The effect of experimental ischaemia and excitatory amino acid agonists on the GABA and serotonin immunoreactivities in the rabbit retina.
The aim of the described experiments was to use immunohistochemistry to visualize the release of GABA from specific retinal amacrine cells following ischaemia and to establish the involvement of defined glutamatergic receptors. In initial experiments, rabbit retinas were exposed in vitro to excitatory amino acid agonists alone or in combination with a putative antagonist, or in physiological solution lacking oxygen and glucose, or in solution containing potassium cyanide for 45 min at 37 degrees C. The nature of the GABA immunoreactivity was then examined by immunohistochemistry. In other in vitro experiments, retinas were first allowed to accumulate exogenous serotonin before exposing the tissues to the combinations as described. These tissues were then processed immunohistochemically for the localization of serotonin. In yet other experiments, the intraocular pressure of a rabbit's eye was raised to about 110 mmHg for 60 min and a reperfusion time of 45 min allowed before dissecting the retina and processing for the localization of GABA immunoreactivity. The other eye served as a control. Of the excitatory amino acid agonists tested, only N-methyl-D-aspartate, kainate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid caused a change in the GABA immunoreactivity. The N-methyl-D-aspartate effect was specifically antagonized by dizocilpine maleate, dextromethorphan and memantine, and was characterized by a reduction in the number of GABA-immunoreactive perikarya. The GABA "staining" in the inner plexiform layer also appeared as four clear bands. The alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid- and kainate-induced effects were both antagonized by 6-cyano-2,3-dihydroxy-7-nitroquinoxaline-2,3-dione and partially by kynurenic acid at the concentrations used. Here, the amount of GABA-positive perikarya was greatly reduced and three immunoreactive bands appeared in the inner plexiform layer. However, for low concentrations of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid four GABA-immunoreactive bands could be identified in the inner plexiform layer. The normal GABA immunoreactivity of the inner plexiform layer also appeared to be in defined bands in retinas which received an ischaemic insult either by reducing the availability of glucose and oxygen, exposing the tissue to potassium cyanide or raising the intraocular pressure of an eye. In these cases the number of GABA-positive perikarya was also reduced. Only alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and kainate of the excitatory amino acid agonists tested caused a release of serotonin and this process was antagonized by 6-cyano-2,3-dihydroxy-7-nitroquinoxaline-2,3-dione and partially by kynurenic acid.(ABSTRACT TRUNCATED AT 400 WORDS)
Osborne
N N
NN
Nuffield Laboratory of Ophthalmology, University of Oxford, U.K.
Herrera
A J
AJ
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D000613:Aminobutyrates / Q000494:pharmacology; D000818:Animals; D003515:Cycloleucine / Q000031:analogs & derivatives / Q000494:pharmacology; D003915:Dextromethorphan / Q000494:pharmacology; D005136:Eye Proteins / Q000187:drug effects / Q000502:physiology*; D005947:Glucose / Q000494:pharmacology; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D007429:Intraocular Pressure; D007511:Ischemia / Q000378:metabolism*; D007608:Kainic Acid / Q000494:pharmacology*; D007736:Kynurenic Acid / Q000494:pharmacology; D008559:Memantine / Q000494:pharmacology; D016202:N-Methylaspartate / Q000494:pharmacology*; D010100:Oxygen / Q000494:pharmacology; D011190:Potassium Cyanide / Q000633:toxicity; D011810:Quinoxalines / Q000494:pharmacology; D011817:Rabbits; D018091:Receptors, AMPA / Q000187:drug effects / Q000502:physiology; D017470:Receptors, Glutamate / Q000145:classification / Q000187:drug effects / Q000502:physiology*; D018092:Kainic Acid Receptors / Q000187:drug effects / Q000502:physiology; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000502:physiology; D012160:Retina / Q000378:metabolism*; D012171:Retinal Vessels; D012701:Serotonin / Q000378:metabolism* / Q000494:pharmacology; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D000613:Aminobutyrates; D003515:Cycloleucine; D003915:Dextromethorphan; D005136:Eye Proteins; D005947:Glucose; D005971:Glutamates; D018698:Glutamic Acid; D007608:Kainic Acid; D007736:Kynurenic Acid; D008559:Memantine; D016202:N-Methylaspartate; D010100:Oxygen; D011190:Potassium Cyanide; D011810:Quinoxalines; D018091:Receptors, AMPA; D017470:Receptors, Glutamate; D018092:Kainic Acid Receptors; D016194:Receptors, N-Methyl-D-Aspartate; D012701:Serotonin; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; D005680:gamma-Aminobutyric Acid; C036361:1-amino-1,3-dicarboxycyclopentane; C012729:2-amino-4-phosphonobutyric acid