270(01)00351-x

Cited by 22 publications

(26 citation statements)

References 46 publications

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“…It will be of interest to determine whether any of these previously described mutants have abnormalities in the expression or sequence of the RFX4_v3 protein. In addition, examination of downstream gene expression at this specific anatomical site in these hemizygous mice may lead to new insights into the formation of this 'enigmatic secretory gland of the brain' (Schoniger et al, 2001). …”

Section: Discussionmentioning

confidence: 99%

Graded phenotypic response to partial and complete deficiency of a brain-specific transcript variant of the winged helix transcription factor RFX4

et al. 2003

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Interruption of a single allele prevented formation of the subcommissural organ, a structure important for cerebrospinal fluid flow through the aqueduct of Sylvius, and resulted in congenital hydrocephalus. These data implicate the RFX4_v3 variant transcript as being crucial for early brain development, as well as for the genesis of the subcommissural organ. These findings may be relevant to human congenital hydrocephalus, a birth defect that affects ~0.6 per 1000 newborns.

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Section: Discussionmentioning

confidence: 99%

Graded phenotypic response to partial and complete deficiency of a brain-specific transcript variant of the winged helix transcription factor RFX4

et al. 2003

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“…Cultures were maintained at 37°C in a humidified atmosphere with 5% CO 2 . The mitotic inhibitor cytosine β-arabino furanoside (1 μM) (Sigma-Aldrich, St. Louis, MO) was added to the culture to inhibit glial cell proliferation (Schoniger et al, 2001). Cultures were maintained by changing the media every 2-3 days.…”

Section: Cerebrocortical Neuronal Culturementioning

confidence: 99%

Excitatory effects of human immunodeficiency virus 1 Tat on cultured rat cerebral cortical neurons

et al. 2008

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HIV-1 Tat protein is one of the neurotoxins involved in the pathogenesis of HIV-1-associated neuronal disorders. Combined electrophysiological and optical imaging experiments were undertaken to investigate whether HIV-1 Tat30-86, herein referred to as Tat30-86, acted directly or indirectly via the release of glutamate or both and to test its effect on the properties of spontaneous quantal events in cultured cortical neurons. Whole-cell patch recordings were made from cultured rat cortical neurons in either current-or voltage-clamp mode. Tat30-86 (50-1000 nM) induced in a population of cortical neurons a long-lasting depolarization, which was accompanied by a decrease of membrane resistance and persisted in a Krebs solution containing tetrodotoxin (TTX, 0.5 μM). Depolarizations were slightly reduced by pretreatment with glutamate receptor antagonists CNQX (10 μM) and AP-5 (50 μM), and were markedly reduced in a Ca 2+ -free Krebs solution; the differences were statistically significant. Tat30-86-induced inward currents had a reversal potential between -30 and 0 mV. While not causing a noticeable depolarization, lower concentrations of Tat30-86 (10 nM) increased membrane excitability, as indicated by increased numbers of neuronal discharge in response to a step depolarizing pulse. Tat30-86 (10 nM) increased the frequency of spontaneous miniature excitatory postsynaptic currents (mEPSCs), while not significantly affecting their amplitude. Tat30-86 (10 nM) moderately increased the frequency as well as the amplitude of spontaneous miniature inhibitory postsynaptic currents (mIPSCs). Ratiometric Ca 2+ imaging studies showed that Tat30-86 produced three types of Ca 2+ responses: 1) a fast and transitory increase, 2) Ca 2+ oscillations, and 3) a fast increase followed by a plateau; the glutamate receptor antagonists eliminated the late component of Ca 2+ response. The result suggests that Tat30-86 is an active fragment and that it excites cortical neurons directly and indirectly via releasing glutamate from adjacent neurons. Keywords calcium imaging; electrophysiology; postsynaptic currentsInfection of the central nervous system (CNS) with the human immunodeficiency virus 1 (HIV-1) may lead to cognitive, motor and behavioral dysfunctions, collectively termed HIVCorresponding author: G. Cristina Brailoiu, M.D., Department of Pharmacology, Temple University School of Medicine, 3420 N. Broad Street, Philadelphia, PA 19140 USA, Tel: 215-707-7705, Fax: 215-707-7068, E-mail: gbrailou@temple.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. (Kaul et al., 2001; Gonzales-Scara...

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“…Furthermore, we have studied the effects on CREB phosphorylation elicited by serotonin (5HT) and substance P (SP), which have previously been shown to be present in nerve fibers innervating the SCO (Ljungdahl et al 1978;Møllgård and Wiklund 1979;Léger et al 1983;Bouchaud 1993;Knigge and Schock 1993;Mikkelsen et al 1997;Schöniger et al 2001) or by ATP, an important neurotransmitter in the CNS and peripheral nervous system, shown to activate neurons and glial cells (for a review, see Zimmermann 1994) including SCO cells (Schöniger et al 2002).…”

Section: Introductionmentioning

confidence: 99%

“…The experiments have been performed with explant and dissociated cell cultures of the SCO from the bovine, because the SCO secretory glycoproteins of this species have been extensively characterized in previous studies (Rodríguez et al 1987;Meiniel et al 1988;Nualart et al 1991;Hein et al 1993), and the bovine SCO can be readily isolated and cultured Schöniger et al 2001Schöniger et al , 2002.…”

Section: Introductionmentioning

confidence: 99%

Transcription factor CREB and its stimulus-dependent phosphorylation in cell and explant cultures of the bovine subcommissural organ

Schöniger

Maronde

Kopp

et al. 2002

Cell and Tissue Research

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The subcommissural organ (SCO) is an ependymal brain gland that synthesizes and secretes glycoproteins. Very little is known about the signal transduction cascades operating in this organ and their impact on gene expression. An important transcription factor that regulates gene expression in glial cells and neurons is the cyclic-AMP-responsive element binding protein (CREB), which is activated by phosphorylation of the serine residue 133. Here, we analyzed the presence of CREB in bovine SCO cells and its phosphorylation by drugs that activate cyclic-AMP-dependent or calcium-dependent signal transduction pathways. We also investigated the effects of three natural signaling molecules, serotonin (5HT), substance P (SP) and ATP, on CREB phosphorylation and on the second messengers cyclic AMP and calcium. Investigations were performed with cell and explant cultures by using immunocytochemistry, immunoblot, enzyme-linked immunosorbent assay, and the Fura-2 technique. A strong immunosignal for total (phosphorylated and unphosphorylated) CREB was found in virtually all SCO cells. Total CREB levels did not change upon stimulation. Phosphorylated (p)CREB levels were low in unstimulated cells and significantly elevated by drugs that increase the levels of cyclic AMP or free calcium ions. pCREB was also induced by SP and ATP; both substances increased the intracellular calcium concentration but did not affect the formation of intracellular cyclic AMP. 5HT did not influence the phosphorylation of CREB, the intracellular calcium concentration, or the formation of cyclic AMP. Our data identify CREB as an SCO transcription factor that can be activated by the second messengers cAMP and calcium. SP and ATP stimulate the phosphorylation of CREB apparently via a calcium-dependent mechanism and are thus involved in the control of gene expression in the bovine SCO.

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The dispersed cell culture as model for functional studies of the subcommissural organ: preparation and characterization of the culture system

Cited by 22 publications

References 46 publications

Graded phenotypic response to partial and complete deficiency of a brain-specific transcript variant of the winged helix transcription factor RFX4

Graded phenotypic response to partial and complete deficiency of a brain-specific transcript variant of the winged helix transcription factor RFX4

Excitatory effects of human immunodeficiency virus 1 Tat on cultured rat cerebral cortical neurons

Transcription factor CREB and its stimulus-dependent phosphorylation in cell and explant cultures of the bovine subcommissural organ

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