In the mammalian retina, light signals generated in photoreceptors are passed to bipolar and horizontal cells via synaptic contacts. In various pathological conditions, these second-order neurons extend neurites into the outer nuclear layer (ONL). However, the molecular events associated with this neurite outgrowth are not known. Here, we characterized the morphological synaptic changes in the CNGA3/CNGB1 double-knockout (A3B1) mouse, a model of retinitis pigmentosa. In these mice, horizontal cells looked normal until postnatal day (p) 11, but started growing neurites into the ONL 1 day later. At p28, the number of sprouting processes decreased, but the remaining sprouts developed synapse-like contacts at rod cell bodies, with an ultrastructural appearance reminiscent of ribbon synapses. Hence, neurite outgrowth and ectopic synaptogenesis in the A3B1 retina were precisely timed events starting at p12 and p28, respectively. We therefore performed microarray analysis of retinal gene expression in A3B1 and wild-type mice at those ages to evaluate the genomic response underlying these two events. This analysis identified 163 differentially regulated genes in the A3B1 retina related to neurite outgrowth or plasticity of synapses. The global changes in gene expression in the A3B1 retina were consistent with activation of signaling pathways related to Tp53, Smad, and Stat3. Moreover, key molecules of these signaling pathways could be localized at or in close proximity to outgrowing neurites. We therefore propose that Tp53, Smad, and Stat3 signaling pathways contribute to the synaptic plasticity in the A3B1 retina.
This is a preview of subscription content, log in to check access.
Buy single article
Instant unlimited access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Wässle H (2004) Parallel processing in the mammalian retina. Nat Rev Neurosci 5(10):747–757
Blanks JC, Adinolfi AM, Lolley RN (1974) Synaptogenesis in the photoreceptor terminal of the mouse retina. J Comp Neurol 156(1):81–93
Olney JW (1968) An electron microscopic study of synapse formation, receptor outer segment development, and other aspects of developing mouse retina. Invest Ophthalmol 7(3):250–268
Claes E, Seeliger M, Michalakis S, Biel M, Humphries P, Haverkamp S (2004) Morphological characterization of the retina of the CNGA3(-/-)Rho(-/-) mutant mouse lacking functional cones and rods. Invest Ophthalmol Vis Sci 45(6):2039–2048
Lewis GP, Linberg KA, Fisher SK (1998) Neurite outgrowth from bipolar and horizontal cells after experimental retinal detachment. Invest Ophthalmol Vis Sci 39(2):424–434
Liets LC, Eliasieh K, van der List DA, Chalupa LM (2006) Dendrites of rod bipolar cells sprout in normal aging retina. Proc Natl Acad Sci USA 103(32):12156–12160
Hüttl S, Michalakis S, Seeliger M, Luo DG, Acar N, Geiger H, Hudl K, Mader R, Haverkamp S, Moser M, Pfeifer A, Gerstner A, Yau KW, Biel M (2005) Impaired channel targeting and retinal degeneration in mice lacking the cyclic nucleotide-gated channel subunit CNGB1. J Neurosci 25(1):130–138
Dick O, Dieck S, Altrock WD, Ammermuller J, Weiler R, Garner CC, Gundelfinger ED, Brandstatter JH (2003) The presynaptic active zone protein bassoon is essential for photoreceptor ribbon synapse formation in the retina. Neuron 37(5):775–786
Ball SL, Pardue MT, McCall MA, Gregg RG, Peachey NS (2003) Immunohistochemical analysis of the outer plexiform layer in the nob mouse shows no abnormalities. Vis Neurosci 20(3):267–272
Bayley PR, Morgans CW (2007) Rod bipolar cells and horizontal cells form displaced synaptic contacts with rods in the outer nuclear layer of the nob2 retina. J Comp Neurol 500(2):286–298
Chang B, Heckenlively JR, Bayley PR, Brecha NC, Davisson MT, Hawes NL, Hirano AA, Hurd RE, Ikeda A, Johnson BA, McCall MA, Morgans CW, Nusinowitz S, Peachey NS, Rice DS, Vessey KA, Gregg RG (2006) The nob2 mouse, a null mutation in Cacna1f: anatomical and functional abnormalities in the outer retina and their consequences on ganglion cell visual responses. Vis Neurosci 23(1):11–24
Haeseleer F, Imanishi Y, Maeda T, Possin DE, Maeda A, Lee A, Rieke F, Palczewski K (2004) Essential role of Ca2+-binding protein 4, a Cav1.4 channel regulator, in photoreceptor synaptic function. Nat Neurosci 7(10):1079–1087
Mansergh F, Orton NC, Vessey JP, Lalonde MR, Stell WK, Tremblay F, Barnes S, Rancourt DE, Bech-Hansen NT (2005) Mutation of the calcium channel gene Cacna1f disrupts calcium signaling, synaptic transmission and cellular organization in mouse retina. Hum Mol Genet 14(20):3035–3046
Biel M, Seeliger M, Pfeifer A, Kohler K, Gerstner A, Ludwig A, Jaissle G, Fauser S, Zrenner E, Hofmann F (1999) Selective loss of cone function in mice lacking the cyclic nucleotide-gated channel CNG3. Proc Natl Acad Sci USA 96(13):7553–7557
Haverkamp S, Wassle H (2000) Immunocytochemical analysis of the mouse retina. J Comp Neurol 424(1):1–23
Dieck S, Altrock WD, Kessels MM, Qualmann B, Regus H, Brauner D, Fejtova A, Bracko O, Gundelfinger ED, Brandstatter JH (2005) Molecular dissection of the photoreceptor ribbon synapse: physical interaction of Bassoon and RIBEYE is essential for the assembly of the ribbon complex. J Cell Biol 168(5):825–836
Walzer T, Galibert L, Comeau MR, De Smedt T (2005) Plexin C1 engagement on mouse dendritic cells by viral semaphorin A39R induces actin cytoskeleton rearrangement and inhibits integrin-mediated adhesion and chemokine-induced migration. J Immunol 174(1):51–59
Zinchuk V, Zinchuk O, Okada T (2007) Quantitative colocalization analysis of multicolor confocal immunofluorescence microscopy images: pushing pixels to explore biological phenomena. Acta histochemica et cytochemica 40(4):101–111
Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30(9):e36
Vardi N, Morigiwa K (1997) ON cone bipolar cells in rat express the metabotropic receptor mGluR6. Vis Neurosci 14(4):789–794
Michalakis S, Geiger H, Haverkamp S, Hofmann F, Gerstner A, Biel M (2005) Impaired opsin targeting and cone photoreceptor migration in the retina of mice lacking the cyclic nucleotide-gated channel CNGA3. Invest Ophthalmol Vis Sci 46(4):1516–1524
Di Giovanni S, Rathore K (2012) p53-dependent pathways in neurite outgrowth and axonal regeneration. Cell Tissue Res 349(1):87–89
Floriddia EM, Rathore KI, Tedeschi A, Quadrato G, Wuttke A, Lueckmann JM, Kigerl KA, Popovich PG, Di Giovanni S (2012) p53 regulates the neuronal intrinsic and extrinsic responses affecting the recovery of motor function following spinal cord injury. J Neurosci 32(40):13956–13970
He JC, Gomes I, Nguyen T, Jayaram G, Ram PT, Devi LA, Iyengar R (2005) The G alpha(o/i)-coupled cannabinoid receptor-mediated neurite outgrowth involves Rap regulation of Src and Stat3. J Biol Chem 280(39):33426–33434
Yang LT, Alexandropoulos K, Sap J (2002) c-SRC mediates neurite outgrowth through recruitment of Crk to the scaffolding protein Sin/Efs without altering the kinetics of ERK activation. J Biol Chem 277(20):17406–17414
Whitman M (1998) Smads and early developmental signaling by the TGFbeta superfamily. Genes Dev 12(16):2445–2462
Mackinnon AC, Gibbons MA, Farnworth SL, Leffler H, Nilsson UJ, Delaine T, Simpson AJ, Forbes SJ, Hirani N, Gauldie J, Sethi T (2012) Regulation of TGF-beta1 driven lung fibrosis by galectin-3. Am J Respir Crit Care Med 185(5):537–546
Kim J, Moon C, Ahn M, Joo HG, Jin JK, Shin T (2009) Immunohistochemical localization of galectin-3 in the pig retina during postnatal development. Mol Vis 15:1971–1976
Uehara F, Ohba N, Ozawa M (2001) Isolation and characterization of galectins in the mammalian retina. Invest Ophthalmol Vis Sci 42(10):2164–2172
Klein R (2009) Bidirectional modulation of synaptic functions by Eph/ephrin signaling. Nat Neurosci 12(1):15–20
Stumm R, Hollt V (2007) CXC chemokine receptor 4 regulates neuronal migration and axonal pathfinding in the developing nervous system: implications for neuronal regeneration in the adult brain. J Mol Endocrinol 38(3):377–382
Engle EC (2010) Human genetic disorders of axon guidance. Cold Spring Harbor Perspect Biol 2(3):a001784
Schmitt A, Hasan A, Gruber O, Falkai P (2011) Schizophrenia as a disorder of disconnectivity. Eur Arch Psychiatry Clin Neurosci 261(Suppl 2):S150–S154
Haverkamp S, Michalakis S, Claes E, Seeliger MW, Humphries P, Biel M, Feigenspan A (2006) Synaptic plasticity in CNGA3(−/−) mice: cone bipolar cells react on the missing cone input and form ectopic synapses with rods. J Neurosci 26(19):5248–5255
We thank G.-S. Nam, Ilse von Graevenitz and Jennifer Schmidt for technical assistance, Jeroen Pasterkamp (University Medical Center Utrecht) for helpful discussion on PlexinC1, and Heinz Wässle for financial support. Grant sponsor: Deutsche Forschungsgemeinschaft (DFG).
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Michalakis, S., Schäferhoff, K., Spiwoks-Becker, I. et al. Characterization of neurite outgrowth and ectopic synaptogenesis in response to photoreceptor dysfunction. Cell. Mol. Life Sci. 70, 1831–1847 (2013). https://doi.org/10.1007/s00018-012-1230-z
- Cyclic nucleotide-gated channel
- Retinitis pigmentosa
- Horizontal cell
- Bipolar cell
- Synaptic plasticity
- Ectopic synapse