Abstract
Recently, it has been proposed that neurofibromin (NF1) forms a binding complex with amyloid precursor protein (APP) that interacts with the dopamine D3 receptor (D3R). In the present study we investigated whether the absence of the D3R is correlated to modifications in the expression of both NF1 and APP. Quantitative real-time PCR analyses of both transcripts showed that NF1 mRNA levels were significantly reduced whereas APP levels were strikingly increased in D3R knock-out (D3R KO) as compared to wild type (WT) mice brains. Western blot analyses using mice whole brains produced comparable results with those obtained by mRNA measurements. Moreover, immunohistochemical analyses revealed a similar brain regional distribution of APP protein in the hippocampus, in the cerebral and cerebellar cortex of D3R KO mice. Conversely, hippocampal NF1 immunoreactivity did not seem to be affected by the absence of D3Rs. Further analyses confirmed that regional NF1 protein expression in the hippocampus was not affected by the absence of the D3R, whereas APP levels were still increased in this specific brain region. In conclusion, these results show the existence of a correlation among the D3R, NF1 and APP in mice brains and thus show the regional-specific regulation of NF1 in brains of D3R KO, which may contribute to gain insights into the comprehension of novel underlying mechanisms that regulate brain function.
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References
Missale C, Nash SR, Robinson SW et al (1998) Dopamine receptors: from structure to function. Physiol Rev 78:189–225
Sokoloff P, Giros B, Martres MP, Bouthenet ML et al (1990) Molecular cloning and characterization of a novel dopamine receptor (D3) as a target for neuroleptics. Nature 347:146–151
Donarum EA, Halperin RF, Stephan DA et al (2006) Cognitive dysfunction in NF1 knock-out mice may result from altered vesicular trafficking of APP/DRD3 complex. BMC Neurosci 7:20–31
Ozonoff S (1999) Cognitive impairment in neurofibromatosis type I. Am J Med Genet 89:45–52
Feldkamp MM, Angelov L, Guha A (1999) Neurofibromatosis type I peripheral nerve tumors: abberant activation of the Ras pathway. Surg Neurol 51:211–218
Costa RM, Federov NB, Kogan JH et al (2002) Mechanism for the learning deficits in a mouse model of neurofibromatosis type 1. Nature 415:526–530
Park CS, Zhong L, Tang SJ (2009) Aberrant expression of synaptic plasticity-related genes in the NF1 +/− mouse hippocampus. J Neurosci Res 87:3107–3119
Suh YH, Checler F (2002) Amyloid precursor protein, presenilins and alpha-synuclein: molecular pathogenesis and pharmacological applications in Alzheimer disease. Pharmacol Rev 54:469–525
Marcello E, Epis R, Di Luca M (2008) Amyloid flirting with synaptic failure: towards a comprehensive view of Alzheimer disease pathogenesis. Eur J Pharmacol 585:109–118
Rossjohn J, Cappai R, Feil SC et al (1999) Crystal structure of the N-terminal, growth factor-like domain of Alzheimer amyloid precursor protein. Nat Struct Biol 6:327–331
Perez RG, Zheng H, Van der Ploeg LH et al (1997) The beta-amyloid precursor protein of Alzheimer’s disease enhances neuron viability and modulates neuronal polarity. J Neurosci 17:9407–9414
Dawson GR, Seabrook H, Zheng DW et al (1999) Age-related cognitive deficits, impaired long-term potentiation and reduction in synaptic marker density in mice lacking the beta-amyloid precursor protein. Neuroscience 90:1–13
Seabrook GR, Rosahl TW (1999) Transgenic animals relevant to Alzheimer’s disease. Neuropharmacology 38:1–17
Koo EH, Sisodia SS, Archer DR et al (1990) Precursor of amyloid protein in Alzheimer disease undergoes fast axonal transport. Proc Natl Acad Sci USA 87:1561–1565
Sisodia SS, Koo EH, Hoffman PN et al (1993) Identification and transport of full length APP in rat peripheral nervous system. J Neurosci 13:3136–3142
Accili D, Fishburn CS, Drago J et al (1996) A targeted mutation of the D3 dopamine receptor gene is associated with hyperactivity in mice. Proc Natl Acad Sci USA 93:1945–1949
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3:1101–1108
Carroll SL, Ratner N (2008) How does the Schwann cell lineage form tumors in NF1? Glia 56:1590–1605
Naves FJ, Calzada B, Cabal A et al (1994) Expression of beta-amyloid precursor protein (APP) in human dorsal root ganglia. Neurosci Lett 181:73–77
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Pascale A, Fortino I, Govoni S et al (1996) Functional impairment in protein kinase C by RACK1 (receptor for activated C kinase 1) deficiency in aged rat brain cortex. J Neurochem 67:2471–2477
Lansner A (2009) Associative memory models: from the cell-assembly theory to biophysically detailed cortex simulations. Trends Neurosci 32:178–186
Lee E, Son H (2009) Adult hippocampal neurogenesis and related neurotrophic factors. BMB Rep 42:239–244
Manto M (2008) The cerebellum, cerebellar disorders, and cerebellar research-two centuries of discoveries. Cerebellum 7:505–516
Bollag G, McCormick F, Clark R (1993) Characterization of full-length neurofibromin: tubulin inhibits Ras GAP activity. EMBO J 12:1923–1927
Gregory PE, Gutmann DH, Mitchell A et al (1993) Neurofibromatosis type 1 gene product (neurofibromin) associates with microtubules. Somat Cell Mol Genet 19:265–274
Xu H, Gutman DH (1997) Mutations in the GAP-related domain impair the ability of neurofibromin to associate with microtubules. Brain Res 759:149–152
Li C, Cheng Y, Gutman DA et al (2001) Differential localization of the neurofibromatosis (NF1) gene product, neurofibromin with the F-actin or microtubule cytoskeleton during differentiation of telencephalic neurons. Dev Brain Res 130:231–248
Hakimi MA, Speicher DW, Shiekhattar R (2002) The motor protein kinesin-1 links neurofibromin and merlin in a common cellular pathway of neurofibromatosis. J Biol Chem 277:36909–36912
Hsueh YP, Roberts AM, Volta M et al (2001) Bipartite interaction between neurofibromatosis type 1 protein (neurofibromin) and syndecan transmembrane heparan sulfate proteoglycans. J Neurosci 21:3764–3770
Herzog V, Kirfal G, Siemens C et al (2004) Biological roles of APP in the epidermis. Eur J Cell Biol 83:613–624
Kamal A, Alemnar-Queralt A, Leblanc JF et al (2000) Axonal transport of amyloid precursor protein is mediated by direct binding to the kinesin light chain subunit of kinesin. Neuron 28:449–459
De Schepper S, Boucneau JMA, Westbroek W et al (2006) Neurofibromatosis type 1 protein and amyloid precursor protein interact in normal human melanocytes and colocalize with melanosomes. J Invest Dermatol 626:653–659
Acknowledgments
These experiments were supported by the international PhD program in Neuropharmacology, University of Catania, Medical School. We thank Mr. P. Asero and Mrs. R. Gambino for their technical support.
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Castorina, A., Leggio, G.M., Giunta, S. et al. Neurofibromin and Amyloid Precursor Protein Expression in Dopamine D3 Receptor Knock-Out Mice Brains. Neurochem Res 36, 426–434 (2011). https://doi.org/10.1007/s11064-010-0359-0
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DOI: https://doi.org/10.1007/s11064-010-0359-0