Abstract
Activity-dependent neurotrophic factor (ADNF) is a novel, femtomolar-acting, glial-derived polypeptide (14 kDa) known to protect neurons from a variety of toxic insults. The active site for ADNF function is localized to a 9-amino-acid stretch (SALLRSIPA; ADNF-9). A few years later, a novel ADNF-9-like active peptide (NAPVSIPQ or NAP) was identified and shown to be expressed in the CNS and exhibit an activity profile similar to ADNF-9. Such studies suggest that ADNF-9 and NAP might function like other known neurotrophins and play a role in neural development and maintenance. The purpose of the present studies was to determine if ADNF-9 or NAP affects neurite outgrowth and synaptogenesis in rat hippocampal and cortical cultures. Using MAP2-FITC immunofluorescent labeling, we found that ADNF-9 and NAP promoted neurite outgrowth in a concentration-dependent manner, with maximal activity observed at femtomolar concentrations. Both peptides stimulated robust outgrowth in hippocampal cells (∼150% of control; p<0.01) with a modest effect on cortical cells (∼20% of control; p<0.05)—similar to other known growth factors. However, the outgrowth-promoting effect was abolished in the absence of serum, suggesting that soluble factors might be necessary for the neurotrophic activity. Finally, we found that ADNF-9 and NAP increased synaptophysin expression in both rat hippocampal and cortical cultures. These results suggest that ADNF-9 and NAP might contribute to neuronal plasticity associated with development and repair after injury.
Similar content being viewed by others
References
Ashur-Fabian O., Giladi E., Furman S., Steingart R. A., Wollman Y., Fridkin M., et al. (2001) Vasoactive intestinal peptide and related molecules induce nitrite accumulation in the extracellular milieu of rat cerebral cortical cultures. Neurosci. Lett. 307, 167–170.
Bassan M., Zamostiano R., Davidson A., Pinhasov A., Giladi E., Perl O., et al. (1999) Complete sequence of a novel protein containing a femtomolar-activity-dependent neuroprotective peptide. J. Neurochem. 72, 1283–1293.
Bassan M., Zamostiano R., Giladi E., Davidson A., Wollman Y., Pitman J., et al. (1998) The identification of secreted heat shock 60-like protein from rat glial cells and a human neuroblastoma cell line. Neurosci. Lett. 249, 1–4.
Beni-Adani L., Gozes I., Cohen Y., Assaf Y., Steingart R. A., Brenneman D. E., et al. (2001) A peptide derived from activity-dependent neuroprotective protein (ADNP) ameliorates injury response in closed head injury mice. J. Pharmacol. Exp. Ther. 296, 57–63.
Blondel O., Collin C., McCarran B., Zhu S., Zamostiano R., Gozes I., et al. (2000) A glia-derived signal regulating neuronal differentiation. J. Neurosci. 20, 8012–8020.
Brenneman D. E. and Eiden L. E. (1986). Vasoactive intestinal peptide and electrical activity influence neuronal survival. Proc. Natl. Acad. Sci. U. S. A. 83, 1159–1162.
Brenneman D. E. and Gozes I. (1996) A femtomolar-acting neuroprotective peptide. J. Clin. Invest. 97, 2299–2307.
Brenneman D. E., Hauser J., Neale E. A., Rubinraut S., Fridkin M., Davidson A., and Gozes I. (1998) Activity-dependent neurotrophic factor: structure-activity relationships of femtomolar-acting peptides. J. Pharmacol. Exp. Ther. 285, 619–627.
Brenneman D. E., Hauser J., Philips T. M., Davidson A., Bassan M., and Gozes I. (1999) Vasoactive intestinal peptide. Link between electrical activity and glia-mediated neurotrophism. Ann. N. Y. Acad. Sci. 897, 17–26.
Brenneman D. E., Phillips T. M., Festoff B. W., and Gozes I. (1997) Identity of neurotrophic molecules released from astroglia by vasoactive intestinal peptide. Ann. N. Y. Acad. Sci. 814, 167–173.
Canossa M., Griesbeck O., Berninger B, Campana G., Kolbeck R., and Thoenen H. (1997) Neurotrophin release by neurotrophins: implications for activity-dependent neuronal plasticity. Proc. Natl. Acad. Sci. U. S. A. 94, 13279–13286.
Das K. P., Chao S. L., White L. D., Haines W. T., Harry G. J., Tilson H. A., and Barrone S., Jr. (2001) Differential patterns of nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3 mRNA and protein levels in developing regions of rat brain. Neuroscience 103, 739–761.
Divinski I., Mittelman L., and Gozes I. (2004) A femtomolar acting octapeptide interacts with tubulin and protects astrocytes against zinc intoxication. J. Biol. Chem. 279, 28531–28538.
Eilam R., Davidson A., Gozes I., and Segal M. (1999) Locomotor activity causes a rapid up-regulation of vasoactive intestinal peptide in the rat hippocampus. Hippocampus 9, 534–541.
Glazner G. W., Boland A., Dresse A. E., Brenneman D. E., Gozes I., and Mattson M. P. (1999a) Activity-dependent neurotrophic factor peptide (ADNF9) protects neurons against oxidative stress-induced death. J. Neurochem. 73, 2341–2347.
Glazner G. W., Camandola S., and Mattson M. P. (2000) Nuclear factor-kappa B mediates the cell survival-promoting action of activity-dependent neurotrophic factor peptide-9. J. Neurochem. 75, 101–108.
Glazner G. W., Gressens P., Lee S. J., Gibney I., Gozes I., Brenneman D. E., and Hill J. M. (1999b) Activity-dependent neurotrophic factor: a potent regulator of embryonic growth and development. Anat. Embryol. 200, 65–71.
Gozes I. and Brenneman D. E. (1996) Activity-dependent neurotrophic factor ADNF: an extracellular neuroprotective chaperonin? J. Mol. Neurosci. 7, 235–244.
Gozes I. and Brenneman D. E. (2000) A new concept in neuroprotection. J. Mol. Neurosci. 14, 61–68.
Gozes I., Bassan M., Zamostiano R., Pinhasov A., Davidson A., Giladi E., et al. (1999) A novel signaling molecule for neuropeptide action: activity-dependent neuroprotective protein. Ann. N. Y. Acad. Sci. 897, 125–135.
Gozes I., Davidson A., Gozes Y., Mascolo R., Barth R., Warren D., Hauser J., and Brenneman D. E. (1997) Antiserum to activity-dependent neurotrophic factor produces neuronal cell death in CNS cultures: immunological and biological specificity. Dev. Brain Res. 99, 167–175.
Gozes I., Giladi E., Pinhasov A., Bardea A. and Brenneman D. E. (2000) Activity-dependent neurotrophic factor: intranasal administration of femtomolar-acting peptides improve performance in a watermaze. J. Pharmacol. Exp. Ther. 293, 1091–1098.
Gozes I., McCune S. K., Jacobson L., Warren D., Moody T. W., Fridkin M., and Brenneman D. E. (1991) An antagonist to vasoactive intestinal peptide: effects on cellular functions in the central nervous system. J. Pharmacol. Exp. Ther. 257, 959–966.
Gozes I., Shani Y., and Rostene W. H. (1987) Developmental expression of the VIP-gene in brain and intestine. Brain Res. 388, 137–148.
Gozes I., Steingart R. A., and Spier A. D. (2004) NAP mechanisms of neuroprotection. J. Mol. Neurosci. 24, 67–72.
Gressens P., Marret S., Bodenant C., Schwendimann L., and Evrard P. (1999) Activity-dependent neurotrophic factor-14 requires protein kinase C and mitogen-associated protein kinase kinase activation to protect the developing mouse brain against excitotoxicity. J. Mol. Neurosci. 13, 199–210.
Guo Q., Sebastian L., Sopher B. L., Miller M. W., Glazner G. W., Ware C. B., et al. (1999) Neurotrophic factors, activity-dependent neurotrophic factor (ADNF) and basic fibroblast growth factor (bFGF), interrupt excitotoxic neurodegenerative cascades promoted by a PS1 mutation. Proc. Natl. Acad. Sci. U. S. A. 96, 4125–4130.
Guo Z. H. and Mattson M. P. (2000) Neurotrophic factors protect cortical synaptic terminals against amyloid and oxidative stress-induced impairment of glucose transport, glutamate transport and mitochondrial function. Cereb. Cortex 10, 50–57.
Hill J. M., Lee S. J., Dibbern D. A., Fridkin M., Gozes I., and Brenneman D. E. (1999) Pharmacologically distinct vasoactive intestinal peptide binding sites: CNS localization and role in embryonic growth. Neuroscience 93, 783–791.
Kaiser P. K. and Lipton S. A. (1990) VIP-mediated increase in cAMP prevents tetrodotoxin-induced retinal ganglion cell death in vitro. Neuron 5, 373–381.
Kruttgen A., Moller J. C., Heymach J. V., and Shooter E. M. (1998) Neurotrophins induce release of neurotrophins by the regulated secretory pathway. Proc. Natl. Acad. Sci. U. S. A. 95, 9614–9619.
Leker R. R., Teichner A., Grigoriadis N., Ovadia H., Brenneman D. E., Fridkin M., et al. (2002) NAP, a femtomolar-acting peptide, protects the brain against ischemic injury reducing apoptotic death. Stroke 33, 1085–1092.
Mattson M. P., Barger S. W., Begley J. G., and Mark R. J. (1995) Calcium, free radicals, and excitotoxic neuronal death in primary cell culture. Methods Cell Biol. 46, 187–216.
Maxwell I. H., Glode L. M., and Maxwell F. (1992) Expression of diphtheria toxin A chain in mature B-cells, a potential approach to therapy of B-lymphoid malignancy. Leuk. Lymphoma 7, 457–462.
Offen D., Sherki Y., Melamed E., Fridkin M., Brenneman D. E., and Gozes I. (2000) Vasoactive intestinal peptide (VIP) prevents neurotoxicity in neuronal cultures: relevance to neuroprotection in Parkinson’s disease. Brain Res. 854, 257–262
Pfrieger F. W. and Barres B. A. (1997) Synaptic efficacy enhanced by glial cells in vitro. Science 277, 1684–1687.
Pincus D. W., DiCicco-Bloom E. M., and Black I. B. (1990) Vasoactive intestinal peptide regulates mitosis, differentiation and survival of cultured sympathetic neuroblasts. Nature 343, 564–567.
Pinhasov A., Mandel S., Torchinsky A., Giladi E., Pittel Z., Goldsweig A. M., et al. (2003) Activity-dependent neuroprotective protein: a novel gene essential for brain formation. Dev. Brain Res. 144, 83–90.
Poggi S. H., Vink J., Goodwin K., Hill J. M., Brenneman D. E., Pinhasov A., et al. (2002) Differential expression of embryonic and maternal activity-dependent neuroprotective protein during mouse development. Am. J. Obstet. Gynecol. 187, 973–976.
Smith-Swintosky V. L., Cheo-Isaacs C. T., D’Andrea M. R., Santulli R. J., Darrow A. L., and Andrade-Gordon P. (1997) Protease-activated receptor (PAR-2) is present in the rat hippocampus and is associated with neurodegeneration. J. Neurochem. 69, 1890–1896.
Soltys B. J. and Gupta R. S. (1999) Mitochondrial molecular chaperones hsp60 and mhasp70: are their roles restricted to mitochondria? Handbook Exp. Pharmacol. 136, 69–100.
Spong C. Y., Abebe D. T., Gozes I., Brenneman D. E., and Hill J. M. (2001) Prevention of fetal demise and growth restriction in a mouse model of fetal alcohol syndrome. J. Pharmacol. Exp. Ther. 297, 774–779.
Steingart R. A., Solomon B., Brenneman D. E., Fridkin M., and Gozes I. (2000) VIP and peptides related to activity-dependent neurotrophic factor protect PC12 cells against oxidative stress. J. Mol. Neurosci. 15, 137–145.
Vicario-Abejon C., Collin C., McKay R. D. G., and Segal M. (1998) Neurotrophins induce formation of functional excitatory and inhibitory synapses between cultured hippocampal neurons. J. Neurosci. 18, 7256–7271.
White D. M., Walker S., Brenneman D. E., and Gozes I. (2000) CREB contributes to the increased neurite out-growth of sensory neurons induced by vasoactive intestinal polypeptide and activity-dependent neurotrophic factor. Brain. Res. 868, 31–38.
Zamostiano R., Pinhasov A., Bassan M., Perl O., Steingart R. A., Atlas R., et al. (1999) A femtomolar-acting neuroprotective peptide induces increased levels of heat shock protein 60 in rat cortical neurons: a potential neuroprotective mechanism. Neurosci. Lett. 264, 9–12.
Zamostiano R., Pinhasov A., Gelber E., Steingart R. A., Seroussi E., Giladi E. et al. (2001) Cloning and characterization of the human activity-dependent neuroprotective protein. J. Biol. Chem. 276, 708–714
Zemlyak I., Furman S., Brenneman D. E., and Gozes I. (2000) A novel peptide (NAP) prevents death in enriched neuronal cultures. Regul. Pept. 96, 39–43.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Smith-Swintosky, V.L., Gozes, I., Brenneman, D.E. et al. Activity-dependent neurotrophic factor-9 and NAP promote neurite outgrowth in rat hippocampal and cortical cultures. J Mol Neurosci 25, 225–238 (2005). https://doi.org/10.1385/JMN:25:3:225
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1385/JMN:25:3:225