Peptide repertoire of human cerebrospinal fluid: novel proteolytic fragments of neuroendocrine proteins

Stark, Margareta; Danielsson, Olle; Griffiths, William J.; Jörnvall, Hans; Johansson, Jan

doi:10.1016/s0378-4347(00)00628-9

Cited by 69 publications

(70 citation statements)

References 62 publications

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“…They are processed from protein precursors by a proteolytic mechanism, packaged into large dense core vesicles, and released to bind receptors on the postsynaptic target neuron. Our peptidome data set shows that these peptides are all processed at dibasic sites or before single Arg residues as previously reported for polypeptides detected in cerebrospinal fluid (43). In our peptidome, we evidenced new processed and secreted neuropeptides: VGF peptide 24 -63, SCGI peptide 304 -321; and PENK peptides 114 -133, 143-185, and 239 -260.…”

Section: Discussionsupporting

confidence: 83%

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Discovering New Bioactive Neuropeptides in the Striatum Secretome Using in Vivo Microdialysis and Versatile Proteomics

Bernay

Gaillard

Guryča

et al. 2009

Molecular & Cellular Proteomics

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The striatum, a major component of the brain basal nuclei, is central for planning and executing voluntary movements and undergoes lesions in neurodegenerative disorders such as Huntington disease. To perform highly integrated tasks, the striatum relies on a complex network of communication within and between brain regions with a key role devoted to secreted molecules. To characterize the rat striatum secretome, we combined in vivo microdialysis together with proteomics analysis of trypsin digests and peptidomics studies of native fragments. This versatile approach, carried out using different microdialysis probes and mass spectrometer devices, allowed evidencing with high confidence the expression of 88 proteins and 100 processed peptides. Their secretory pathways were predicted by in silico analysis. Whereas high molecular weight proteins were mainly secreted by the classical mode (94%), low molecular weight proteins equally used classical and non-classical modes (53 and 47%, respectively). In addition, our results suggested alternative secretion mechanisms not predicted by bioinformatics tools. Based on spectrum counting, we performed a relative quantification of secreted proteins and peptides in both basal and neuronal depolarization conditions. This allowed detecting a series of neuropeptide precursors and a 6-fold increase for neurosecretory protein VGF and In mammalian brain, the striatum plays a critical role for planning and executing voluntary movements and is also involved in cognitive processes (1). The striatum makes use of a complex network architecture connecting specialized anatomical structures to achieve these highly integrated tasks. It receives projections from primary sensory and motor cortices as well as motor thalamic nuclei and sends projections to downstream basal ganglia structures, thereby influencing the control of cortical and brainstem motor systems (2). In this context, communication within and between brain structures appears as a key element for brain functioning. For cell-to-cell communication, secreted proteins play a pivotal regulatory role. To enter the secretory pathway, it has been long assumed that an N-terminal signal peptide sequence is strictly required. However, recent studies have shown that endoplasmic reticulum-and Golgi-independent or non-classical mechanisms may be responsible for protein secretion (3). The extracellular medium is thus more complex than previously suspected, and its characterization has gained a special interest (4, 5). In silico analyses suggest that mature proteins secreted via classical and non-classical mechanisms share common physicochemical properties (6). In this respect, proteomics is a powerful approach for systematically analyzing proteins present in the extracellular medium (7-9). For neurochemical monitoring of the secretome within the brain, only a few tools provide an appropriate insight into its spatial and temporal dynamics. Microdialysis, in particular, has been shown to be a powerful tool for exploring the extracellular content o...

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Section: Discussionsupporting

confidence: 83%

“…One characteristic feature of these peptides is a very high content of acidic residues, a hallmark for secreted peptides (43,47,48). The data obtained in the experiments with KCl, along with previous data (15), definitively demonstrate that all PENK fragments are co-secreted in the striatum.…”

Section: Discussionsupporting

confidence: 69%

Discovering New Bioactive Neuropeptides in the Striatum Secretome Using in Vivo Microdialysis and Versatile Proteomics

Bernay

Gaillard

Guryča

et al. 2009

Molecular & Cellular Proteomics

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“…However, we did not know if Complement could be activated during postnatal cerebellar development by either mechanism of activation. Nevertheless, peptide fragments generated by proteolysis at dibasic sites have been described in the cerebrospinal fluid, suggesting an action of pro-hormone convertases (54). Thus, C4a anaphylatoxin was found in the cerebrospinal fluid, and we can suppose that C3a and C5a could be cleaved from C3 and C5 in brain during development by a similar pro-hormone convertase mechanism.…”

Section: Discussionmentioning

confidence: 92%

Characterization of C3a and C5a Receptors in Rat Cerebellar Granule Neurons during Maturation

Bénard

Gonzalez

Schouft

et al. 2004

Journal of Biological Chemistry

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There is now clear evidence that the Complement anaphylatoxin C3a and C5a receptors (C3aR and C5aR) are expressed in glial cells, notably in astrocytes and microglia. In contrast, very few data are available concerning the possible expression of these receptors in neurons. Here, we show that transient expression of C3aR and C5aR occurs in cerebellar granule neurons in vivo with a maximal density in 12-day-old rat, suggesting a role of these receptors during development of the cerebellum. Expression of C3aR and C5aR mRNAs and proteins was also observed in vitro in cultured cerebellar granule cells. Quantification of the mRNAs by real-time reverse transcription-PCR showed a peak of expression at day 2 in vitro (DIV 2); the C3aR and C5aR proteins were detected by Western blot analysis at DIV 4 and by flow cytometry and immunocytochemistry in differentiating neurons with a maximum density at DIV 4 -9. Apoptosis of granule cells plays a crucial role for the harmonious development of the cerebellar cortex. We found that, in cultured granule neurons in which apoptosis was induced by serum deprivation and low potassium concentration, a C5aR agonist promoted cell survival and inhibited caspase-3 activation and DNA fragmentation. The neuroprotective effect of the C5aR agonist was associated with a marked inhibition of caspase-9 activity and partial restoration of mitochondrial integrity. Our results provide the first evidence that C3aR and C5aR are both expressed in cerebellar granule cells during development and that C5a, but not C3a, is a potent inhibitor of apoptotic cell death in cultured granule neurons.

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“…In addition, a few further VGF-derived peptides have been identified, but would not be recognized by the antisera we used. These include proVGF N-terminus and VGF 378-397 -related peptides identified from the human cerebrospinal fluid (Stark et al 2001, Carrette et al 2003. Furthermore, the sheep vgf gene has not yet been sequenced, hence its coding region might differ from the highly conserved sequence found in rat, mouse and human genes (Salton et al 2000).…”

Section: Discussionmentioning

confidence: 99%

Differential expression and seasonal modulation of VGF peptides in sheep pituitary

Brancia

Nicolussi²,

Cappai³

et al. 2005

Journal of Endocrinology

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The inducible gene vgf and its peptide products are relevant to the neuroendocrine regulation of homeostasis and reproduction in rodents. We show here that in the anterior pituitary of female sheep the somatotrope, gonadotrope, and lactotrope/thyrotrope cell populations each expressed vgf mRNA, but displayed a distinct profile of VGF immunoreactive peptides. ProVGF C-terminus and VGF 443-588 immunoreactivities were found in lactotropes and thyrotropes, often in a subcellular location restricted to the Golgi area and suggestive of rapid peptide (or proVGF) release upon biosynthesis, while high molecular weight bands consistent with proVGF were shown in pituitary extracts. Distinct seasonal changes were revealed, proVGF C-terminus immunoreactive cells being largely identified as lactotropes during the summer (83·7 2·1% (mean S.E.M.) versus 27·0 1·9% during the winter), as opposed to thyrotropes during the winter (73·0 1·9% versus 16·3 2·1% during the summer). Conversely, antisera to peptides adjacent to the VGF 553-555 'Arg-Pro-Arg' cleavage site, and to the N-terminus of the proVGF-derived peptide V, selectively labeled gonadotropes, indicating processing to small peptides not retaining the proVGF C-terminus in such cells. Finally, a peptide related to the VGF 4-240 region was immunostained in somatotropes, shown in a Western blot as a band of relative molecular mass of approximately 16 000. In conclusion, a complex, endocrine cell-typespecific processing of proVGF was revealed. Further to the known inducibility of vgf mRNA upon a range of stimuli, discreet, selective modulations of VGF-peptide profile/s are suggested, possibly involved in specific neuro/ endocrine or modulatory mechanisms.

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Peptide repertoire of human cerebrospinal fluid: novel proteolytic fragments of neuroendocrine proteins

Cited by 69 publications

References 62 publications

Discovering New Bioactive Neuropeptides in the Striatum Secretome Using in Vivo Microdialysis and Versatile Proteomics

Discovering New Bioactive Neuropeptides in the Striatum Secretome Using in Vivo Microdialysis and Versatile Proteomics

Characterization of C3a and C5a Receptors in Rat Cerebellar Granule Neurons during Maturation

Differential expression and seasonal modulation of VGF peptides in sheep pituitary

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