Abstract
The distribution of neuronal nitric oxide synthase (NOS) immunoreactivity was examined in rat and rabbit retinas and was compared with the distribution of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase reactivity and vasoactive intestinal peptide (VIP) immunoreactivity. An antibody raised against a C-terminal fragment of a cloned rat cerebellar NOS was used to localise NOS immunoreactivity. NOS immunoreactive cells were not detected in rat retinas at postnatal day 1 or 4, but were seen from postnatal day 7 onwards. NOS immunolabelling was seen in a small population of cells in the proximal inner nuclear layer. Most of the labelled cells had the position of amacrine cells and were seen to send processes into the inner plexiform layer. A few labelled cells were at times also seen in the ganglion cell layer, which are likely to correspond to displaced amacrine cells. The same NOS-labelling pattern was seen in rat and rabbit retinas.
NADPH-diaphorase staining was observed in both species, in photoreceptor inner segments, in cells with the position of horizontal cells, in a subset of amacrine and displaced amacrine cells, in large cell bodies in the ganglion cell layer, in both plexiform layers, and in endothelium. Colocalisation of NOS immunoreactivity and NADPH-diaphorase staining was only observed among amacrine cells. However, not all NADPH-diaphorase-reactive amacrine cells were found to be NOS immunoreactive. VIP immunoreactivity was also localised in rat retinas in a subpopulation of amacrine cells, but no colocalisation of NOS and VIP immunoreactivity was observed.
Our observations indicate that only amacrine cells contain the NOS form recognisable by the antibody used, and suggest that different isoforms of neuronal NOS may be present in retinal cells. Further, the onset of NOS expression in rat amacrine cells appears to occur independently of neuronal activity.
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References
Ahmad I, Barnstable CJ (1993) Differential laminar expression of particulate and soluble guanylate cyclase genes in rat retina. Exp Eye Res 56:51–62
Ahmad I, Leinders-Zufall T, Kocsis J, Shepherd GM, Zufall F, Barnstable CJ (1994) Retinal ganglion cells express a cGMP-gated cation conductance activatable by nitric oxide donors. Neuron 12:155–165
Alm P, Larsson B, Ekblad E, Sundler F, Andersson K-E (1993) Immunohistochemical localization of peripheral nitric oxide synthase-containing nerves using antibodies raised against synthesized C- and N-terminal fragments of a cloned enzyme from rat brain. Acta Physiol Scand 148:421–429
Bredt DS, Glatt CE, Hwang PM, Fotuhi M, Dawson TM, Snyder SH (1991) Nitric oxide synthase protein and mRNA are discretely localised in neuronal populations of the mammalian CNS together with NADPH diaphorase. Neuron 7:615–624
Bugnon O, Schaad NC, Schorderet M (1994) Nitric oxide modulates endogenous dopamine release in bovine retina. Neuroreport 5:401–404
Dawson TM, Bredt DS, Fotuhi M, Hwang PM, Snyder SH (1991) Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues. Proc Natl Acad Sci USA 88:7797–7801
Förstermann U, Schmidt HHHW, Pollock JS, Sheng H, Mitchell JA, Warner TD, Nakane M, Murad F (1991) Isoforms of nitric oxide synthase. Characterization and purification from different cell types. Biochem Pharmacol 42:1849–1857
Grozdanovic Z, Baumgarten HG, Brüning G (1992) Histochemistry of NADPH-diaphorase, a marker for neuronal nitric oxide synthase, in the peripheral autonomic nervous system of the mouse. Neuroscience 48:225–235
Grün G (1982) The development of the vertebrate retina: a comparative survey. In: Beck F, Hild W, van Limborgh J, Ortmann R, Pauly JE, Schiebler TH (eds) Advances in anatomy embryology and cell biology, vol 78. Springer, Berlin Heidelberg New York, pp 1–85
Hope BT, Michael GJ, Knigge KM, Vincent SR (1991) Neuronal NADPH diaphorase is a nitric oxide synthase. Proc Natl Acad Sci USA 88:2811–2814
Horio Y, Murad F (1991) Solubilization of guanylyl cyclase from bovine rod outer segments and effects of lowering Ca2+ and nitro compounds. J Biol Chem 266:3411–3415
Knowles RG, Palacios M, Palmer RMJ, Moncada S (1989) Formation of nitric oxide from L-arginine in the central nervous system: a transduction mechanism for stimulation of the soluble guanylate cyclase. Proc Natl Acad Sci USA 86:5159–5162
Koch K-W, Lambrecht H-G, Haberecht M, Redburn D, Schmidt HHHW (1994) Functional coupling of a Ca2+/calmodulin-dependent nitric oxide synthase and a soluble guanylyl cyclase in vertebrate photoreceptor cells. EMBO J 13:3312–3320
Mitrofanis J (1989) Development of NADPH-diaphorase cells in the rat's retina. Neurosci Lett 102:165–172
Mitrofanis J, Robinson SR, Ashwell K (1992) Development of catecholaminergic, indoleamine-accumulating and NADPH-diaphorase amacrine cells in rabbit retinae. J Comp Neurol 319:560–585
Miyachi E-I, Miyakawa A, Murakami M (1991) Modulation of electrical coupling between retinal horizontal cells by intracellular messengers. Neurosci Res [Suppl 15]:S41–S49
Moncada S (1992) Thel-arginine:nitric oxide pathway. Acta Physiol Scand 145:201–227
Moncada S, Palmer RMJ, Higgs EA (1991) Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 43:109–142
Osborne NN, Barnett NL, Herrera AJ (1993) NADPH diaphorase localization and nitric oxide synthetase activity in the retina and anterior uvea of the rabbi retina. Brain Res 610:194–198
Pozdnyakov N, Lloyd A, Reddy VN, Sitaramayya A (1993) Nitric oxide-regulated endogenous ADP-ribosylation of rod outer segment proteins. Biochem Biophys Res Commun 192:610–615
Sagar SM (1986) NADPH diaphorase histochemistry in the rabbit retina. Brain Res 373:153–158
Sagar SM (1990) NADPH-diaphorase reactive neurons of the rabbit retina: differential sensitivity to excitotoxins and unusual morphological features. J Comp Neurol 300:309–319
Sandell JH (1985) NADPH diaphorase cells in the mammalian inner retina. J Comp Neurol 238:466–472
Sato T (1990) NADPH-diaphorase positive amacrine cells in the retinae of the frog (Rana esculenta) and pigeon (Columbia livia). Arch Histol Cytol 53:63–69
Schmidt K-F, Nöll GN, Yamamoto Y (1992) Sodium nitroprusside alters dark voltage and light responses in isolated retinal rods during whole-cell recording. Vis Neurosci 9:205–209
Schuman EM, Madison DV (1994) Nitric oxide and synaptic function. Annu Rev Neurosci 17:153–183
Shiells R and Falk G (1992) Retinal on-bipolar cells contain a nitric oxide-sensitive guanylate cyclase. Neuroreport 3:845–848
Straznicky C, Gábriel R (1991) NADPH-diaphorase positive neurons in the retina of Bufo marinus: selective staining of bipolar and amacrine cells. Arch Histol Cytol 54:213–220
Ujihara H, Akaike A, Tamura Y, Yokota T, Sasa M, Kashii S, Honda Y (1993) Blockade of retinal NMDA receptors by sodium nitroprusside is probably due to nitric oxide formation. Jpn J Pharmacol 61:375–377
Vaccaro TM, Cobcroft MD, Provis JM, Mitrofanis J (1991) NADPH-diaphorase reactivity in adult and developing cat retinae. Cell Tissue Res 265:371–379
Vaney DI, Young HM (1988) GABA-like immunoreactivity in ADPH-diaphorase amacrine cells of the rabbit retina. Brain Res 474:380–385
Venturini CM, Knowles RG, Palmer RMJ, Moncada S (1991) Synthesis of nitric oxide in the bovine retina. Biochem Biophys Res Commun 180:920–925
Vincent SR, Kimura H (1992) Histochemical mapping of nitric oxide synthase in the rat brain. Neuroscience 46:755–784
Wässle H, Chun MH, Müller F (1987) Amacrine cells in the ganglion cell layer of the cat retina. J Comp Neurol 265:391–408
Weidman TA, Kuwabara T (1968) Postnatal development of the rat retina. An electron microscopic study. Arch Ophthalmol 79:470–484
Yamamoto R, Bredt DS, Snyder SH, Stone RA (1993a) The localization of nitric oxide synthase in the rat eye and related cranial ganglia. Neuroscience 54:189–200
Yamamoto R, Bredt DS, Dawson TM, Snyder SH, Stone RA (1993b) Enhanced expression of nitric oxide synthase by rat retina following pterygopalatine parasympathetic denervation. Brain Res 631:83–88
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Paper in honour of Professor Rolf Elofsson on the occasion of his retirement from the chair of Zoology at the University of Lund
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Perez, M.T.R., Larsson, B., Alm, P. et al. Localisation of neuronal nitric oxide synthase-immunoreactivity in rat and rabbit retinas. Exp Brain Res 104, 207–217 (1995). https://doi.org/10.1007/BF00242007
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DOI: https://doi.org/10.1007/BF00242007