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Laminar Quantification of Dendrites in Dentate Gyrus Granule Neurons

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Multidisciplinary Tools for Investigating Synaptic Plasticity

Part of the book series: Neuromethods ((NM,volume 81))

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Abstract

Synaptic integration of young neurons requires the sprouting and extension of dendrites in order to establish contacts with afferent neurons. For this reason, dendritic growth is often used in developmental studies as an important indicator of neuronal health and maturity. In the hippocampus, new granule neurons are continuously produced throughout life. These neurons are initially arranged linearly along the inner border of the granule cell layer in the dentate gyrus and as they mature, their dendrites are extended perpendicularly into the molecular layer. Based on this known topography of the dentate gyrus, we outline here a method for analysis of dendritic growth in immature adult-born granule neurons, using laminar quantification of cell bodies along with primary, secondary, and tertiary dendrites separately and independently from each other. In contrast to other methods which often require the use of exogenous markers and/or arbitrary selection of individual neurons, laminar quantification of dendrites relies on immunohistochemical detection of endogenous markers to perform a comprehensive analysis of a subpopulation of immature neurons. The calculated parameters can be used in a comparative analysis to indicate variations in dendritic growth and complexity, thus providing important information regarding the development of young neurons in the adult hippocampus.

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Abbreviations

DCX:

Doublecortin

DG:

Dentate gyrus

GCL:

Granule cell layer

GFP:

Green fluorescent protein

ML:

Molecular layer

SGZ:

Sub-granular zone

References

  1. Sanes JR, Yamagata M (1999) Formation of lamina-specific synaptic connections. Curr Opin Neurobiol 9:79–87

    Article  PubMed  CAS  Google Scholar 

  2. Barinaga M (1995) Dendrites shed their dull image. Science 268:200–201

    Article  PubMed  CAS  Google Scholar 

  3. Spruston N, Schiller Y, Stuart G, Sakmann B (1995) Activity-dependent action potential invasion and calcium influx into hippocampal CA1 dendrites. Science 268:297–300

    Article  PubMed  CAS  Google Scholar 

  4. Bodnarenko SR, Jeyarasasingam G, Chalupa LM (1995) Development and regulation of dendritic stratification in retinal ganglion cells by glutamate-mediated afferent activity. J Neurosci 15:7037–7045

    PubMed  CAS  Google Scholar 

  5. Wingate RJ (1996) Retinal ganglion cell dendritic development and its control. Filling the gaps. Mol Neurobiol 12:133–144

    Article  PubMed  CAS  Google Scholar 

  6. Ruth RE, Collier TJ, Routtenberg A (1982) Topography between the entorhinal cortex and the dentate septotemporal axis in rats: I. Medial and intermediate entorhinal projecting cells. J Comp Neurol 209:69–78

    Article  PubMed  CAS  Google Scholar 

  7. Witter MP (2007) The perforant path: projections from the entorhinal cortex to the dentate gyrus. Prog Brain Res 163:43–61

    Article  PubMed  Google Scholar 

  8. Wang S, Scott BW, Wojtowicz JM (2000) Heterogeneous properties of dentate granule neurons in the adult rat. J Neurobiol 42: 248–257

    Article  PubMed  CAS  Google Scholar 

  9. Christie BR, Cameron HA (2006) Neurogenesis in the adult hippocampus. Hippocampus 16:199–207

    Article  PubMed  CAS  Google Scholar 

  10. Carlen M, Cassidy RM, Brismar H, Smith GA, Enquist LW, Frisen J (2002) Functional integration of adult-born neurons. Curr Biol 12:606–608

    Article  PubMed  CAS  Google Scholar 

  11. Abrous DN, Koehl M, Le Moal M (2005) Adult neurogenesis: from precursors to network and physiology. Physiol Rev 85: 523–569

    Article  PubMed  CAS  Google Scholar 

  12. Eadie BD, Redila VA, Christie BR (2005) Voluntary exercise alters the cytoarchitecture of the adult dentate gyrus by increasing cellular proliferation, dendritic complexity, and spine density. J Comp Neurol 486:39–47

    Article  PubMed  Google Scholar 

  13. Redila VA, Christie BR (2006) Exercise-induced changes in dendritic structure and complexity in the adult hippocampal dentate gyrus. Neuroscience 137:1299–1307

    Article  PubMed  CAS  Google Scholar 

  14. Rao MS, Hattiangady B, Abdel-Rahman A, Stanley DP, Shetty AK (2005) Newly born cells in the ageing dentate gyrus display normal migration, survival and neuronal fate choice but endure retarded early maturation. Eur J Neurosci 21:464–476

    Article  PubMed  Google Scholar 

  15. Ramirez-Rodriguez G, Ortiz-Lopez L, Dominguez-Alonso A, Benitez-King GA, Kempermann G (2011) Chronic treatment with melatonin stimulates dendrite maturation and complexity in adult hippocampal neurogenesis of mice. J Pineal Res 50:29–37

    Article  PubMed  CAS  Google Scholar 

  16. Tronel S, Fabre A, Charrier V, Oliet SH, Gage FH, Abrous DN (2010) Spatial learning sculpts the dendritic arbor of adult-born hippocampal neurons. Proc Natl Acad Sci U S A 107:7963–7968

    Article  PubMed  CAS  Google Scholar 

  17. Overstreet-Wadiche LS, Bromberg DA, Bensen AL, Westbrook GL (2006) Seizures accelerate functional integration of adult-generated granule cells. J Neurosci 26: 4095–4103

    Article  PubMed  CAS  Google Scholar 

  18. Zhao C, Teng EM, Summers RG Jr, Ming GL, Gage FH (2006) Distinct morphological stages of dentate granule neuron maturation in the adult mouse hippocampus. J Neurosci 26:3–11

    Article  PubMed  CAS  Google Scholar 

  19. Nishi M, Usuku T, Itose M, Fujikawa K, Hosokawa K, Matsuda KI, Kawata M (2007) Direct visualization of glucocorticoid receptor positive cells in the hippocampal regions using green fluorescent protein transgenic mice. Neuroscience 146:1555–1560

    Article  PubMed  CAS  Google Scholar 

  20. Ide Y, Fujiyama F, Okamoto-Furuta K, Tamamaki N, Kaneko T, Hisatsune T (2008) Rapid integration of young newborn dentate gyrus granule cells in the adult hippocampal circuitry. Eur J Neurosci 28:2381–2392

    Article  PubMed  Google Scholar 

  21. Ambrogini P, Cuppini R, Lattanzi D, Ciuffoli S, Frontini A, Fanelli M (2010) Synaptogenesis in adult-generated hippocampal granule cells is affected by behavioral experiences. Hippocampus 20:799–810

    PubMed  CAS  Google Scholar 

  22. Winner B, Melrose HL, Zhao C, Hinkle KM, Yue M, Kent C, Braithwaite AT, Ogholikhan S, Aigner R, Winkler J, Farrer MJ, Gage FH (2011) Adult neurogenesis and neurite outgrowth are impaired in LRRK2 G2019S mice. Neurobiol Dis 41:706–716

    Article  PubMed  CAS  Google Scholar 

  23. Liu Y, Namba T, Liu J, Suzuki R, Shioda S, Seki T (2010) Glial fibrillary acidic protein-expressing neural progenitors give rise to immature neurons via early intermediate progenitors expressing both glial fibrillary acidic protein and neuronal markers in the adult hippocampus. Neuroscience 166:241–251

    Article  PubMed  CAS  Google Scholar 

  24. Seki T, Arai Y (1999) Temporal and spacial relationships between PSA-NCAM-expressing, newly generated granule cells, and radial glia-like cells in the adult dentate gyrus. J Comp Neurol 410:503–513

    Article  PubMed  CAS  Google Scholar 

  25. von Bohlen Und Halbach O (2007) Immunohistological markers for staging neurogenesis in adult hippocampus. Cell Tissue Res 329:409–420

    Article  Google Scholar 

  26. Rao MS, Shetty AK (2004) Efficacy of doublecortin as a marker to analyse the absolute number and dendritic growth of newly generated neurons in the adult dentate gyrus. Eur J Neurosci 19:234–246

    Article  PubMed  Google Scholar 

  27. Gleeson JG, Lin PT, Flanagan LA, Walsh CA (1999) Doublecortin is a microtubule-associated protein and is expressed widely by migrating neurons. Neuron 23:257–271

    Article  PubMed  CAS  Google Scholar 

  28. Snyder JS, Choe JS, Clifford MA, Jeurling SI, Hurley P, Brown A, Kamhi JF, Cameron HA (2009) Adult-born hippocampal neurons are more numerous, faster maturing, and more involved in behavior in rats than in mice. J Neurosci 29:14484–14495

    Article  PubMed  CAS  Google Scholar 

  29. Nguyen L, Rigo JM, Malgrange B, Moonen G, Belachew S (2003) Untangling the functional potential of PSA-NCAM-expressing cells in CNS development and brain repair strategies. Curr Med Chem 10:2185–2196

    Article  PubMed  CAS  Google Scholar 

  30. Wang JW, David DJ, Monckton JE, Battaglia F, Hen R (2008) Chronic fluoxetine stimulates maturation and synaptic plasticity of adult-born hippocampal granule cells. J Neurosci 28:1374–1384

    Article  PubMed  CAS  Google Scholar 

  31. Revest JM, Dupret D, Koehl M, Funk-Reiter C, Grosjean N, Piazza PV, Abrous DN (2009) Adult hippocampal neurogenesis is involved in anxiety-related behaviors. Mol Psychiatry 14: 959–967

    Article  PubMed  Google Scholar 

  32. Gao X, Arlotta P, Macklis JD, Chen J (2007) Conditional knock-out of beta-catenin in postnatal-born dentate gyrus granule neurons results in dendritic malformation. J Neurosci 27:14317–14325

    Article  PubMed  CAS  Google Scholar 

  33. Groc L, Petanjek Z, Gustafsson B, Ben-Ari Y, Hanse E, Khazipov R (2002) In vivo blockade of neural activity alters dendritic development of neonatal CA1 pyramidal cells. Eur J Neurosci 16:1931–1938

    Article  PubMed  Google Scholar 

  34. Ge S, Goh EL, Sailor KA, Kitabatake Y, Ming GL, Song H (2006) GABA regulates synaptic integration of newly generated neurons in the adult brain. Nature 439:589–593

    Article  PubMed  CAS  Google Scholar 

  35. Sholl DA (1953) Dendritic organization in the neurons of the visual and motor cortices of the cat. J Anat 87:387–406

    PubMed  CAS  Google Scholar 

  36. Wojtowicz JM, Kee N (2006) BrdU assay for neurogenesis in rodents. Nat Protoc 1:1399–1405

    Article  PubMed  CAS  Google Scholar 

  37. Kee N, Sivalingam S, Boonstra R, Wojtowicz JM (2002) The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis. J Neurosci Methods 115:97–105

    Article  PubMed  CAS  Google Scholar 

  38. Jessberger S, Gage FH (2008) Stem-cell-associated structural and functional plasticity in the aging hippocampus. Psychol Aging 23:684–691

    Article  PubMed  Google Scholar 

  39. van Praag H (2008) Neurogenesis and exercise: past and future directions. Neuromolecular Med 10:128–140

    Article  PubMed  Google Scholar 

  40. Schaeffer EL, Novaes BA, da Silva ER, Skaf HD, Mendes-Neto AG (2009) Strategies to promote differentiation of newborn neurons into mature functional cells in Alzheimer brain. Prog Neuropsychopharmacol Biol Psychiatry 33:1087–1102

    Article  PubMed  CAS  Google Scholar 

  41. Rosenzweig S, Wojtowicz JM (2011) Analyzing dendritic growth in a population of immature neurons in the adult dentate gyrus using laminar quantification of disjointed dendrites. Front Neurosci 5:34

    Article  PubMed  Google Scholar 

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Rosenzweig, S., Wojtowicz, J.M. (2013). Laminar Quantification of Dendrites in Dentate Gyrus Granule Neurons. In: Nguyen, P. (eds) Multidisciplinary Tools for Investigating Synaptic Plasticity. Neuromethods, vol 81. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-517-0_6

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  • DOI: https://doi.org/10.1007/978-1-62703-517-0_6

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-516-3

  • Online ISBN: 978-1-62703-517-0

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