Advertisement

Cell and Tissue Research

, Volume 315, Issue 2, pp 197–201 | Cite as

Localization of Aa-nat mRNA in the rat retina by fluorescence in situ hybridization and laser capture microdissection

  • Cuimei Liu
  • Chiaki Fukuhara
  • James H. WesselIII
  • P. Michael Iuvone
  • Gianluca Tosini
Regular Article

Abstract

Arylalkylamine N-acetyltransferase (AA-NAT) is the key regulatory enzyme in the melatonin biosynthetic pathway. Previous investigations have reported that Aa-nat mRNA in rat is only detected in a sub-population of photoreceptor cells that resemble cones in shape and size. In the present study, we investigated Aa-nat expression in the rat retina by using in situ hybridization and laser capture microdissection combined with the reverse transcription/polymerase chain reaction technique. Our results demonstrate that, contrary to previous reports, Aa-nat transcripts are present not only in the photoreceptor cells, but also in the inner nuclear layer and in the ganglion cell layer. However, the rhythmic expression of Aa-nat mRNA was observed only in photoreceptor cells.

Keywords

Retina Melatonin Circadian rhythm Aa-nat Photoreceptors Ganglion cells Rat (Wistar, adult male) 

References

  1. Bernard M, Iuvone PM, Cassone VM, Roseboom PH, Coon PL, Klein DC (1997) Avian melatonin synthesis: photic and circadian regulation of serotonin N-acetyltransferase mRNA in the chicken pineal gland and retina. J Neurochem 68:213–224PubMedGoogle Scholar
  2. Blazynski C, Dubocovich ML (1991) Localization of 2-[125I]iodomelatonin binding sites in mammalian retina. J Neurochem 56:1873–1880PubMedGoogle Scholar
  3. Cassone VM (1990) Effects of melatonin on vertebrate circadian system. Trends Neurosci 13:457–464PubMedGoogle Scholar
  4. Coon SL, Del Olmo E, Young WS, Klein DC (2002) Melatonin synthesis enzymes in Macaca mulatta: focus on arylalkylamine N-acetyltransferase (EC 2.3.1.87). J Clin Endocrinol Metab 87:4699–4706CrossRefPubMedGoogle Scholar
  5. Eberwine JH, Valentino KL, Barchas JD (1994) In situ hydridization in neurobiology: advances in methodology, Oxford University Press, New YorkGoogle Scholar
  6. Fujieda H, Hamadanizadeh SA, Wankiewicz E, Pang SF, Brown GM (1999) Expression of mt1 melatonin receptor in rat retina: evidence for multiple cell targets for melatonin. Neuroscience 93:793–799CrossRefPubMedGoogle Scholar
  7. Fukuhara C, Dirden JC, Tosini G (2001) Photic regulation of melatonin in rat retina and the role of proteasomal proteolysis. Neuroreport 12:3833–3837PubMedGoogle Scholar
  8. Guido M, Garbarino Pico E, Keller-Sarmiento MI, Rosenstein RE, Caputto BL (2000) Circadian rhythmicity in retinal ganglion cells. Soc Neurosci Abs 26:208Google Scholar
  9. Niki T, Hamada T, Ohtomi M, Sakamoto K, Suzuki S, Kako K, Hosoya Y, Horikawa K, Ishida N (1998) The localization of the site of arylalkylamine N-acetyltransferase circadian expression in the photoreceptor cells of mammalian retina. Biochem Biophys Res Commun 248:115–120PubMedGoogle Scholar
  10. Sakamoto K, Ishida N (1998a) Circadian expression of serotonin N-acetyltransferase mRNA in the rat retina. Neurosci Lett 245:113–116PubMedGoogle Scholar
  11. Sakamoto K, Ishida K (1998b) Molecular cloning of serotonin N-acetyltransferase gene from the mouse and its daily expression in the retina. Neurosci Lett 250:181–184PubMedGoogle Scholar
  12. Sakamoto K, Oishi K, Shiraishi M, Hamano S, Otsuka H, Miyake Y, Ishida N (2000) Two circadian oscillatory mechanisms in the mammalian retina. Neuroreport 11:3995–3997PubMedGoogle Scholar
  13. Tosini G (2000) Melatonin circadian rhythm in the retina of mammals. Chronobiol Int 17: 599–612PubMedGoogle Scholar
  14. Tosini G, Fukuhara C (2002) The mammalian retina as a clock. Cell Tissue Res 309:119–126CrossRefPubMedGoogle Scholar
  15. Tosini G, Menaker M (1996) Circadian rhythms in cultured mammalian retina. Science 272:419–421PubMedGoogle Scholar
  16. Tosini G, Menaker M (1998) The clock in the mouse retina: melatonin synthesis and photoreceptor degeneration. Brain Res 789:221–228PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Cuimei Liu
    • 1
  • Chiaki Fukuhara
    • 1
  • James H. WesselIII
    • 2
  • P. Michael Iuvone
    • 2
  • Gianluca Tosini
    • 1
  1. 1.Neuroscience Institute and NSF Center for Behavioral NeuroscienceMorehouse School of MedicineAtlantaUSA
  2. 2.Department of PharmacologyEmory University School of MedicineAtlantaUSA

Personalised recommendations