Cell and Tissue Research

, Volume 376, Issue 2, pp 153–163 | Cite as

Identification of calretinin-expressing retinal ganglion cells projecting to the mouse superior colliculus

  • Eun-Shil Lee
  • Jea-Young Lee
  • Gil Hyun Kim
  • Chang-Jin JeonEmail author
Regular Article


In mice, retinal ganglion cells (RGCs), which consist of around 30 subtypes, exclusively transmit retinal information to the relevant brain systems through parallel visual pathways. The superior colliculus (SC) receives the vast majority of this information from several RGC subtypes. The objective of the current study is to identify the types of calretinin (CR)-expressing RGCs that project to the SC in mice. To label RGCs, we performed CR immunoreactivity in the mouse retina after injections of fluorescent dye, dextran into mouse SC. Subsequently, the neurons double-labeled for dextran and CR were iontophoretically injected with the lipophilic dye, DiI, to characterize the detailed morphological properties of these cells. The analysis of various morphological parameters, including dendritic arborization, dendritic field size and stratification, indicated that, of the ten different types of CR-expressing RGCs in the retina, the double-labeled cells consisted of at least eight types of RGCs that projected to the SC. These cells tended to have small-medium field sizes. However, except for dendritic field size, the cells did not exhibit consistent characteristics for the other morphometric parameters examined. The combination of a tracer and single-cell injections after immunohistochemistry for a particular molecule provided valuable data that confirmed the presence of distinct subtypes of RGCs within multiple-labeled RGCs that projected to specific brain regions.


Calretinin Retinal ganglion cells Retrograde tracer injection Single-cell injection Superior colliculus 



We thank Cactus Communications for proofreading the manuscript.


This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (NRF-2016R1D1A1A09918427).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All procedures involving animals were in accordance with the ethical standards of our institution and were approved by the animal rights committee at Kyungpook National University, Deagu, South Korea (permission NO. 2015-0104). This article does not contain any studies with human participants performed by any of the authors.


  1. Ahmadlou M, Heimel JA (2015) Preference for concentric orientations in the mouse superior colliculus. Nat Commun 6:6773CrossRefGoogle Scholar
  2. Badea TC, Nathans J (2004) Quantitative analysis of neuronal morphologies in the mouse retina visualized by using a genetically directed reporter. J Comp Neurol 480:331–351CrossRefGoogle Scholar
  3. Baden T, Berens P, Franke K, Román Rosón M, Bethge M, Euler T (2016) The functional diversity of retinal ganglion cells in the mouse. Nature 529:345–350CrossRefGoogle Scholar
  4. Berson DM (2008) Retinal ganglion cell types and their central projections. In: Basbaum AI, Kaneko A, Shepherd GM, Westheimer G (eds) The senses: a comprehensive reference, Vision 1, vol 1. Academic Press, San Diego, pp 491–520CrossRefGoogle Scholar
  5. Boycott BB, Wässle H (1974) The morphological types of ganglion cells of the domestic cat's retina. J Physiol 240:397–419CrossRefGoogle Scholar
  6. Callaway EM (2005) Structure and function of parallel pathways in the primate early visual system. J Physiol 566:13–19CrossRefGoogle Scholar
  7. Cang J, Feldheim DA (2013) Developmental mechanisms of topographic map formation and alignment. Annu Rev Neurosci 36:51–77CrossRefGoogle Scholar
  8. Chen SK, Badea TC, Hattar S (2011) Photoentrainment and pupillary light reflex are mediated by distinct populations of ipRGCs. Nature 476:92–95CrossRefGoogle Scholar
  9. Cheron G, Gall D, Servais L, Dan B, Maex R, Schiffmann SN (2004) Inactivation of calcium-binding protein genes induces 160 Hz oscillations in the cerebellar cortex of alert mice. J Neurosci 24:434–441CrossRefGoogle Scholar
  10. Coombs J, van der List D, Wang GY, Chalupa LM (2006) Morphological properties of mouse retinal ganglion cells. Neuroscience 140:123–136CrossRefGoogle Scholar
  11. Dhande OS, Huberman AD (2014) Retinal ganglion cell maps in the brain: implications for visual processing. Curr Opin Neurobiol 24:133–142CrossRefGoogle Scholar
  12. Ecker JL, Dumitrescu ON, Wong KY, Alam NM, Chen SK, LeGates T, Renna JM, Prusky GT, Berson DM, Hattar S (2010) Melanopsin-expressing retinal ganglion-cell photoreceptors: cellular diversity and role in pattern vision. Neuron 67:49–60CrossRefGoogle Scholar
  13. Ellis EM, Gauvain G, Sivyer B, Murphy GJ (2016) Shared and distinct retinal input to the mouse superior colliculus and dorsal lateral geniculate nucleus. J Neurophysiol 116:602–610CrossRefGoogle Scholar
  14. Farrow K, Masland RH (2011) Physiological clustering of visual channels in the mouse retina. J Neurophysiol 105:1516–1530CrossRefGoogle Scholar
  15. Farrow K, Teixeira M, Szikra T, Viney TJ, Balint K, Yonehara K et al (2013) Ambient illumination toggles a neuronal circuit switch in the retina and visual perception at cone threshold. Neuron 78:325–338CrossRefGoogle Scholar
  16. Gall D, Roussel C, Susa I, D'Angelo E, Rossi P, Bearzatto B, Galas MC, Blum D, Schurmans S, Schiffmann SN (2003) Altered neuronal excitability in cerebellar granule cells of mice lacking calretinin. J Neurosci 23:9320–9327CrossRefGoogle Scholar
  17. Hattar S, Kumar M, Park A, Tong P, Tung J, Yau KW, Berson DM (2006) Central projections of melanopsin-expressing retinal ganglion cells in the mouse. J Comp Neurol 497:326–349CrossRefGoogle Scholar
  18. Hattar S, Liao HW, Takao M, Berson DM, Yau KW (2002) Melanopsin-containing retinal ganglion cells: architecture, projections, and intrinsic photosensitivity. Science 295:1065–1070CrossRefGoogle Scholar
  19. Haverkamp S, Wässle H (2000) Immunocytochemical analysis of the mouse retina. J Comp Neurol 424:1–23CrossRefGoogle Scholar
  20. Hof PR, Young WG, Bloom F (2000) Comparative cytoarchitectonic atlas of the C57BL/6 and 129/SV: mouse brains. Elsevier Science, New YorkGoogle Scholar
  21. Hofbauer A, Dräger UC (1985) Depth segregation of retinal ganglion cells projecting to mouse superior colliculus. J Comp Neurol 234:465–474CrossRefGoogle Scholar
  22. Huberman AD, Manu M, Koch SM, Susman MW, Lutz AB, Ullian EM, Baccus SA, Barres BA (2008) Architectures and activity-mediated refinement of axonal projections from a mosaic of genetically identified retinal ganglion cells. Neuron 59:425–438CrossRefGoogle Scholar
  23. Huberman AD, Wei W, Elstrott J, Stafford BK, Feller MB, Barres BA (2009) Genetic identification of an on-off direction-selective retinal ganglion cell subtype reveals layer-specific subcortical map of posterior motion. Neuron 62:327–334CrossRefGoogle Scholar
  24. Ito S, Feldheim DA (2018) The mouse superior colliculus: an emerging model for studying circuit formation and function. Front Neural Circuits 12:10CrossRefGoogle Scholar
  25. Jeon CJ, Strettoi E, Masland RH (1998) The major cell populations of the mouse retina. J Neurosci 18:8936–8946CrossRefGoogle Scholar
  26. Jonathan W, Hiroshi H (2017) Visual system architecture. In: Pablo A (ed) Handbook of visual optics, volume one: fundamentals and eye optics. Fundamentals. CRC Press, Boca Raton, pp 159–180Google Scholar
  27. Kao YH, Sterling P (2003) Matching neural morphology to molecular expression: single cell injection following immunostaining. J Neurocytol 32:245–251CrossRefGoogle Scholar
  28. Kay JN, De la Huerta I, Kim IJ, Zhnag Y, Yamagata M, Chu MW, Meister M, Sanes JR (2011) Retinal ganglion cells with distinct directional preferences differ in molecular identity, structure, and central projections. J Neurosci 31:7753–7762CrossRefGoogle Scholar
  29. Kim IJ, Zhang Y, Meister M, Sanes JR (2010) Laminar restriction of retinal ganglion cell dendrites and axons: subtype-specific developmental patterns revealed by transgenic markers. J Neurosci 30:1452–1462CrossRefGoogle Scholar
  30. Kim IJ, Zhang Y, Yamagata M, Meister M, Sanes JR (2008) Molecular identification of a retinal cell type that responds to upward motion. Nature 452:478–482CrossRefGoogle Scholar
  31. Kim T, Soto F, Kerschensteiner D (2015) An excitatory amacrine cell detects object motion and provides feature-selective input to ganglion cells in the mouse retina. elife.
  32. Kim TJ, Jeon CJ (2006) Morphological classification of parvalbumin-containing retinal ganglion cells in mouse: single-cell injection after immunocytochemistry. Invest Ophthalmol Vis Sci 47:2757–2764CrossRefGoogle Scholar
  33. Kong JH, Fish DR, Rockhill RL, Masland RH (2005) Diversity of ganglion cells in the mouse retina: unsupervised morphological classification and its limits. J Comp Neurol 489:293–310CrossRefGoogle Scholar
  34. Krieger B, Qiao M, Rousso DL, Sanes JR, Meister M (2017) Four alpha ganglion cell types in mouse retina: function, structure, and molecular signatures. PLoS One 12(7):e0180091CrossRefGoogle Scholar
  35. Kwon OJ, Lee ES, Jeon CJ (2014) Density and types of calretinin-containing retinal ganglion cells in rabbit. Neuroscience 278:343–353CrossRefGoogle Scholar
  36. Lee ES, Lee JY, Jeon CJ (2010) Types and density of calretinin-containing retinal ganglion cells in mouse. Neurosci Res 66:141–150CrossRefGoogle Scholar
  37. Martersteck EM, Hirokawa KE, Evarts M, Bernard A, Duan X, Li Y, Ng L, Oh SW, Ouellette B, Royall JJ, Stoecklin M, Wang Q, Zeng H, Sanes JR, Harris JA (2017) Diverse central projection patterns of retinal ganglion cells. Cell Rep 18:2058–2072CrossRefGoogle Scholar
  38. May PJ (2006) The mammalian superior colliculus: laminar structure and connections. Prog Brain Res 151:321–378CrossRefGoogle Scholar
  39. Münch TA, da Silveira RA, Siegert S, Viney TJ, Awatramani GB, Roska B (2009) Approach sensitivity in the retina processed by a multifunctional neural circuit. Nat Neurosci 12:1308–1316CrossRefGoogle Scholar
  40. Nath A, Schwartz GW (2016) Cardinal orientation selectivity is represented by two distinct ganglion cell types in mouse retina. J Neurosci 36:3208–3221CrossRefGoogle Scholar
  41. Rivlin-Etzion M, Zhou K, Wei W, Elstrott J, Nguyen PL, Barres BA, Huberman AD, Feller MG (2011) Transgenic mice reveal unexpected diversity of on-off direction-selective retinal ganglion cell subtypes and brain structures in motion processing. J Neurosci 31:8760–8769CrossRefGoogle Scholar
  42. Rockhill RL, Euler T, Masland RH (2000) Spatial order within but not between types of retinal neurons. Proc Natl Acad Sci U S A 97:2303–2307CrossRefGoogle Scholar
  43. Rodieck RW (1998) The first steps in seeing. Sinauer Associates, SunderlandGoogle Scholar
  44. Roska B, Meister M (2014) The retina dissects the visual scene in distinct features. In: Werner JS, Chalupa LM (eds) The new visual neuroscience, retinal mechanisms and processes. MIT Press, Cambridge, pp 163–182Google Scholar
  45. Sanes JR, Masland RH (2015) The types of retinal ganglion cells: current status and implications for neuronal classification. Annu Rev Neurosci 38:221–246CrossRefGoogle Scholar
  46. Schiffmann SN, Cheron G, Lohof A, d’Alcantara P, Meyer M, Parmentier M, Schurmans S (1999) Impaired motor coordination and Purkinje cell excitability in mice lacking calretinin. Proc Natl Acad Sci U S A 96:5257–5262CrossRefGoogle Scholar
  47. Schmolesky M (1995-2005) The primary visual cortex. In: Kolb H, Fernandez E, Nelson R (eds) Webvision, The Organization of the Retina and Visual System (Internet). University of Utah Health Sciences Center, Salt Lake CityGoogle Scholar
  48. Schwaller B (2014) Calretinin: from a “simple” Ca(2+) buffer to a multifunctional protein implicated in many biological processes. Front Neuroanat. eCollection 2014.
  49. Stein BE, Wallace MW, Stanford TR, Jiang W (2002) Cortex governs multisensory integration in the midbrain. Neuroscientist 8:306–314CrossRefGoogle Scholar
  50. Sun W, Li N, He S (2002) Large-scale morphological survey of rat retinal ganglion cells. Vis Neurosci 19:483–493CrossRefGoogle Scholar
  51. Trenholm S, Johnson K, Li X, Smith RG, Awatramani GB (2011) Parallel mechanisms encode direction in the retina. Neuron 71:683–694CrossRefGoogle Scholar
  52. Völgyi B, Chheda S, Bloomfield SA (2009) Tracer coupling patterns of the ganglion cell subtypes in the mouse retina. J Comp Neurol 512:664–687CrossRefGoogle Scholar
  53. Wang L, Sarnaik R, Rangarajan K, Liu X, Cang J (2010) Visual receptive field properties of neurons in the superficial superior colliculus of the mouse. J Neurosci 30:16573–16584CrossRefGoogle Scholar
  54. Yang G, Masland RH (1994) Receptive fields and dendritic structure of directionally selective retinal ganglion cells. J Neurosci 14:5267–5280CrossRefGoogle Scholar
  55. Yi CW, Yu SH, Lee ES, Lee JG, Jeon CJ (2012) Types of parvalbumin-containing retinotectal ganglion cells in mouse. Acta Histochem Cytochem 45:201–210CrossRefGoogle Scholar
  56. Zhang Y, Kim IJ, Sanes JR, Meister M (2012) The most numerous ganglion cell type of the mouse retina is a selective feature detector. Proc Natl Acad Sci U S A 109:E2391–E2398CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Eun-Shil Lee
    • 1
  • Jea-Young Lee
    • 2
  • Gil Hyun Kim
    • 1
  • Chang-Jin Jeon
    • 1
    Email author
  1. 1.Department of Biology, School of Life Sciences, BK 21 Plus KNU Creative BioResearch Group, College of Natural Sciences, and Brain Science and Engineering InstituteKyungpook National UniversityDaeguSouth Korea
  2. 2.Center of Excellence for Aging and Brain Repair, USF HealthUniversity of South FloridaTampaUSA

Personalised recommendations