Differential expression of PKCα and -β in the zebrafish retina

  • Marion F. Haug
  • Manuela Berger
  • Matthias Gesemann
  • Stephan C. F. NeuhaussEmail author
Short Communication


The retina is a complex neural circuit, which processes and transmits visual information from light perceiving photoreceptors to projecting retinal ganglion cells. Much of the computational power of the retina rests on signal integrating interneurons, such as bipolar cells. Commercially available antibodies against bovine and human conventional protein kinase C (PKC) α and -β are frequently used as markers for retinal ON-bipolar cells in different species, despite the fact that it is not known which bipolar cell subtype(s) they actually label. In zebrafish (Danio rerio) five prkc genes (coding for PKC proteins) have been identified. Their expression has not been systematically determined. While prkcg is not expressed in retinal tissue, the other four prkc (prkcaa, prkcab, prkcba, prkcbb) transcripts were found in different parts of the inner nuclear layer and some as well in the retinal ganglion cell layer. Immunohistochemical analysis in adult zebrafish retina using fluorescent in situ hybridization and PKC antibodies showed an overlapping immunolabeling of ON-bipolar cells that are most likely of the BON s6 and BON s6L or RRod type. However, comparison of transcript expression with immunolabeling, implies that these antibodies are not specific for one single zebrafish conventional PKC, but rather detect a combination of PKC -α and -β variants.


Zebrafish (RRID:ZIRC_ZL84) Protein kinase C Bipolar cell Retina 



We would like to acknowledge Kara Kristiansen and Martin Walther for technical support and excellent fish care.

Author contributions

MFH carried out the experiments, participated in the design of the study and drafted and edited the manuscript; MB helped with the in situ hybridization and the fluorescent in situ hybridization, and drafted the manuscript; MG participated in the design of the study and edited the manuscript; SCFN participated in the design of the study, coordinated the study and edited the manuscript. All authors gave final approval for publication.

Compliance with ethical standards

Conflict of interest

The author declares that they have no conflict of interest.


  1. Bilotta J, Saszik S, Sutherland SE (2001) Rod contributions to the electroretinogram of the dark-adapted developing zebrafish. Dev Dyn 222(4):564–570PubMedGoogle Scholar
  2. Boycott BB, Wässle H (1991) Morphological classification of bipolar cells of the primate retina. Eur J Neurosci 3(11):1069–1088PubMedGoogle Scholar
  3. Breuiller-Fouché M, Tertrin-Clary C, Héluy V, Fournier T, Ferré F (1998) Role of protein kinase C in endothelin-1-induced contraction of human myometrium. Biol Reprod 59(1):153–159PubMedGoogle Scholar
  4. Caminos E, Velasco A, Jarrín M, Aijón J, Lara JM (1999) Protein kinase C-like immunoreactive cells in embryo and adult chicken retinas. Brain Res Dev Brain Res 118(1–2):227–230PubMedGoogle Scholar
  5. Caminos E, Velasco A, Jarrín M, Lillo C, Jimeno D, Aijón J, Lara JM (2000) A comparative study of protein kinase C-like immunoreactive cells in the retina. Brain Behav Evol 56(6):330–339PubMedGoogle Scholar
  6. Connaughton VP (2011) Bipolar cells in the zebrafish retina. Vis Neurosci 28(1):77–93PubMedGoogle Scholar
  7. Connaughton VP, Graham D, Nelson R (2004) Identification and morphological classification of horizontal, bipolar, and amacrine cells within the zebrafish retina. J Comp Neurol 477(4):371–385PubMedGoogle Scholar
  8. Euler T, Wässle H (1995) Immunocytochemical identification of cone bipolar cells in the rat retina. J Comp Neurol 361(3):461–478PubMedGoogle Scholar
  9. Euler T, Schneider H, Wässle H (1996) Glutamate responses of bipolar cells in a slice preparation of the rat retina. J Neurosci 16(9):2934–2944PubMedGoogle Scholar
  10. Euler T, Haverkamp S, Schubert T, Baden T (2014) Retinal bipolar cells: elementary building blocks of vision. Nat Rev Neurosci 15(8):507–519PubMedGoogle Scholar
  11. Famiglietti EV (1981) Functional architecture of cone bipolar cells in mammalian retina. Vis Res 21(11):1559–1563PubMedGoogle Scholar
  12. Fleisch VC, Schonthaler HB, Lintig J von, Stephan SCF (2008) Subfunctionalization of a retinoid-binding protein provides evidence for two parallel visual cycles in the cone-dominant zebrafish retina. J Neurosci 28(33):8208–8216PubMedGoogle Scholar
  13. Force A, Lynch M, Pickett FB, Amores A, Yan YL, Postlethwait J (1999) Preservation of duplicate genes by complementary, degenerative mutations. Genetics 151(4):1531–1545PubMedPubMedCentralGoogle Scholar
  14. Fukuda K, Saito N, Yamamoto M, Tanaka C (1994) Immunocytochemical localization of the alpha-, beta I-, beta II- and gamma-subspecies of protein kinase C in the monkey visual pathway. Brain Res 658(1–2):155–162PubMedGoogle Scholar
  15. Ghosh KK, Bujan S, Haverkamp S, Feigenspan A, Wässle H (2004) Types of bipolar cells in the mouse retina. J Comp Neurol 469(1):70–82PubMedGoogle Scholar
  16. Glasauer SMK, Neuhauss SCF (2014) Whole-genome duplication in teleost fishes and its evolutionary consequences. Mol Genet Genom 289(6):1045–1060Google Scholar
  17. Greene NM, Williams DS, Newton AC (1997) Identification of protein kinase C phosphorylation sites on bovine rhodopsin. J Biol Chem 272(16):10341–10344PubMedGoogle Scholar
  18. Greferath U, Grünert U, Wässle H (1990) Rod bipolar cells in the mammalian retina show protein kinase C-like immunoreactivity. J Comp Neurol 301(3):433–442PubMedGoogle Scholar
  19. Grünert U, Martin PR, Wässle H (1994) Immunocytochemical analysis of bipolar cells in the macaque monkey retina. J Comp Neurol 348(4):607–627PubMedGoogle Scholar
  20. Haug MF, Gesemann M, Mueller T, Stephan SCF (2013) Phylogeny and expression divergence of metabotropic glutamate receptor genes in the brain of zebrafish (Danio rerio). J Comp Neurol 521(7):1533–1560PubMedGoogle Scholar
  21. Haug MF, Gesemann M, Berger M, Neuhauss SCF (2018) Phylogeny and distribution of protein kinase C variants in the zebrafish. J Comp Neurol 526(7):1097–1109PubMedGoogle Scholar
  22. Haverkamp S, Wässle H (2000) Immunocytochemical analysis of the mouse retina. J Comp Neurol 424(1):1–23PubMedGoogle Scholar
  23. Hodel C, Niklaus S, Heidemann M, Klooster J, Kamermans M, Biehlmaier O, Gesemann M, Neuhauss SCF (2014) Myosin VIIA is a marker for the cone accessory outer segment in zebrafish. Anat Rec 297(9):1777–1784Google Scholar
  24. Huang Y-Y, Haug MF, Gesemann M, Stephan SCF (2012) Novel expression patterns of metabotropic glutamate receptor 6 in the zebrafish nervous system. PLoS One 7(4):e35256PubMedPubMedCentralGoogle Scholar
  25. Jordan M, Schallhorn A, Wurm FM (1996) Transfecting mammalian cells: optimization of critical parameters affecting calcium-phosphate precipitate formation. Nucleic Acids Res 24(4):596–601PubMedPubMedCentralGoogle Scholar
  26. Kang G-H, Srivastava A, Kim YE, Park H-J, Park CK, Sohn TS, Kim S, Kang DY, Kim K-M (2011) DOG1 and PKC-θ are useful in the diagnosis of KIT-negative gastrointestinal stromal tumors. Mod Pathol 24(6):866–875PubMedGoogle Scholar
  27. Kapoor CL, Chader GJ (1984) Endogenous phosphorylation of retinal photoreceptor outer segment proteins by calcium phospholipid-dependent protein kinase. Biochem Biophys Res Commun 122(3):1397–1403PubMedGoogle Scholar
  28. Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203(3):253–310PubMedPubMedCentralGoogle Scholar
  29. Kolb H, Zhang L (1997) Immunostaining with antibodies against protein kinase C isoforms in the fovea of the monkey retina. Microsc Res Tech 36(1):57–75PubMedGoogle Scholar
  30. Kolb H, Linberg KA, Fisher SK (1992) Neurons of the human retina: a Golgi study. J Comp Neurol 318(2):147–187PubMedGoogle Scholar
  31. Kolb H, Zhang L, Dekorver L (1993) Differential staining of neurons in the human retina with antibodies to protein kinase C isozymes. Vis Neurosci 10(2):341–351PubMedGoogle Scholar
  32. Kosaka J, Suzuki A, Morii E, Nomura S (1998) Differential localization and expression of alpha and beta isoenzymes of protein kinase C in the rat retina. J Neurosci Res 54(5):655–663PubMedGoogle Scholar
  33. Li YN, Tsujimura T, Kawamura S, Dowling JE (2012) Bipolar cell–photoreceptor connectivity in the zebrafish (Danio rerio) retina. J Comp Neurol 520(16):3786–3802PubMedPubMedCentralGoogle Scholar
  34. Mack AF (2007) Evidence for a columnar organization of cones, Müller cells, and neurons in the retina of a cichlid fish. Neuroscience 144(3):1004–1014PubMedGoogle Scholar
  35. Macosko EZ, Basu A, Satija R, Nemesh J, Shekhar K, Goldman M, Tirosh I, Bialas AR, Kamitaki N, Martersteck EM, Trombetta JJ, Weitz DA, Sanes JR, Shalek AK, Regev A, McCarroll SA (2015) Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell 161(5):1202–1214PubMedPubMedCentralGoogle Scholar
  36. Negishi K, Kato S, Teranishi T (1988) Dopamine cells and rod bipolar cells contain protein kinase C-like immunoreactivity in some vertebrate retinas. Neurosci Lett 94(3):247–252PubMedGoogle Scholar
  37. Nelson R, Kolb H (1983) Synaptic patterns and response properties of bipolar and ganglion cells in the cat retina. Vis Res 23(10):1183–1195PubMedGoogle Scholar
  38. Newton AC (2010) Protein kinase C: poised to signal. American journal of physiology. Endocrinol Metab 298(3):402Google Scholar
  39. Ohki K, Yoshida K, Imaki J, Harada T, Matsuda H (1994) The existence of protein kinase C in cone photoreceptors in the rat retina. Curr Eye Res 13(7):547–550PubMedGoogle Scholar
  40. Osborne NN, Broyden NJ, Barnett NL, Morris NJ (1991) Protein kinase C (alpha and beta) immunoreactivity in rabbit and rat retina: effect of phorbol esters and transmitter agonists on immunoreactivity and the translocation of the enzyme from cytosolic to membrane compartments. J Neurochem 57(2):594–604PubMedGoogle Scholar
  41. Osborne NN, Barnett NL, Morris NJ, Huang FL (1992) The occurrence of three isoenzymes of protein kinase C (alpha, beta and gamma) in retinas of different species. Brain Res 570(1–2):161–166PubMedGoogle Scholar
  42. Pang J-J, Gao F, Wu SM (2004) Stratum-by-stratum projection of light response attributes by retinal bipolar cells of Ambystoma. J Physiol 558(Pt 1):249–262PubMedPubMedCentralGoogle Scholar
  43. Pinzon-Guzman C, Zhang SS-M, Barnstable CJ (2011) Specific protein kinase C isoforms are required for rod photoreceptor differentiation. J Neurosci 31(50):18606–18617PubMedPubMedCentralGoogle Scholar
  44. Rodrigues PdS, Dowling JE (1990) Dopamine induces neurite retraction in retinal horizontal cells via diacylglycerol and protein kinase C. Proc Natl Acad Sci USA 87(24):9693–9697Google Scholar
  45. Saito T, Kujiraoka T, Yonaha T, Chino Y (1985) Reexamination of photoreceptor-bipolar connectivity patterns in carp retina: HRP-EM and Golgi-EM studies. J Comp Neurol 236(2):141–160PubMedGoogle Scholar
  46. Sakai N, Saito N, Seki T (2011) Molecular pathophysiology of neurodegenerative disease caused by γPKC mutations. World J Biol Psychiatry 12(Suppl 1):95–98PubMedGoogle Scholar
  47. Schmidt KF (1996) Protein kinase C does not mediate the dopamine-dependent modulation of glutamate receptors in retinal horizontal cells of the perch (Perca fluviatilis). Vis Res 36(24):3939–3942PubMedGoogle Scholar
  48. Steinberg SF (2008) Structural basis of protein kinase C isoform function. Physiol Revi 88(4):1341–1378Google Scholar
  49. Suzuki S, Kaneko A (1990) Identification of bipolar cell subtypes by protein kinase C-like immunoreactivity in the goldfish retina. Vis Neurosci 5(3):223–230PubMedGoogle Scholar
  50. Tarr JM, Kaul K, Chopra M, Kohner EM, Chibber R (2013) Pathophysiology of diabetic retinopathy. ISRN Ophthalmol 2013:343560PubMedPubMedCentralGoogle Scholar
  51. Udovichenko IP, Cunnick J, Gonzales K, Takemoto DJ (1993) Phosphorylation of bovine rod photoreceptor cyclic GMP phosphodiesterase. Biochem J 295(Pt 1):49–55PubMedPubMedCentralGoogle Scholar
  52. Udovichenko IP, Cunnick J, Gonzalez K, Yakhnin A, Takemoto DJ (1996) Protein kinase C in rod outer segments: effects of phosphorylation of the phosphodiesterase inhibitory subunit. Biochem J 317(Pt 1):291–295PubMedPubMedCentralGoogle Scholar
  53. Udovichenko IP, Newton AC, Williams DS (1997) Contribution of protein kinase C to the phosphorylation of rhodopsin in intact retinas. J Biol Chem 272(12):7952–7959PubMedGoogle Scholar
  54. Usuda N, Kong Y, Hagiwara M, Uchida C, Terasawa M, Nagata T, Hidaka H (1991) Differential localization of protein kinase C isozymes in retinal neurons. J Cell Biol 112(6):1241–1247PubMedGoogle Scholar
  55. Vaquero CF, Velasco A, de la Villa P (1996) Protein kinase C localization in the synaptic terminal of rod bipolar cells. Neuroreport 7(13):2176–2180PubMedGoogle Scholar
  56. Wang L, Lam JS-Y, Zhao H, Wang J, Chan S-O (2014) Localization of protein kinase C isoforms in the optic pathway of mouse embryos and their role in axon routing at the optic chiasm. Brain Res 1575:22–32PubMedGoogle Scholar
  57. Wässle H, Puller C, Müller F, Haverkamp S (2009) Cone contacts, mosaics, and territories of bipolar cells in the mouse retina. J Neurosci 29(1):106–117PubMedGoogle Scholar
  58. Weiler R, Kohler K, Janssen U (1991) Protein kinase C mediates transient spinule-type neurite outgrowth in the retina during light adaptation. Proc Natl Acad Sci USA 88(9):3603–3607PubMedGoogle Scholar
  59. Williams DS, Liu X, Schlamp CL, Ondek B, Jaken S, Newton AC (1997) Characterization of protein kinase C in photoreceptor outer segments. J Neurochem 69(4):1693–1702PubMedGoogle Scholar
  60. Wong KY, Dowling JE (2005) Retinal bipolar cell input mechanisms in giant danio. III. ON-OFF bipolar cells and their color-opponent mechanisms. J Neurophysiol 94(1):265–272PubMedGoogle Scholar
  61. Wong KY, Cohen ED, Dowling JE (2005) Retinal bipolar cell input mechanisms in giant danio. II. Patch-clamp analysis of on bipolar cells. J Neurophysiol 93(1):94–107PubMedGoogle Scholar
  62. Wood JP, McCord RJ, Osborne NN (1997) Retinal protein kinase C. Neurochem Int 30(2):119–136PubMedGoogle Scholar
  63. Wu SM, Gao F, Maple BR (2000) Functional architecture of synapses in the inner retina: segregation of visual signals by stratification of bipolar cell axon terminals. J Neurosci 20(12):4462–4470PubMedGoogle Scholar
  64. Yazulla S, Studholme KM (1992) Light-dependent plasticity of the synaptic terminals of Mb bipolar cells in goldfish retina. J Comp Neurol 320(4):521–530PubMedGoogle Scholar
  65. Yazulla S, Studholme KM (2001) Neurochemical anatomy of the zebrafish retina as determined by immunocytochemistry. J Neurocytol 30(7):551–592PubMedGoogle Scholar
  66. Zhang DR, Yeh HH (1991) Protein kinase C-like immunoreactivity in rod bipolar cells of the rat retina: a developmental study. Vis Neurosci 6(5):429–437PubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Molecular Life SciencesUniversity of ZurichZurichSwitzerland

Personalised recommendations