Regulation of the biosynthesis and processing of chromogranins in organotypic slices: influence of depolarization, forskolin and differentiating factors

Bauer, Richard; Hoflehner, Johannes; Doblinger, Alfred; Kapeller, I.; Laslop, Andrea

doi:10.1046/j.1460-9568.2000.00157.x

Cited by 8 publications

(4 citation statements)

References 74 publications

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“…2000). This is in line with a previous study on hippocampal organotypic slices showing also an increased expression for sg II but not for chromogranin B (Bauer et al . 2000b).…”

Section: Discussionsupporting

confidence: 93%

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Differential effects of phencyclidine application on secretogranin II expression in organotypic slices of rat prefrontal cortex

Hinterhoelzl

Salimi

Humpel

et al. 2003

Journal of Neurochemistry

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Phencyclidine (PCP) is a non-competitive NMDA glutamate receptor antagonist that induces psychotomimetic effects in humans and experimental animals. Chronic PCP exposure elicits signs of persistently altered frontal brain activity and related behaviors which are also seen in patients with schizophrenia. Secretogranin II (sg II) belongs to the chromogranin family of proteins that exist in large dense core vesicles in nervous tissue. In the brain, 90% of sg II is processed to the small peptide secretoneurin. We previously detected differential effects of single-dose and subchronic PCP administration on sg II expression in the rat prefrontal cortex (PFC). In the present study, we applied PCP to organotypic PFC slices. PCP application for 28 h induced decreased tissue and culture medium secretoneurin content. In contrast, incubation with the adenylate cyclase activator forskolin caused significantly increased secretoneurin levels after 8 h. PCP for 4 h followed by 24 h without PCP resulted in increased culture medium secretoneurin content but no change in tissue levels. sg II mRNA expression was decreased after 28 h PCP application in cortical neurons. Immunohistochemical and TUNEL staining profiles indicated that the alterations were not due to neurodegeneration. PCP for 5 days changed neither the secretoneurin tissue or culture medium levels, nor the sg II mRNA expression. These results demonstrate that PCP modulates sg II expression in PFC tissue in the absence of afferent inputs and that the nature of these changes is dependent upon the duration of exposure to and/or withdrawal from PCP. Keywords: gene expression, in vitro, neuropeptides, NMDA antagonist, prefrontal cortex, schizophrenia. J. Neurochem. (2003) 87, 13-21. Noncompetitive antagonists of the NMDA glutamate receptor, including the dissociative anesthetic phencyclidine (PCP), cause a psychotic reaction in humans comprising many of the positive and negative symptoms of schizophrenia (Tamminga 1998). Chronic PCP exposure elicits a schizophrenia-like pattern of impairment in social and cognitive performance that is thought to result from disturbed prefrontal cortical function (Andreasen et al. 1997;Bunney and Bunney 2000). NMDA glutamate receptor antagonism exerts effects on prefrontal cortical metabolism and neurotransmitter tone the quality and duration of which varies considerably depending on dosing regimen (for review, see Jentsch and Roth 1999). Specifically, chronic PCP exposure induces lasting cognitive impairment (Jentsch et al. 2000) as well as reduced frontal blood flow in humans (Hertzmann et al. 1990) and persistent deficits in prefrontal cortical dopamine utilization and prefrontal cortex (PFC)-dependent cognitive function in rats and monkeys (Balla et al. 2001).The chromogranins comprise a group of acidic secretory proteins which are stored with other neuropeptides in secretory large dense core vesicles (Winkler and Fischer-Colbrie 1992). Chromogranins are involved in the sorting, processing, Received April 23, 2003; accepted May...

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“…2000). This is in line with a previous study on hippocampal organotypic slices showing also an increased expression for sg II but not for chromogranin B (Bauer et al . 2000b).…”

Section: Discussionsupporting

confidence: 93%

“…Organotypic cultures of PFC were established as previously described for slices from other brain areas (Bauer et al . 2000b).…”

Section: Organotypic Prefrontal Cortical Slice Culturesmentioning

confidence: 99%

Differential effects of phencyclidine application on secretogranin II expression in organotypic slices of rat prefrontal cortex

Hinterhoelzl

Salimi

Humpel

et al. 2003

Journal of Neurochemistry

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“…However, the fact that exposure of L6 myoblasts to hypoxia without serum failed to increase pro‐SN mRNA (in contrast to VEGF) indicates that additional factors are required for pro‐SN up‐regulation. We hypothesized that bFGF might mediate an increase of SN by hypoxia, because bFGF is known to regulate pro‐SN (35) and to exert enhanced action in hypoxia by a HIF‐1α‐dependent increase of low‐affinity heparan sulfate bFGF binding sites (6). Several observations indicate that SN regulation by hypoxia in myoblasts is indeed mediated by bFGF.…”

Section: Discussionmentioning

confidence: 99%

Hypoxia up‐regulates the angiogenic cytokine secretoneurinviaan HIF‐1α‐ and basic FGF‐dependent pathway in muscle cells

Egger¹,

Schgoer²,

Beer³

et al. 2007

FASEB j.

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Expression of angiogenic cytokines like vascular endothelial growth factor is enhanced by hypoxia. We tested the hypothesis that decreased oxygen levels up-regulate the angiogenic factor secretoneurin. In vivo, muscle cells of mouse ischemic hind limbs showed increased secretoneurin expression, and inhibition of secretoneurin by a neutralizing antibody impaired the angiogenic response in this ischemia model. In a mouse soft tissue model of hypoxia, secretoneurin was increased in subcutaneous muscle fibers. In vitro, secretoneurin mRNA and protein were up-regulated in L6 myoblast cells after exposure to low oxygen levels. The hypoxia-dependent regulation of secretoneurin was tissue specific and was not observed in endothelial cells, vascular smooth muscle cells, or AtT20 pituitary tumor cells. The hypoxia-dependent induction of secretoneurin in L6 myoblasts is regulated by hypoxia-inducible factor-1alpha, since inhibition of this factor using si-RNA inhibited up-regulation of secretoneurin. Induction of secretoneurin by hypoxia was dependent on basic fibroblast growth factor in vivo and in vitro, and inhibition of this regulation by heparinase suggests an involvement of low-affinity basic fibroblast growth factor binding sites. In summary, our data show that the angiogenic cytokine secretoneurin is up-regulated by hypoxia in muscle cells by hypoxia-inducible factor-1alpha- and basic fibroblast growth factor-dependent mechanisms.

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“…The anti-CgA antibodies employed were the rabbit GE-19 antiserum (Bauer et al, 2000) and the mouse monoclonal LK2H10 (NeoMarkers, Fremont, CA, USA); the anti-CgBs were the rabbit PE11 antiserum (Kroesen et al, 1996) and the mouse monoclonal CIRO raised in our laboratory. The latter was characterised by western blotting and immunofluorescence and found to be highly specific.…”

Section: Antibodiesmentioning

confidence: 99%

Dense-core granules: a specific hallmark of the neuronal/neurosecretory cell phenotype

Malosio

Giordano

Laslop

et al. 2004

Journal of Cell Science

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Expression of dense-core granules, a typical exocytic organelle, is widely believed to be controlled by coordinate gene expression mechanisms specific to neurones and neurosecretory cells. Recent studies in PC12 cells, however, have suggested the number of granules/cells depends on the levels of only one of their cargo proteins, chromogranin A, regulating the metabolism of the other proteins, and thus the composition of the organelles, by an on/off switch mechanism. In addition, transfection of chromogranin A was reported to induce appearance of dense-core granules in the non-neurosecretory fibroblasts of the CV-1 line. Here the role of chromogranin A has been reinvestigated using not the heterogeneous PC12 line but several clones isolated therefrom. In these clones, investigated as such or after transfection with chromogranin A antisense sequences, the ratio between chromogranin A and its secretory protein mate, chromogranin B, was not constant but highly and apparently randomly variable. Variability of the chromogranin A/chromogranin B ratio was seen by confocal immunofluorescence also among the cells of single clones and subclones and among the granules of single cells. Moreover, stable and transient transfections of chromogranin A in a PC12 clone characterised by a low number of dense-core granules (one fifth of the reference clone) failed to modify significantly the number of the organelles, despite the several-fold increase of the granin. Finally, in three types of non-neurosecretory cells (CV-1, adenocarcinoma TS/A and a clone of PC12 incompetent for secretion) the transfected chromogranin A accumulated mostly in the Golgi/transGolgi area and was released rapidly from resting cells (constitutive secretion) as revealed by both immunofluorescence during cycloheximide treatment and pulse-chase experiments. Only a minor fraction was sorted to discrete organelles that were not dense-core granules, but primarily lysosomes because they contained no chromogranin B, and were largely positive for the late endosomal-lysosomal markers, lamp1 and lamp3. Dense-core granules are therefore true hallmarks of neurones and neurosecretory cells. Their number/cell appears independent of chromogranin A and their composition does not appear to be constant; in particular, they exhibit considerable, and so far unexplained variability in the chromogranin A/chromogranin B ratio.

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Regulation of the biosynthesis and processing of chromogranins in organotypic slices: influence of depolarization, forskolin and differentiating factors

Cited by 8 publications

References 74 publications

Differential effects of phencyclidine application on secretogranin II expression in organotypic slices of rat prefrontal cortex

Differential effects of phencyclidine application on secretogranin II expression in organotypic slices of rat prefrontal cortex

Hypoxia up‐regulates the angiogenic cytokine secretoneurinviaan HIF‐1α‐ and basic FGF‐dependent pathway in muscle cells

Dense-core granules: a specific hallmark of the neuronal/neurosecretory cell phenotype

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