Abstract
Here we describe the use of confocal microscopy in combination with antibodies specific to Golgi proteins to visualize dendritic Golgi outposts (GOPs) in cultured hippocampal pyramidal neurons. We also describe the use of spinning disk confocal microscopy, in combination with ectopically expressed glycosyltransferases fused to GFP variants, to visualize GOPs in living neurons.
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References
Kaech S, Banker G (2006) Culturing hippocampal neurons. Nat Protoc 1:2406–2415
Conde C, Cáceres A (2009) Microtubule assembly, organization and dynamics in axons and dendrites. Nat Rev Neurosci 10:319–332
Namba T, Funayashi Y, Nakamuta S, Xu C, Takano T, Kaibuchi K (2015) Extracellular and intracellular signalling for neuronal polarity. Physiol Rev 95:995–1024
Horton AC, Ehlers M (2003) Neuronal polarity and trafficking. Neuron 40:277–295
Ehlers MD (2013) Dendritic trafficking for neuronal growth and plasticity. Biochem Soc Trans 41:1365–1382
Horton AC, Rácz B, Monson EE, Lin AL, Weinberg RJ, Ehlers MD (2005) Polarized secretory trafficking directs cargo for asymmetric dendrite growth and morphogenesis. Neuron 48:751–771
Ye B, Zhang Y, Song W, Younger SH, Jan LY, Jan YN (2007) Growing dendrites and axons differ in their reliance on the secretory pathway. Cell 130:717–729
Quassollo G, Wojnacki J, Salas DA, Gastaldi L, Marzolo MP, Conde C, Bisbal M, Couve A, Cáceres A (2015) A RhoA signaling pathway regulates dendritic Golgi outpost formation. Curr Biol 25:971–982
Kaech S, Huang CF, Banker G (2012) General considerations for live imaging of developing hippocampal neurons in culture. Cold Spring Harb Protoc 201:312–318
Maccioni HFC, Quiroga R, Spessott W (2011) Organization of the synthesis of glycolipid oligosaccharides in the Golgi complex. FEBS Lett 585:1691–1698
Giraudo CG, Rosales Fritz VM, Maccioni HF (1999) Ga2/Gm2/Gd2 synthase localizes to the trans-Golgi network of cho-k1 cells. Biochem J 342:633–640
Giraudo C, Daniotti J, Maccioni H (2001) Physical and functional association of glycolipid N-acetyl-galactosaminyl and galactosyl transferases in the Golgi apparatus. Proc Natl Acad Sci U S A 98:1625–1630
Eliceiri KW, Berthold MR, Goldberg IG, Ibáñez L, Manjunath BS, Martone ME, Murphy RF, Peng H, Plant AL, Roysam B, Stuurman N, Swedlow JR, Tomancak P, Carpenter AE (2012) Biological imaging software tools. Nat Methods 9:697–710
Acknowledgements
Research at the Laboratory of Neurobiology at INIMEC-CONICET-UNC is supported by grants from FONCyT, CONICET to A.C. M.B. is supported by a return home grant from ISN and FONCyT grant (PICT D 2013-1525). A.C. and M.B. are staff scientists from CONICET.
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Bisbal, M., Quassollo, G., Caceres, A. (2016). Imaging Golgi Outposts in Fixed and Living Neurons. In: Brown, W. (eds) The Golgi Complex. Methods in Molecular Biology, vol 1496. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6463-5_3
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DOI: https://doi.org/10.1007/978-1-4939-6463-5_3
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