Differential expression of GAD(65) and GAD(67) during the development of the rat retina.

The development of synthetic enzymes in the GABAergic system (GAD(67) and GAD(65)) of the rat retina was analyzed from birth to the 4th postnatal week by the reverse transcriptase polymerase chain reaction (RT-PCR) and by immunohistochemistry. As previously observed for GABA, immunoreactive GAD(67) profiles are seen clearly in the inner retinal layers at birth. At the end of the 1st week of postnatal life, immunolabeling is detected in amacrine and/or ganglion cells and in horizontal cells. GAD(67) immunoreactivity is transiently expressed in horizontal cells and disappears during the 3rd postnatal week. GAD(65) however does not develop until the 5th postnatal day. Immunolabeling is detected in the processes layering the inner plexiform layer (IPL) before being detected in the amacrine and/or ganglion cell bodies. The appearance of transcripts for GAD coincided with the appearance of the proteins. A transient form of mRNA transcripts of the GAD(67) gene containing an extra exon (ES-exon) is also observed which disappears progressively from birth to the 4th postnatal week. This form synthesizes a truncated, enzymatically inactive protein, which could participate in the regulation of GABA synthesis from glutamate present at high levels during retinogenesis.

Retinal TUNEL-positive cells and high glutamate levels in vitreous humor of mutant quail with a glaucoma-like disorder.

PURPOSE: To investigate whether retinal cell death observed in an avian glaucoma-like disorder occurs by apoptosis and whether an increase in excitotoxic amino acid concentration in the vitreous humor is associated temporally with cell death in the retina. METHODS: Presumptive retinal apoptotic nuclei were identified by histochemical detection of DNA fragmentation (by TdT-dUTP terminal nick-end labeling [TUNEL]), and vitreal concentrations of glutamate and several other amino acids were determined by high-pressure liquid chromatography with fluorometric detection in the al mutant quail (Coturnix coturnix japonica) in which a glaucoma-like disorder develops spontaneously. RESULTS: TUNEL-labeled nuclei were located mostly in the ganglion cell layer (GCL) in the retina of mutant quails 3 months after hatching. However, labeled nuclei were also observed in the inner and outer nuclear layers. At 7 months, most TUNEL-positive nuclei were detected in the inner nuclear layer, whereas labeled cells in the GCL were reduced in number. No TUNEL-labeled nuclei were detected in the retina of control quails at any age. Vitreal concentrations of glutamate and aspartate were significantly increased in 1-month-old mutant quails compared with control animals. Concentrations decreased at 3 months, and no significant differences were observed between strains at 7 months. CONCLUSIONS: Presumptive apoptotic cell death is detected from 3 months after hatching in mutant quails and is not restricted to retinal ganglion cells. Cell death appears just after a significant increase in excitotoxic amino acid concentrations in the vitreous humor, suggesting a correlation between both events.

Day and night dysfunction in intraretinal melatonin and related indoleamines metabolism, correlated with the development of glaucoma-like disorder in an avian model.

As previous studies have suggested that melatonin and serotonin may be involved in the regulation of intraocular pressure, retinal concentrations of melatonin, 5-HT, and related indoleamines measured at day and at night were studied during the development of a glaucoma-like disorder with increased intraocular pressure in the al mutant quail. Indoleamine levels were determined by HPLC with electrochemical detection in 1-month-, 3-month-, and 7-month-old al mutant and control quails. Morphology and numbers of melatonin-synthesizing and 5-HT-containing cells, labelled immunohistochemically with an anti-hydroxyindol-0-methyltransferase (HIOMT) antibody and an anti-5-HT antibody, respectively, were studied. Major findings were that: (1) no significant changes in morphology of melatonin-synthesizing cells or in the morphology and density of 5-HT-containing amacrine cells were observed during the development of glaucoma: (2) 5-HT metabolism was modified during the night at 1 month of age and during the day after 3 months; and (3) melatonin metabolism was modified during the night at 7 months and during the day after 3 months. These results demonstrate a relationship between the temporal evolution of this avian glaucoma and a dysfunction in indoleamine retinal metabolism.

Changes in retinal dopaminergic cells and dopamine rhythmic metabolism during the development of a glaucoma-like disorder in quails.

PURPOSE: To examine the possible correlation between a dysfunction of the daily rhythm of retinal dopamine (DA) and the development of a glaucoma-like disorder in an animal model, the al mutant quail (Coturnix coturnix japonica). METHODS: The morphology and density of DA-containing cells labeled immunohistochemically with an anti-tyrosine hydroxylase (TH) antibody were correlated with the diurnal and nocturnal contents of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) determined by high-pressure liquid chromatography with electrochemical detection. RESULTS: The number of TH-immunoreactive cells was lower than normal in mutant quails suffering from the disorder. There were considerably fewer cells in the central retina, and the DA metabolism was reduced in parallel. The nocturnal DA content was lower than the diurnal level in normal quails, but there was no such circadian fluctuation in mutant quails. CONCLUSIONS: This glaucoma-like disorder in quails is correlated with the degeneration of DA-containing amacrine cells and a dysfunction of the circadian rhythmicity of DA synthesis.

Retinal distribution of tyrosine hydroxylase immunoreactive cells in two strains of quails Coturnix coturnix japonica.

We have compared the distribution of tyrosine-hydroxylase immunoreactive (TH-I) somata in the retina of two strains of japanese quails, one normally pigmented and one hypopigmented mutant which develops a glaucoma, three months after their hatching. TH-I cells were observed among amacrine and presumptive interplexiform cells in both strains. They were distributed in areas of decreasing densities from an area centralis, located in the central upper retina to the retinal periphery. We have arbitrarilly grouped them in four classes of different densities: A class with 41 cells/mm2 and more, B class (from 31 to 40 c/mm2), C class (from 21 to 30 c/mm2) and D class (20 c/mm2 and less) and drawn isodensity lines. The comparison of cell mean density in each zone showed no major differences between the strains. Conversely, the comparison of the relative retinal surface occupied by each zone demonstrated a significant variation of C and D zones which appeared to be counterbalanced: the C zone being larger and the D zone smaller in pigmented quails compared with mutant quails. The possible involvement of such a TH-I cell distribution and consequently of the dopaminergic system in the development of glaucoma is discussed.

Dopaminergic interplexiform cells in the retina of pigmented and hypopigmented quails (Coturnix coturnix japonica).

Interplexiform cells (IPCs) have not been previously described as a component of the population of tyrosine hydroxylase (TH) immunoreactive (presumably dopaminergic) cells in the avian retina. In this study, carried out in both pigmented and imperfect albino mutant quails (Coturnix coturnix japonica), we initially describe TH immunoreactive cells in the inner nuclear layer whose internal dendritic arborization extends into strata 1, 3 and 4/5 of the inner plexiform layer. Then, we describe ascending processes arising from the somata or proximal dendrites of these cells. These sclerally directed processes (100-1,000 microns long) run across the inner nuclear layer to terminate within the outer plexiform layer, sometimes even reaching the outer nuclear layer. Hence, the cells bearing such processes correspond well with the definition of IPCs. The number of scleral processes is higher in mutant (48 +/- 19/retina) than in normal (12 +/- 10/retina) quails and are distributed throughout the retina except the area surrounding the pecten. Comparison of biochemical assays for dopamine in the two strains reveals a significantly higher dopamine content in the mutant quails which could be related to its increased number of dopaminergic IPC processes.