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P Madtes

Publications and source records attributed to P Madtes.

14 recordsLinked to original sources

Postnatal development of 3H-GABA-accumulating cells in rabbit retina.

Light and electron microscopic autoradiography demonstrates that 3H-GABA is accumulated by horizontal cells in neonatal rabbit retina but not in the adult. A specific population of horizontal cells appears to be mature at birth and they avidly accumulate 3H-GABA during a 15-minute incubation period in vitro. Uptake into horizontal cells is not observed after the fifth postnatal day; 3H-GABA-accumulating horizontal cell bodies and their processes are the first identifiable components that clearly mark the future location of the outer plexiform layer at birth and as such, may be considered pioneering elements. Our observations raise the interesting possibility that the pioneering horizontal cell may provide structural and/or chemical factors necessary for the subsequent development of the outer plexiform layer of the retina. Labeling patterns of other retinal cells also show varying degrees of change during development. A population of amacrine cells accumulate 3H-GABA at birth. These cells show little change in their morphological or 3H-GABA uptake properties from birth to adulthood. Müller cells show weak accumulation of 3H-GABA at birth. Subsequent to this time, labeling of Müller cells is significantly more robust, resulting in Müller cell domination of retinal autoradiographic patterns in more mature retinas. Every cell body in the ganglion cell layer accumulates 3H-GABA at birth. The number of labeled cells declines during postnatal development, resulting in a very limited adult population. We conclude that the ability of retinal cells to accumulate 3H-GABA does not remain constant during postnatal development; rather each cell population displays a unique maturation sequence that results in a dramatic developmental shift in the number and types of GABA-accumulating cells present in the retina.

Age Factors

GABA receptor binding site "induction" in rabbit retina after nipecotic acid treatment: changes during postnatal development.

The development of the GABA system in the rabbit retina was studied. The number of high- and low-affinity GABA receptor binding sites increased in a sigmoidal manner, with the curve for the low-affinity sites lagging 2-3 days behind that for the high-affinity sites. The KD for both high- (17.5 nM) and low-affinity (138.0 nM) sites remained constant during development. Treatment of isolated eyecups with the uptake blocker nipecotic acid resulted in an increase in the Bmax for high-affinity sites in developing tissue with the maximum sensitivity around eye opening; mature tissue exhibited a decrease in Bmax. In contrast, a gradual decrease in sensitivity to stimulation of the low-affinity sites occurred. These data indicate that the "trophic" action of GABA is limited to the time when the tissue is developing.

Age Factors

Metabolism of [3H]nipecotic acid in the rabbit retina.

Nipecotic acid has been demonstrated to block the gamma-aminobutyric acid transport systems. Previous studies have shown that the uptake system is the first transmitter-specific parameter to appear during the development of the rabbit retina. Use of these observations has been made to study the influence on the development of gamma-aminobutyric acid receptors of altering the uptake mechanism by treating newborn pups with nipecotic acid to block GABA transport. The present study of the in vivo metabolism of [3H]nipecotic acid in the CNS measured the changes in the levels of [3H]nipecotic acid in both adult and newborn rabbit retinas after injection of the label into the vitreal chamber. It was found that the effective half-life of [3H]nipecotic acid in the vitreous is about 5 h for adult tissue and 3 h for newborn. In contrast, all retinal fractions retained the label longer, the effective half-lives being about 60 h (adult) and 45 h (newborn). Further, no labeled metabolites of nipecotic acid were detected in either adult or newborn tissue. This study gives evidence that the degradation of nipecotic acid in nervous tissue is minimal and suggests that, although the rate of clearance is faster in neonates, the fate of nipecotic acid in vivo may be similar in both adult and newborn tissues.

Age Factors

Neurotransmitter systems in morphologically undifferentiated human Y-79 retinoblastoma cells: studies of GABAergic, glycinergic, and beta-adrenergic systems.

Elements of three neurotransmitter systems were investigated in morphologically undifferentiated human Y-79 retinoblastoma cells in suspension culture. Specific gamma-aminobutyric acid (GABA) uptake, GABA binding, and glycine binding were absent from these cells, although the cells had been shown to exhibit an active uptake and release of [3H]glycine. Binding and competition studies using both alpha- and beta-adrenergic ligands indicated the presence of a beta-adrenergic receptor. This finding was confirmed by treatment of the cells with beta-agonists in competition with a beta-antagonist and with an alpha-antagonist; the level of cyclic AMP was competitively stimulated. Therefore, human Y-79 cells in suspension culture contain beta-adrenergic receptors, and not glycinergic or GABAergic ones. Thus, the Y-79 cells may be of use in studying the factors involved in developmental regulation of neurotransmitter function.

Adrenergic beta-Agonists

Chloride ions preferentially mask high-affinity GABA binding sites.

Chloride ions (Cl-) act to regulate the appearance of gamma-aminobutyric acid (GABA) binding sites in synaptic membranes from bovine retinas. Scatchard analysis indicates that the presence of Cl- results in a preferential decrease (85%) in the number of high-affinity binding sites, with no statistically significant change in the affinity (KD) of either the high- or low-affinity sites. The finding that the appearance of this binding site is altered by the presence of Cl- may indicate that Cl- acts, either directly or indirectly, to regulate the appearance, and hence function, of the high-affinity GABA receptor.

Animals

GABA as a trophic factor during development.

The process of synaptogenesis has been studied by many investigators to determine the factors which regulate synapse formation. We have used neonatal rabbit retina to investigate the role of the gamma-aminobutyric acid (GABA) neurotransmitter system during development. By utilizing an in vitro incubation treatment of isolated eyecups we found that treatment with nipecotic acid, a GABA uptake blocker, resulted in a 4-fold increase in the amount of specific 3H-muscimol binding. In addition, incubation of the tissue in the presence of the GABA agonists muscimol, 4,5,6,7-tetrahydroisoxazolo [5,4-c]pyridine-3-ol (THIP), or GABA itself led to similar increases in specific 3H-muscimol binding. The findings support the conclusion from previous studies that the induction of GABA receptors observed after in vivo treatment of 1-day-old rabbits with nipecotic acid resulted from an increase in the extracellular concentration of GABA. A possible role for GABA in the regulation of GABAergic synapse formation is presented in this report.

Animals

Intraocular injections of nipecotic acid produce a preferential block of neuronal 3H-GABA accumulation in adult rabbit retina.

A procedure by which the activity of the retinal GABA uptake system can be manipulated in vivo has been developed. Intraocular injections of nipecotic acid, a proported GABA uptake blocker, were administered to adult rabbits every 48 hours for a two-week period. No behavioral or systemic changes were observed. Injections were well-tolerated with less than 10% loss of the tissue caused by physical damage or injection. Biochemical analyses demonstrated a dose-dependent inhibition of 14C-GABA uptake into retinal tissue. No effect on uptake was observed for saline-treated tissue. Autoradiographic analyses showed that in vivo treatment with nipecotic acid preferentially blocked accumulation of 3H-GABA into the amacrine cell bodies and processes in the inner plexiform layer. This treatment may be especially useful in assessing the functional significance of GABA transport in vivo.

Animals

[3H]GABA binding in developing rabbit retina.

We have studied the developmental sequence of the GABA system in the rabbit retina using an in vitro binding assay to monitor developmental changes in the post-synaptic receptor. A variety of tissue treatments including perchlorate and Triton X-100 were employed to optimize binding and remove endogenous factors which inhibit binding. Pre-treatment of the tissue with 0.05% Triton X-100 revealed high affinity binding for [3H]GABA which increased in a sigmoidal fashion with the post-natal age of the animal. A constant level of binding, at about 16% of adult levels, was noted until day 8, at which time a rapid increase occurred. At 16 days post-natal, the amount of specific binding reached a plateau near adult levels. Kinetic analysis of the GABA receptor showed an increase in the number of receptors (Bmax) with little or no change in the apparent affinity (KD). Our results suggest that the onset of post-synaptic receptor activity is delayed approximately 1 to 2 days, relative to the pre-synaptic components, and the period of rapid increase in GABA receptor binding coincides with the period of maximum increase in retinal synaptic density.

Aging

'Binding' of glycine and gamma-aminobutyric acid to synaptosomal fractions of 6 regions of the feline brain; effects of strychnine.

GABA (6 X 10(-6) M) binding to synaptosome-enriched fractions of cat CNS exhibited a clear rostro-caudal gradient, whereas glycine (6 X 10(-6) M) binding was greatest to particles of cerebellar cortex, and this was followed by medulla approximately equal to caudate nucleus larger than or equal to cerebral cortex larger than or equal to pons larger than corona radiata. Strychnine-SO4 (10(-3) or 10(-4) M) inhibited the binding of GABA and glycine in all brain regions studied; at 10(-5) M this drug inhibited the binding of both GABA and glycine only to particles of the cerebral cortex.

Aminobutyrates

'Binding' of glutamate and asparatate to synaptosomal fractions of six regions of the feline brain.

The order of potency of 'binding' of both glutamate and aspartate to synaptosomal fractions of brain regions was: cerebellar cortex greater than caudate nucleus greater than or equal to cerebral cortex greater than medulla approximately pons greater than corona radiata. Glutamate was bound to a greater extent than aspartate to particles of all regions studied, except for cerebral cortex.

Animals