Abstract
The neurotrophin brain-derived neurotrophic factor (BDNF) is extensively expressed in neural cells and other cell types. BDNF displays a support function in both central CNS and peripheral PNS where it is involved in most neuronal mechanisms, from development to neurodegeneration, cellular growth, neuroprotection, survival, and network signaling. In the last decades, decreased levels of BDNF were directly linked to Parkinson’s, Alzheimer’s, and Huntington’s disease, while BDNF upregulation was found in patients affected by epilepsy, autism, and multiple sclerosis. Accordingly, determination of BDNF expression can contribute to gain insight in several pathological models both in vivo and in vitro. In this manuscript, we provide detailed description of the main techniques for BDNF detection in cultured neurons and in brain slices, using different methods such as ELISA, Western blot, and immunofluorescence.
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References
Barde YA, Edgar D, Thoenen H (1982) Purification of a new neurotrophic factor from mammalian brain. EMBO J 1(5):549–553
Mizui T, Washikawa Y, Kumanogoh H, Lume M, Matsumoto T, Hara T, Yamawaki S, Takahashi M, Shiosaka S, Itami C, Uega ki K, Saarma M, Kojima M (2015) BDNF pro-peptide actions facilitate hippocampal LTD and are altered by the common BDNF polymorphism Val66Met. Proc Natl Acad Sci U S A 112(23):E3067–E3074
Ibanez CF (1998) Emerging themes in structural biology of neurotrophic factors. Trends Neurosci 21:438–444
Mowla SJ, Farhadi HF, Pareek S, Atwal JK, Morris SJ, Seidah NG, Murphy RA (2001) Biosynthesis and post-translational processing of the precursor to brain-derived neurotrophic factor. J Biol Chem 276(16):12660–12666
Lee R, Kermani P, Teng KK, Hempstead BL (2001) Regulation of cell survival by secreted proneurotrophins. Science 294(5548):1945–1948
Seidah NG, Mowla SJ, Hamelin J, Mamarbachi AM, Benjannet S, Touré BB, Basak A, Munzer JS, Marcinkiewicz J, Zhong M, Barale JC, Lazure C, Murphy RA, Chrétien M, Marcinkiewicz M (1999) Mammalian subtilisin/kexin isozyme SKI-1: a widely expressed proprotein convertase with a unique cleavage specificity and cellular localization. Proc Natl Acad Sci U S A 96(4):1321–1326
Greenberg ME, Xu B, Lu B, Hempstead BL (2009) New insights in the biology of BDNF synthesis and release: implications in CNS function. J Neurosci 29(41):12764–12767
Koshimizu H, Kiyosue K, Hara T, Hazama S, Suzuki S, Uegaki K, Nagappan G, Zaitsev E, Hirokawa T, Tatsu Y, Ogura A, Lu B, Kojima M (2009) Multiple functions of precursor BDNF to CNS neurons: negative regulation of neurite growth, spine formation and cell survival. Mol Brain 2:27
Teng HK, Teng KK, Lee R, Wright S, Tevar S, Almeida RD, Kermani P, Torkin R, Chen ZY, Lee FS, Kraemer RT, Nykjaer A, Hempstead BL (2005) ProBDNF induces neuronal apoptosis via activation of a receptor complex of p75NTR and sortilin. J Neurosci 25(22):5455–5463
Woo NH, Teng HK, Siao CJ, Chiaruttini C, Pang PT, Milner TA, Hempstead BL, Lu B (2005) Activation of p75NTR by proBDNF facilitates hippocampal long-term depression. Nat Neurosci 8(8):1069–1077
Sermasi E, Margotti E, Cattaneo A, Domenici L (2000) Trk B signalling controls LTP but not LTD expression in the developing rat visual cortex. Eur J Neurosci 12(4):1411–1419
Nagappan G, Zaitsev E, Senatorov VV Jr, Yang J, Hempstead BL, Lu B (2009) Control of extracellular cleavage of ProBDNF by high frequency neuronal activity. Proc Natl Acad Sci U S A 106(4):1267–1272
Barnes P, Thomas KL (2008) Proteolysis of proBDNF is a key regulator in the formation of memory. PLoS One 3(9):e3248
Mizui T, Ohira K, Kojima M (2017) BDNF pro-peptide: a novel synaptic modulator generated as an N-terminal fragment from the BDNF precursor by proteolytic processing. Neural Regen Res 12(7):1024–1027
Carlino D, Leone E, Di Cola F, Baj G, Marin R, Dinelli G, Tongiorgi E, De Vanna M (2011) Low serum truncated-BDNF isoform correlates with higher cognitive impairment in schizophrenia. J Psychiatr Res 45(2):273–279
Garcia KL, Yu G, Nicolini C, Michalski B, Garzon DJ, Chiu VS, Tongiorgi E, Szatmari P, Fahnestock M (2012) Altered balance of proteolytic isoforms of pro-brain-derived neurotrophic factor in autism. J Neuropathol Exp Neurol 71(4):289–297
Carlino D, De Vanna M, Tongiorgi E (2012) Is altered BDNF biosynthesis a general feature in patients with cognitive dysfunctions? Neuroscientist 19(4):345–353
Aid T, Kazantseva A, Piirsoo M, Palm K, Timmusk T (2007) Mouse and rat BDNF gene structure and expression revisited. J Neurosci Res 85(3):525–535
Pruunsild P, Kazantseva A, Aid T, Palm K, Timmusk T (2007) Dissecting the human BDNF locus: bidirectional transcription, complex splicing, and multiple promoters. Genomics 90(3):397–406
Matsuda N, Lu H, Fukata Y, Noritake J, Gao H, Mukherjee S, Nemoto T, Fukata M, Poo MM (2009) Differential activity-dependent secretion of brain-derived neurotrophic factor from axon and dendrite. J Neurosci 29(45):14185–14198
Dieni S, Matsumoto T, Dekkers M, Rauskolb S, Ionescu MS, Deogracias R, Gundelfinger ED, Kojima M, Nestel S, Frotscher M, Barde YA (2012) BDNF and its pro-peptide are stored in presynaptic dense core vesicles in brain neurons. J Cell Biol 196(6):775–788
Simonato M, Bregola G, Armellin M, Del Piccolo P, Rodi D, Zucchini S, Tongiorgi E (2002) Dendritic targeting of mRNAs for plasticity genes in experimental models of temporal lobe epilepsy. Epilepsia 43(Suppl 5):153–158
Tongiorgi E, Armellin M, Giulianini PG, Bregola G, Zucchini S, Paradiso B, Steward O, Cattaneo A, Simonato M (2004) Brain-derived neurotrophic factor mRNA and protein are targeted to discrete dendritic laminas by events that trigger epileptogenesis. J Neurosci 24(30):6842–6852
Edelmann E, Lessmann V, Brigadski T (2014) Pre- and postsynaptic twists in BDNF secretion and action in synaptic plasticity. Neuropharmacology 76 Pt C:610–627
Polacchini A, Metelli G, Francavilla R, Baj G, Florean M, Mascaretti LG, Tongiorgi E (2015) A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays. Sci Rep 5:17989
Baj G, Patrizio A, Montalbano A, Sciancalepore M, Tongiorgi E (2014) Developmental and maintenance defects in Rett syndrome neurons identified by a new mouse staging system in vitro. Front Cell Neurosci 8:18
Acknowledgments
The authors thank Dr. Elsa Fabbretti for critically reading the manuscript. This work has been supported by the University of Trieste internal funds FRA 2016 to E.T.
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Donegà, S., Tongiorgi, E. (2018). Detecting BDNF Protein Forms by ELISA, Western Blot, and Immunofluorescence. In: Duarte, C., Tongiorgi, E. (eds) Brain-Derived Neurotrophic Factor (BDNF). Neuromethods, vol 143. Humana, New York, NY. https://doi.org/10.1007/7657_2018_13
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DOI: https://doi.org/10.1007/7657_2018_13
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