Skip to main content

Advertisement

SpringerLink
Log in

Studies on retinal mechanisms possibly related to myopia inhibition by atropine in the chicken

  • Basic Science
  • Published:
Graefe's Archive for Clinical and Experimental Ophthalmology Aims and scope Submit manuscript

Abstract

Purpose

While low-dose atropine eye drops are currently widely used to inhibit myopia development in children, the underlying mechanisms are poorly understood. Therefore, we studied possible retinal mechanisms and receptors that are potentially involved in myopia inhibition by atropine.

Methods

A total of 250 μg atropine were intravitreally injected into one eye of 19 chickens, while the fellow eyes received saline and served as controls. After 1 h, 1.5 h, 2 h, 3 h, and 4 h, eyes were prepared for vitreal dopamine (DA) measurements, using high-pressure liquid chromatography with electrochemical detection. Twenty-four animals were kept either in bright light (8500 lx) or standard light (500 lx) after atropine injection for 1.5 h before DA was measured. In 10 chickens, the α2A-adrenoreceptor (α2A-ADR) agonists brimonidine and clonidine were intravitreally injected into one eye, the fellow eye served as control, and vitreal DA content was measured after 1.5 h. In 6 chickens, immunohistochemical analyses were performed 1.5 h after atropine injection.

Results

Vitreal DA levels increased after a single intravitreal atropine injection, with a peak difference between both eyes after 1.97 h. DA was also enhanced in fellow eyes, suggesting a systemic action of intravitreally administered atropine. Bright light and atropine (which both inhibit myopia) had additive effects on DA release. Quantitative immunolabelling showed that atropine heavily stimulated retinal activity markers ZENK and c-Fos in cells of the inner nuclear layer. Since atropine was recently found to also bind to α2A-ADRs at doses where it can inhibit myopia, their retinal localization was studied. In amacrine cells, α2A-ADRs were colocalized with tyrosine hydroxylase (TH), glucagon, and nitric oxide synthase, peptides known to play a role in myopia development in chickens. Intravitreal atropine injection reduced the number of neurons that were double-labelled for TH and α2A-ADR. α2A-ADR agonists clonidine and brimonidine (which were also found by other authors to inhibit myopia) severely reduced vitreal DA content in both injected and fellow eyes, compared to eyes of untreated chicks.

Conclusions

Merging our results with published data, it can be concluded that both muscarinic and α2A-adrenergic receptors are expressed on dopaminergic neurons and both atropine and α2A-ADR antagonists stimulate DA release whereas α2A-ADR agonists strongly suppress its release. Stimulation of DA by atropine was enhanced by bright light. Results are in line with the hypothesis that inhibition of deprivation myopia is correlated with DA stimulation, as long as no toxicity is involved.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Cohn H (1892) Lehrbuch der Hygiene des Auges. Urban & Schwarzenberg Wien Leipzig

  2. McBrien NA, Moghaddam HO, Reeder AP (1993) Atropine reduces experimental myopia and eye enlargement via a nonaccommodative mechanism. Invest Ophthalmol Vis Sci 34:205–215

    CAS  PubMed  Google Scholar 

  3. Carr BJ, Mihara K, Ramachandran R, Saifeddine M, Nathanson NM, Stell WK, Hollenberg D (2018) Myopia-inhibiting concentrations of muscarinic receptor antagonists block activation of alpha2a-adrenoceptors in vitro. Invest Ophthalmol Vis Sci 59:2778–2791

    CAS  PubMed  Google Scholar 

  4. Diether S, Schaeffel F, Lambrou GN, Fritsch C, Trendelenburg AU (2007) Effects of intravitreally and intraperitoneally injected atropine on two types of experimental myopia in chicken. Exp Eye Res 84:266–274

    CAS  PubMed  Google Scholar 

  5. Chia A, Lu QS, Tan D (2016) Five-year clinical trial on atropine for the treatment of myopia 2: myopia control with atropine 0.01% eye drops. Ophthalmol 123:391–399

    Google Scholar 

  6. Pineles SL, Kraker RT, VanderVeen DK, Hutchinson AK, Galvin JA, Wilson LB, Lambert SR (2017) Atropine for the prevention of myopia progression in children: a report by the American Academy of Ophthalmology. Ophthalmol 124:1857–1866

    Google Scholar 

  7. Kradjan WA, Smallridge R, Davis R, Verma P (1985) Atropine serum concentrations after multiple inhaled doses of atropine sulfate. Clin Pharmacol Ther 38:12–15

    CAS  PubMed  Google Scholar 

  8. Fischer AJ, Miethke P, Morgan IG, Stell WK (1998) Cholinergic amacrine cells are not required for the progression and atropine-mediated suppression of form-deprivation myopia. Brain Res 794:48–60

    CAS  PubMed  Google Scholar 

  9. McBrien NA, Arumugam B, Gentle A, Chow A, Sahebjada S (2011) The M4 muscarinic antagonist MT-3 inhibits myopia in chick: evidence for site of action. Ophthalmic Physiol Opt 31:529–523

    PubMed  Google Scholar 

  10. Nickla DL, Yusupova Y, Totonelly K (2015) The muscarinic antagonist MT3 distinguishes between form deprivation- and negative lens-induced myopia in chicks. Curr Eye Res 40:962–967

    CAS  PubMed  Google Scholar 

  11. Carr BJ, Stell WK (2016) Nitric oxide (NO) mediates the inhibition of form-deprivation myopia by atropine in chicks. Sci Rep 6:9

    PubMed  PubMed Central  Google Scholar 

  12. Diether S, Schaeffel F (1999) Long-term changes in retinal contrast sensitivity in chicks from frosted occluders and drugs: relations to myopia? Vis Res 39:2499–2510

    CAS  PubMed  Google Scholar 

  13. Schwahn HN, Kaymak H, Schaeffel F (2000) Effects of atropine on refractive development, dopamine release, and slow retinal potentials in the chick. Vis Neurosci 17:165–176

    CAS  PubMed  Google Scholar 

  14. Nickla DL, Zhu X, Wallman J (2013) Effects of muscarinic agents on chick choroids in intact eyes and eyecups: evidence for a muscarinic mechanism in choroidal thinning. Ophthalmic Physiol Opt 33:245–245

    PubMed  PubMed Central  Google Scholar 

  15. Zhang Z, Zhou Y, Xie Z, Chen T, Gu Y, Lu S, Wu Z (2016) The effect of topical atropine on the choroidal thickness of healthy children. Sci Rep 6:34936

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Chiang ST, Phillips JR (2018) Effect of atropine eye drops on choroidal thinning induced by hyperopic retinal defocus. J Ophthalmol 14:8528315

    Google Scholar 

  17. Wallman J, Wildsoet C, Xu A, Gottlieb MD, Nickla DL, Marran L, Krebs W, Christensen AM (1995) Moving the retina: choroidal modulation of refractive state. Vis Res 35:37–50

    CAS  PubMed  Google Scholar 

  18. Wallman J, Winawer J (2004) Homeostasis of eye growth and the question of myopia. Neuron 43:447–468

    CAS  PubMed  Google Scholar 

  19. Read SA, Alonso-Caneiro D, Vincent SJ, Collins MJ (2015) Longitudinal changes in choroidal thickness and eye growth in childhood. Invest Ophthalmol Vis Sci 56:3103–3112

    PubMed  Google Scholar 

  20. Nickla DL, Totonelly K, Dhillon (2010) Dopaminergic agonists that result in ocular growth inhibition also elicit transient increases in choroidal thickness in chicks. Exp Eye Res 91: 715–720

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Schaeffel F, Howland HC (1991) Properties of the feedback loops controlling eye growth and refractive state in the chicken. Vis Res 31:717–734

    CAS  PubMed  Google Scholar 

  22. Schaeffel F, Bartmann M, Hagel G, Zrenner E (1995) Studies on the role of the retinal dopamine/melatonin system in experimental refractive errors in chickens. Vis Res 35:1247–1264

    CAS  PubMed  Google Scholar 

  23. Li XX, Schaeffel F, Kohler K, Zrenner E (1992) Dose-dependent effects of 6-hydroxy dopamine on deprivation myopia, electroretinograms, and dopaminergic amacrine cells in chickens. Vis Neurosci 9:483–492

    CAS  PubMed  Google Scholar 

  24. Mathis U, Ziemssen F, Schaeffel F (2014) Effects of a human VEGF antibody (bevacizumab) on deprivation myopia and choroidal thickness in the chicken. Exp Eye Res 127:161–169

    CAS  PubMed  Google Scholar 

  25. Megaw PL, Boelen MG, Morgan IG, Boelen MK (2006) Diurnal patterns of dopamine release in chicken retina. Neurochem Int 48:17–23

    CAS  PubMed  Google Scholar 

  26. Carr BJ, Nguyen CT, Stell WK (2019) Alpha2 -adrenoceptor agonists inhibit form-deprivation myopia in the chick. Clin Exp Optom 102:418–425

    PubMed  PubMed Central  Google Scholar 

  27. Mathis U, Schaeffel F (2010) Transforming growth factor-beta in the chicken fundal layers: an immunohistochemical study. Exp Eye Res 90:780–790

    CAS  PubMed  Google Scholar 

  28. Fischer AJ, Reh TA (2000) Identification of a proliferating marginal zone of retinal progenitors in postnatal chickens. Dev Biol 220:197–210

    CAS  PubMed  Google Scholar 

  29. Bitzer M, Schaeffel F (2002) Defocus-induced changes in ZENK expression in the chicken retina. Invest Ophthalmol Vis Sci 43:246–252

    PubMed  Google Scholar 

  30. Edqvist PH, Myers SM, Hallböök F (2006) Early identification of retinal subtypes in the developing, pre-laminated chick retina using the transcription factors Prox1, Lim1, Ap2alpha, Pax6, Isl1, Isl2, Lim3 and Chx10. Eur J Histochem 50:147–154

    CAS  PubMed  Google Scholar 

  31. Fischer AJ, Stanke JJ, Aloisio G, Hoy H, Stell WK (2007) Heterogeneity of horizontal cells in the chicken retina. J Comp Neurol 500:1154–1171

    CAS  PubMed  Google Scholar 

  32. Fischer AJ, Schmidt M, Omar G, Reh TA (2004) BMP4 and CNTF are neuroprotective and suppress damage-induced proliferation of Müller glia in the retina. Mol Cell Neurosci 27:531–542

    CAS  PubMed  Google Scholar 

  33. Bitzer M, Kovacs B, Feldkaemper M, Schaeffel F (2006) Effects of muscarinic antagonists on ZENK expression in the chicken retina. Exp Eye Res 8:379–388

    Google Scholar 

  34. Harun-Or-Rashid M, Lindqvist N, Hallböök F (2014) Transactivation of EGF receptors in chicken Müller cells by α2A-adrenergic receptors stimulated by brimonidine. Invest Ophthalmol Vis Sci 55:3385–3394

    CAS  PubMed  Google Scholar 

  35. Fischer AJ, Stell WK (1999) Nitric oxide synthase-containing cells in the retina, pigmented epithelium, choroid, and sclera of the chick eye. J Comp Neurol 405:1–14

    CAS  PubMed  Google Scholar 

  36. Bitzer M, Schaeffel F (2004) Effects of quisqualic acid on retinal ZENK expression induced by imposed defocus in the chick eye. Optom Vis Sci 81:127–136

    PubMed  Google Scholar 

  37. Ohngemach S, Hagel G, Schaeffel F (1997) Concentrations of biogenic amines in fundal layers in chickens with normal visual experience, deprivation, and after reserpine application. Vis Neurosci 14:493–505

    CAS  PubMed  Google Scholar 

  38. Rohrer B, Iuvone PM, Stell WK (1995) Stimulation of dopaminergic amacrine cells by stroboscopic illumination or fibroblast growth factor (bFGF, FGF-2) injections: possible roles in prevention of form deprivation myopia in the chick. Brain Res 686:169–181

    CAS  PubMed  Google Scholar 

  39. Luft WA, Iuvone PM, Stell WK (2004) Spatial, temporal, and intensive determinants of dopamine release in the chick retina. Vis Neurosci 21:627–635

    CAS  PubMed  Google Scholar 

  40. Simon P, Feldkaemper M, Bitzer M, Ohngemach S, Schaeffel F (2004) Early transcriptional changes of retinal and choroidal TGFbeta-2, RALDH-2, and ZENK following imposed positive and negative defocus in chickens. Mol Vis 10:588–597

    CAS  PubMed  Google Scholar 

  41. Araki CM, Hamassaki-Britto DE (1998) Motion-sensitive neurons in the chick retina: a study using Fos immunohistochemistry. Brain Res 794:333–337

    CAS  PubMed  Google Scholar 

  42. Su YY, Watt CB (1987) Interaction between enkephalin and dopamine in the avian retina. Brain Res 423:63–70

    CAS  PubMed  Google Scholar 

  43. Yamagata K, Goto K, Kuo CH, Kondo H, Miki N (1990) Visinin: a novel calcium binding protein expressed in retinal cone cells. Neuron 4:469–476

    CAS  PubMed  Google Scholar 

  44. Bruhn SL, Cepko CL (1996) Development of the pattern of photoreceptors in the chick retina. Neurosci 16:1430–1439

    CAS  Google Scholar 

  45. Fischer AJ, Foster S, Scott MA, Sherwood P (2008) Transient expression of LIM-domain transcription factors is coincident with delayed maturation of photoreceptors in the chicken retina. J Comp Neurol 506:584–603

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Ellis JH, Richards DE, Rogers JH (1991) Calretinin and calbindin in the retina of the developing chicken. Cell Tissue Res 264:197–208

    CAS  PubMed  Google Scholar 

  47. Iuvone PM, Galli CL, Garrison-Gund CK, Neff NH (1978) Light stimulates tyrosine hydroxylase activity and dopamine synthesis in retinal amacrine neurons. Science 202:901–902

    CAS  PubMed  Google Scholar 

  48. Stone RA, Lin T, Laties AM, Iuvone PM (1989) Retinal dopamine and form-deprivation myopia. Proc Natl Acad Sci 86:704–706

    CAS  PubMed  Google Scholar 

  49. Fischer AJ, McGuire JJ, Schaeffel F, Stell WK (1999) Light- and focus-dependent expression of the transcription factor ZENK in the chick retina. Nat Neurosci 2:706–712

    CAS  PubMed  Google Scholar 

  50. Vessey KA, Lencses KA, Rushforth DA, Hruby VJ, Stell WK (2005) Glucagon receptor agonists and antagonists affect the growth of the chick eye: a role for glucagonergic regulation of emmetropization? Invest Ophthalmol Vis Sci 46:3922–3931

    PubMed  PubMed Central  Google Scholar 

  51. Fischer AJ, McKinnon L, Nathanson NM, Stell WK (1998) Identification and localization of muscarinic acetylcholine receptors in the ocular tissues of the chick. J Comp Neurol 392:273–284

    CAS  PubMed  Google Scholar 

  52. Stanke JJ, Lehman B, Fischer AJ (2008) Muscarinic signaling influences the patterning and phenotype of cholinergic amacrine cells in the developing chick retina. BMC Dev Biol 8:13

    PubMed  PubMed Central  Google Scholar 

  53. Fischer AJ, Seltner RL, Poon J, Stell WK (1998) Immunocytochemical characterization of quisqualic acid- and N-methyl-D-aspartate-induced excitotoxicity in the retina of chicks. J Comp Neurol 393:1–15

    CAS  PubMed  Google Scholar 

  54. Iuvone PM, Rauch AL (1983) Alpha 2-adrenergic receptors influence tyrosine hydroxylase activity in retinal dopamine neurons. Life Sci 33:2455–2463

    CAS  PubMed  Google Scholar 

  55. Zhou X, Pardue MT, Iuvone PM, Qu J (2017) Dopamine signaling and myopia development: what are the key challenges. Prog Retin Eye Res 61:60–71

    CAS  PubMed  PubMed Central  Google Scholar 

  56. Liu Y, Wang Y, Lv H, Jiang X, Zhang M, Li X (2017) α-Adrenergic agonist brimonidine control of experimentally induced myopia in Guinea pigs: a pilot study. Mol Vis 23:785 798. eCollection

  57. Myhr KL, McReynolds JS (1996) Cholinergic modulation of dopamine release and horizontal cell coupling in mudpuppy retina. Vis Res 36:3933–3938

    CAS  PubMed  Google Scholar 

  58. Hare WA, Owen WG (1995) Similar effects of carbachol and dopamine on neurons in the distal retina of the tiger salamander. Vis Neurosci 12:443–455

    CAS  PubMed  Google Scholar 

  59. Luft WA, Ming Y, Stell WK (2003) Variable effects of previously untested muscarinic receptor antagonists on experimental myopia. Invest Ophthalmol Vis Sci 44:1330–1338

    PubMed  Google Scholar 

  60. Dubocovich ML (1984) Alpha-2 adrenoceptors modulate [3H]dopamine release from rabbit retina. J Pharmacol Exp Ther 230:149–155

    CAS  PubMed  Google Scholar 

  61. Ashby R, Ohlendorf A, Schaeffel F (2009) The effect of ambient illuminance on the development of deprivation myopia in chicks. Invest Ophthalmol Vis Sci 50:5348–5354

    PubMed  Google Scholar 

  62. Ashby RS, Schaeffel F (2010) The effect of bright light on lens compensation in chicks. Invest Ophthalmol Vis Sci 51:5247–5253

    PubMed  Google Scholar 

  63. Karouta C, Ashby RS (2014) Correlation between light levels and the development of deprivation myopia. Invest Ophthalmol Vis Sci 56:299–309

    PubMed  Google Scholar 

  64. Lan W, Feldkaemper M, Schaeffel F (2013) Bright light induces choroidal thickening in chickens. Optom Vis Sci 90:1199–1206

    PubMed  Google Scholar 

  65. Lan W, Feldkaemper M, Schaeffel F (2014) Intermittent episodes of bright light suppress myopia in the chicken more than continuous bright light. PLoS One 9:e110906

    PubMed  PubMed Central  Google Scholar 

  66. Lan W, Yang Z, Feldkaemper M, Schaeffel F (2016) Changes in dopamine and ZENK during suppression of myopia in chicks by intense illuminance. Exp Eye Res 145:118–124

    CAS  PubMed  Google Scholar 

  67. Norton TT (2016) What do animal studies tell us about the mechanism of myopia-protection by light? Optom Vis Sci 93:1049–1051

    PubMed  PubMed Central  Google Scholar 

  68. Smith EL 3rd, Hung LF, Huang J (2012) Protective effects of high ambient lighting on the development of form-deprivation myopia in rhesus monkeys. Invest Ophthalmol Vis Sci 53:421–428

    PubMed  PubMed Central  Google Scholar 

  69. Smith EL 3rd, Hung LF, Arumugam B, Huang J (2013) Negative lens-induced myopia in infant monkeys: effects of high ambient lighting. Invest Ophthalmol Vis Sci 26:2959–2969

    Google Scholar 

  70. Rose KA, Morgan IG, Ip J, Kifley A, Huynh S, Smith W, Mitchell P (2008) Outdoor activity reduces the prevalence of myopia in children. Ophthalmology 115:1279–1285

    PubMed  Google Scholar 

  71. Boelen MK, Boelen MG, Marshak DW (1998) Light-stimulated release of dopamine from the primate retina is blocked by 1-2-amino-4-phosphonobutyric acid (APB). Vis Neurosci 15(97):103

    Google Scholar 

  72. McCarthy CS, Megaw P, Devadas M, Morgan IG (2007) Dopaminergic agents affect the ability of brief periods of normal vision to prevent form-deprivation myopia. Exp Eye Res 84:100–107

    CAS  PubMed  Google Scholar 

  73. Megaw P, Morgan I, Boelen MJ (2001) Vitreal dihydroxyphenylacetic acid (DOPAC) as an index of retinal dopamine release. Neurochem 76:1636–1644

    CAS  Google Scholar 

  74. Iuvone PM, Tigges M, Stone RA, Lambert S, Laties AM (1991) Effects of apomorphine, a dopamine receptor agonist, on ocular refraction and axial elongation in a primate model of myopia. Invest Ophthalmol Vis Sci 32:1674–1677

    CAS  PubMed  Google Scholar 

  75. Chen S, Zhi Z, Ruan Q, Liu Q, Li F, Wan F, Reinach PS, Chen J, Qu J, Zhou X (2017) Bright light suppresses form-deprivation myopia development with activation of dopamine D1 receptor signaling in the ON pathway in retina. Invest Ophthalmol Vis Sci 58:2306–2316

    CAS  PubMed  Google Scholar 

  76. Wulle I, Kirsch M, Wagner HJ (1990) Cyclic changes in dopamine and DOPAC content, and tyrosine hydroxylase activity in the retina of a cichlid fish. Brain Res 515:163–167

    CAS  PubMed  Google Scholar 

  77. Kolbinger W, Weiler R (1993) Modulation of endogenous dopamine release in the turtle retina: effects of light, calcium, and neurotransmitters. Vis Neurosci 10:1035–1041

    CAS  PubMed  Google Scholar 

  78. Boatright JH, Gordon JR, Iuvone PM (1994) Inhibition of endogenous dopamine release in amphibian retina by L-2-amino-4-phosphonobutyric acid (L-AP4) and trans-2 aminocyclopentane-1,3-dicarboxylate (ACPD). Brain Res 649:339–342

    CAS  PubMed  Google Scholar 

  79. Morgan IG, Boelen MK (1996) A retinal dark-light switch: a review of the evidence. Vis Neurosci 13:399–409

    CAS  PubMed  Google Scholar 

  80. Feldkaemper M, Schaeffel F (2013) An updated view on the role of dopamine in myopia. Exp Eye Res 114:106–119

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Sandra Bernhard-Kurz for excellent technical assistance.

Funding

This study was supported by the German Research Council (DFG Scha 518/15–1) to UM and FS, the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie Research Training Network MyFun Grant MSCA-ITN-2015-675137 to FS and MF, and a stipend from the China Scholarship Council (No. 201606370188) to MW.

Author information

Authors and Affiliations

  1. Section of Neurobiology of the Eye, Ophthalmic Research Institute, University of Tuebingen, Tuebingen, Germany

    Ute Mathis, Marita Feldkaemper, Min Wang & Frank Schaeffel

Authors
  1. Ute Mathis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marita Feldkaemper
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Min Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Frank Schaeffel
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Frank Schaeffel.

Ethics declarations

Ute Mathis declares that she has no conflict of interest. Marita Feldkaemper declares that she has no conflict of interest. Min Wang declares that she has no conflict of interest. Frank Schaeffel declares that he has no conflict of interest. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mathis, U., Feldkaemper, M., Wang, M. et al. Studies on retinal mechanisms possibly related to myopia inhibition by atropine in the chicken. Graefes Arch Clin Exp Ophthalmol 258, 319–333 (2020). https://doi.org/10.1007/s00417-019-04573-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00417-019-04573-y

Keywords

Search

Navigation