Tissue culture of bovine subcommissural organ

Lehmann, W.; Naumann, W.; Wagner, Ulrich

doi:10.1007/bf00174426

Cited by 13 publications

(9 citation statements)

References 38 publications

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“…The present study aims at identification of neuroactive molecules that affect intracellular signal transduction cascades in SCO cells and that may be involved in the control of the SCO secretory activity. Our investigations were performed with the bovine SCO because it can be easily dissected from the adjacent brain tissue and cultured for several weeks (Lehmann et al 1993;Schöniger et al 2001). Moreover, with regard to the secretory products of the SCO, bovine is the best-investigated vertebrate species (Rodríguez et al 1987;Estivill-Torrús et al 1998;Meiniel et al 1988;Nualart et al 1991;Hein et al 1993).…”

Section: Discussionmentioning

confidence: 99%

“…To analyze calcium signal transduction cascades, isolated SCO cells were prepared either from whole SCOs freshly dissected from the adjacent posterior commissure and cut into small pieces (~1 mm in diameter) or from SCO explant cultures that had initially been cultured for 1-2 months according to the method of Lehmann et al (1993) and Schöniger et al (2001). The tissue was dissociated by papain digestion as described (Schaad et al 1993;Schomerus et al 1995;Kopp et al 1999).…”

Section: Sco Cell Culturementioning

confidence: 99%

“…Coverslips were finally mounted onto glass slides using Kaiser's Glyceringelatine. Immunocytochemical and functional data were correlated at the level of single cells; the location of the cells was determined by means of the grid etched onto the coverslip (Schomerus et al 1995;Kroeber et al 1997;Kopp et al 1999 Lehmann et al (1993) and Schöniger et al (2001). The explants were cultured in an incubator at 37°C in an atmosphere of 7% CO 2 and 93% air using supplemented DMEM/F12 as the culturing medium.…”

Section: Immunocytochemical Characterization Of the Analyzed Cellsmentioning

confidence: 99%

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Effects of neuroactive substances on the activity of subcommissural organ cells in dispersed cell and explant cultures

Schöniger

Kopp

Schomerus

et al. 2002

Cell and Tissue Research

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The subcommissural organ (SCO), an ependymal (glial) circumventricular organ, releases glycoproteins into the cerebrospinal fluid; however, the regulation of its secretory activity is largely unknown. To identify neuroactive substances that may regulate SCO activity, we investigated immunocytochemically identified bovine SCO cells by means of calcium imaging. This analysis was focused on: (1) serotonin (5HT) and substance P (SP), immunocytochemically shown to be present in axons innervating the bovine SCO; and (2) ATP, known to activate glial cells. 5HT had no effect on the intracellular calcium concentration ([Ca(2+)](i)), and its precise role remains to be clarified. SP elicited rises in [Ca(2+)](i) in approx. 30% and ATP in even 85% of the analyzed SCO cells. These effects were dose-dependent, involved NK(3) and P2Y(2) receptors linked to G protein and phospholipase C (PLC) activation, and could not be mimicked by forskolin or 8-bromo-cAMP. In 50% of the SP-sensitive cells, the increases in [Ca(2+)](i) comprised calcium release from thapsigargin-sensitive intracellular stores and an influx of extracellular calcium via protein kinase C (PKC)-induced opening of L-type voltage-gated calcium channels (VGCCs). In the remaining SP-sensitive cells, the increase in [Ca(2+)](i) was caused exclusively by influx of extracellular calcium via VGCCs of the L-type. In all ATP-sensitive cells the increase in [Ca(2+)](i) involved calcium release from thapsigargin-sensitive intracellular stores and a PKC-mediated influx of extracellular calcium via L-type VGCCs. Our data suggest that SP and ATP are involved in regulation of the activity of SCO cells.

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Section: Discussionmentioning

confidence: 99%

Section: Sco Cell Culturementioning

confidence: 99%

Section: Immunocytochemical Characterization Of the Analyzed Cellsmentioning

confidence: 99%

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Effects of neuroactive substances on the activity of subcommissural organ cells in dispersed cell and explant cultures

Schöniger

Kopp

Schomerus

et al. 2002

Cell and Tissue Research

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“…Explant culture of bovine SCOs SCO explant cultures were prepared according to the original protocol of Lehmann et al (1993;cf. Schöniger et al 2001).…”

Section: Methodsmentioning

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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“…SCO ependymal cells are specialized in the synthesis and secretion of highmolecular-mass glycoproteins that are released into the cerebrospinal fluid (CSF), where they aggregate to form a threadlike supramolecular structure known as Reissner's fibre (RF; Oksche 1969;Rodriguez et al 1992Rodriguez et al , 1998Meiniel 2001;Nualart and Hein 2001). Part of the SCO secretion may remain soluble in the CSF, under both physiological and experimental conditions (Irigoin et al 1990;Lehmann et al 1993;Rodriguez et al 1993;HoyoBecerra et al 2006;Vio et al 2008). SCO ependymal cells display long basal processes ending on blood vessels or on the leptomeningeal surface.…”

Section: Introductionmentioning

confidence: 96%

The subcommissural organ and the development of the posterior commissure in chick embryos

et al. 2009

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The subcommissural organ (SCO) is an ependymal differentiation located in the diencephalon under the posterior commissure (PC). SCO-spondin, a glycoprotein released by the SCO, belongs to the thrombospondin superfamily and shares molecular domains with axonal pathfinding molecules. Several lines of evidence suggest a relationship between the SCO and the development of the PC in the chick: (1) their close location to each other, (2) their differentiation at the same developmental stage in the chick, (3) the abnormal PC found in null mutants lacking an SCO and (4) the release by the SCO of SCO-spondin. By application of DiI crystals in the PC of chick embryos, we have identified the neurons that give rise to the PC. Labelling is confined to the magnocellular nucleus of the PC (MNPC). To gain insight into the role of the SCO in PC development, coculture experiments of explants of the MNPC region (MNPCr) from embryos at embryonic day 4 (E4) with SCO explants from E4 or E13 embryos have been performed and the neurite outgrowth from the MNPCr explants has been analysed. In the case of coculture of E4 MNPCr with E4 SCO, the number of neurites growing from the MNPCr is higher at the side facing the SCO. However, when E4 MNPCr and E13 SCO are cocultured, the neurites grow mostly at the side opposite to the SCO. These data suggest that, at early stages of development, the SCO releases some attractive or permissive molecule(s) for the growing of the PC, whereas at later stages, the SCO has a repulsive effect over neurites arising from MNPCr.

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Tissue culture of bovine subcommissural organ

Cited by 13 publications

References 38 publications

Effects of neuroactive substances on the activity of subcommissural organ cells in dispersed cell and explant cultures

Effects of neuroactive substances on the activity of subcommissural organ cells in dispersed cell and explant cultures

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

The subcommissural organ and the development of the posterior commissure in chick embryos

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