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A J Adler

Publications and source records attributed to A J Adler.

At least 37 records · Page 2Linked to original sources
PubMed1992

Androgen-specific gene activation via a consensus glucocorticoid response element is determined by interaction with nonreceptor factors.

A fundamental issue in steroid hormone regulation is the question of how specific transcription is attained in vivo when several receptors can bind the same DNA sequence in vitro. We report an enhancer of the mouse sex-limited protein (Slp) gene that, unlike previously characterized enhancers, is activated by androgens but not by glucocorticoids or progestins. Potent androgen induction requires both a consensus glucocorticoid (hormone) response element and auxiliary elements also present within a 120-base-pair DNA fragment. Cotransfection assays with wild-type and mutant receptors reveal that glucocorticoid receptor can bind, but not transactivate from, the hormone response element within the enhancer. The positive effect of androgen and the null effect of glucocorticoid appear to require the amino-terminal domains of the respective receptors. Thus, exclusive transcriptional response to androgens, and lack of response to glucocorticoids, derives from factor interactions that are determined by the context of the receptor binding site rather than by its distinct sequence.

Androgens↗
PubMed1992

Synthesis of retinoic acid from retinol by cultured rabbit Müller cells.

Previous observations have shown that Müller glial cells of the vertebrate retina contain cellular retinoid-binding proteins, that the retina contains retinoic acid, and that cellular retinoic acid-binding protein is present in amacrine neurons (and, in some species, Müller cells) within the retina. These findings led to the suggestion that Müller cells may synthesize retinoic acid and release it for use by other retinal cells. To test this possibility, we cultured Müller cells from adult rabbit retinas, incubated the cultures with radioactive retinol, and identified and quantified the resultant radioactive retinoids by HPLC. Retinaldehyde was rapidly synthesized from retinol, reaching a plateau of 1-2 pmol mg-1 cell protein by 30 min. Retinoic acid initially accumulated more slowly, but by 30 min constituted most of the synthesized retinoid. While the retinaldehyde remained within the cells, retinoic acid was rapidly released into the medium; extracellular retinoic acid exceeded the intracellular amount after 30 min of incubation. Smaller amounts of retinyl esters were also synthesized and retained by the cells. These results are consistent with the suggestion that Müller glia are a source of retinoic acid in the retina. The synthesis of retinoic acid by these cells, and the presence of retinal neurons that contain cellular retinoic acid-binding protein, raise the possibility that retinoic acid plays a role in the retina, although this role is not presently known. Furthermore, these results may have implications for other parts of the adult nervous system. Adult brain contains retinol- and retinoic acid-binding proteins, and, therefore, may also be a site of retinoic acid metabolism. Because of the relatively simple cellular organization of the retina and its demonstrated capacity to synthesize retinoic acid, the retina may be a system of choice for further studies of the synthesis and function of retinoic acid in adult neural tissue.

Animals↗
PubMed1992

Distribution of glucose and lactate in the interphotoreceptor matrix.

The photoreceptor cells of the retina derive their nourishment from the choroidal blood supply behind the retina. If the extracellular interphotoreceptor matrix (IPM) mediates this retinal nutrition, then gradients in concentrations of nutrients such as glucose and waste products such as lactic acid would be expected across the IPM, from the neural retina to the retinal pigment epithelium (RPE); this hypothesis is examined here. Gentle methods were employed to prepare IPM from the retinal surface (IPM-R) and apical RPE surface (IPM-P) of bovine eyes, and contamination from adjacent tissues was very low. The actual intrinsic volumes of the IPM-R (180 microliters) and IPM-P (87 microliters) compartments were estimated from dilution of trypan blue dye added to the wash buffer used for sample preparation; these measurements were required for calculation of concentrations. In the IPM-R samples glucose was virtually undetectable and the lactate concentration was very high (13 mM). Near the RPE surface (IPM-P) both compounds were present at levels (glucose 0.9 mM, lactate 3.8 mM) more comparable to those in blood serum. Similar results were obtained with fresh rabbit eyes. This difference in distribution (nutrient more concentrated near RPE, waste product near retina) indicates gradients consistent with the utilization of the IPM as a pathway for outer-retinal nutrition.

Animals↗
PubMed1991

Requirement of insulin or IGF-1 for the maintenance of retinyl ester synthetase activity by cultured retinal pigment epithelial cells.

Previous work from these laboratories showed that the retention of retinyl ester synthetase activity by cultured human retinal pigment epithelium is up to tenfold greater with PM medium (Medium 199 plus insulin, other added defined components, 1% serum and 1% retina extract) than with conventional culture media. The present work shows that insulin is the component of PM medium required for maintenance of ester synthetase activity and that insulin-like growth factor type 1 (IGF-1) also is effective at maintaining ester synthesis. In addition, insulin can maintain ester synthetase activity in cultured rat RPE. Preliminary dose-response measurements provide additional support for these findings and strongly suggest that both insulin and IGF-1 are maximally effective at physiological concentrations (1-10 ng ml-1).

Acyltransferases↗
PubMed1991

Effect of light on endogenous ligands carried by interphotoreceptor retinoid-binding protein.

Interphotoreceptor retinoid-binding protein (IRBP) is a vitamin A carrier present only in the extracellular material lying between the neural retina and the retinal pigment epithelium of vertebrate eyes. The amount of retinol bound endogenously by IRBP in this interphotoreceptor space is known to increase upon illumination. This finding led to the hypothesis that IRBP may act as a shuttle for vitamin A during the visual cycle that regenerates rhodopsin. In the present work, we separated IRBP from other retinoid-binding proteins in bovine interphotoreceptor matrix preparations by means of size-exclusion chromatography. IRBP's endogenous ligands were retained during this procedure and were then extracted into hexane and analysed by normal-phase HPCL. We found that IRBP carries, in a light-dependent manner, all the retinoid isomers involved in the visual cycle. For dark-adapted eyes the amounts of bound ligands are (in nmol per eye) 0.09 all-trans retinol, 0.11 11-cis retinol, 0.04 all-trans retinal, 0.16 11-cis retinal, and 0.07 retinyl esters. For light-adapted eyes the amount of all-trans retinol was found to increase by a factor of five, and that of 11-cis retinal to decrease by a factor of four. (These eyes contain 3.1 nmol of IRBP, which does not change in amount with lighting conditions). Thus, the major endogenous ligand of IRBP is 11-cis retinaldehyde in the dark and all-trans retinol in the light. The data are consistent with a role for IRBP as a non-selective scavenger and stabilizer of retinoids released from photoreceptors and pigment epithelial cells. However, it cannot be concluded from these data that IRBP is involved in directed transport of retinoids across the interphotoreceptor matrix, since there is no evidence for the appropriate spatial gradients in the ligands bound to the protein.

Adaptation, Ocular↗
PubMed1991

Multiple components of a complex androgen-dependent enhancer.

Sex-limited protein (Slp) is expressed in adult male mice. A 160-basepair fragment 2 kilobases upstream of the gene serves as an androgen-dependent enhancer of chloramphenicol acetyltransferase expression in transient transfection assays in cells with endogenous or cotransfected androgen receptor. One element that is necessary, but not sufficient, for induction is a consensus glucocorticoid (or hormone) response element (HRE). This element binds to the mouse androgen receptor in vitro, but with apparent weak affinity. Induction by the HRE is greatly augmented by an accessory sequence within the 160 basepairs, suggesting that cooperative interactions confer strong response to androgen. Additional elements within the enhancer modulate induction, positively or negatively, and exhibit cell-specific behavior. Of particular interest are two degenerate HREs that are adjacent to the consensus sequence; they show no independent activity, but are functionally significant in conjunction with other elements. The complexity of this enhancer may reflect biological mechanisms that ensure specificity of hormonal response and allow gene expression to respond to changes in hormone concentration.

Animals↗
PubMed1991

Effect of aluminum on calcium binding to bovine intestinal calcium-binding protein.

Aluminum is known to inhibit vitamin D-dependent intestinal absorption of calcium. To investigate possible mechanisms of action, the effect of aluminum on the binding of calcium to bovine intestinal calcium-binding protein (CaBP) was studied by means of equilibrium dialysis. The results of these studies indicate that aluminum in concentrations up to 80 microM and up to 80 times the ambient calcium concentration did not significantly alter the affinity of CaBP for calcium (Kd = 0.17 microM) nor its molar capacity (2 mol/mol CaBP).

Aluminum↗
PubMed1991

Biological control of primate macular pigment. Biochemical and densitometric studies.

The amounts of zeaxanthin (Z) and lutein (L), the carotenoids constituting the primate macular pigment, were measured in the central retinas of monkeys (Saimiri sciureus and Macaca fascicularis). Two independent methods--reverse-phase high-performance liquid chromatography (HPLC) and microdensitometry--were used for analysis of the same set of retinas. Most of the measurements were made on retinas that had been fixed by glutaraldehyde-paraformaldehyde perfusion of the animal. Control experiments showed that this fixation did not interfere with the quantitative extraction and analysis of the carotenoids. The amount of macular pigment calculated from microdensitometry of the foveal region was proportional to the amount of pigment assayed by HPLC of the same retinal area, demonstrating that either method can be used reliably to rank the carotenoid content of aldehyde-fixed foveas. The optical density of pigment in the axial direction through the retina was higher than would be predicted if the pigment were randomly oriented. This is consistent with the idea that the nonrandom orientation of the dichroic macular pigment molecules found in previous studies contributes to increased optical filtering of the retinal image. Comparisons of the amounts of Z and L between the left and right eyes of the same monkey, within 1 mm of the foveal center, always showed excellent agreement (averaging a 5% difference for Z and 11% difference for L), whereas differences among individual monkeys were very large (up to fourfold for Z). These results indicate that the uptake and assimilation of the macular carotenoids are biologically regulated by selective mechanisms in primate retinas.

Animals↗
PubMed1991

Distribution of individual macular pigment carotenoids in central retina of macaque and squirrel monkeys.

The spatial distribution of lutein (L) and zeaxanthin (Z), the structural isomers composing the macular pigment, was studied in the retinas of macaque monkeys (Macaca fascicularis) and squirrel monkeys (Saimiri sciureus). Spatial profiles of macular pigment optical density were obtained from retinal whole mounts. Then concentric annuli were microdissected from the fovea and adjacent regions of the same retinas. Each retinal segment was analyzed for carotenoids by high-performance liquid chromatography. Both L and Z reached their highest concentrations at the center of the fovea and declined monotonically with eccentricity for both primate species. This is inconsistent with a preferential association of L with rods. Macaque monkeys have a consistent pattern of more Z than L at the foveal center, like humans. Z declines more rapidly than L with eccentricity, so that L becomes dominant in the periphery. Squirrel monkeys (all male) showed striking individual differences. Some had more Z than L at the foveal center like macaques, but four of six had the reverse pattern, with more L than Z throughout the central retina. Individual differences among squirrel monkeys may be linked to their color vision polymorphisms. This suggests that a particular Z/L ratio in primate retinas may be associated with a specific cone phenotype, just as particular carotenoids are associated with specific cone types in vertebrates with cone oil droplets.

Animals↗
PubMed1990

Neural cell adhesion molecule (NCAM) in adult vertebrate retinas: tissue localization and evidence against its role in retina-pigment epithelium adhesion.

The presence of neural cell adhesion molecule (NCAM) was examined in the neural retina, interphotoreceptor matrix (IPM), and retinal pigment epithelium (RPE) of adult bovine and frog eyes. Using polyclonal antibodies raised against adult isoforms of NCAM. Western blot analyses revealed the presence of NCAM in the neural retina, but not in the IPM or RPE of these species. As a control, Western blot analysis was used to demonstrate the presence of interphotoreceptor retinoid-binding protein (IRBP) in the IPM preparations. NCAM immunoreactivity was detected by light microscopic immunocytochemistry primarily in the plexiform layers and nerve fibre layer of the frog retina. Minor immunoreactivity was also detected in the inner and outer nuclear layers, but there was no detectable NCAM immunoreactivity in the IPM, outer segments, or RPE. These results indicate that NCAM is not a likely participant in the process of retina-RPE adhesion in the adult eye.

Animals↗
PubMed1990

Inhibition of superoxide dismutase activity by silicon.

Elevated silicon levels have been found in the serum of uremic patients, in the brain of patients with senile dementia and in neuroglial tangles of Alzheimer patients. The effect of silicon on superoxide dismutase was studied in vitro, since excessive superoxide production occurs in renal failure, in inflammatory conditions and in the aging process. Silicon in concentrations similar to those found in serum of uremic patients inhibits superoxide dismutase activity. The degree of inhibition is directly proportional to silicon levels. Depression of superoxide dismutase by Si is likely to result in a decrease in oxygen free radical destruction and thus an increase in excessive local availability of oxygen free radicals. The increased silicon levels in brain, kidney, lung and RBC which are especially sensitive to oxygenation damage may contribute to a variety of important clinical complications, by means of excess oxygen free radicals.

Animals↗
PubMed1989

Pentafluorobenzoic anhydride as a derivatizing agent for alcohols and hydroxy fatty acid methyl esters detected by electron capture in gas chromatography.

Pentafluorobenzoate derivatives of primary and secondary alcohols have been prepared using pentafluorobenzoic anhydride. The gas chromatographic properties of the homologous series of the methyl esters of 2-hydroxycarboxylic acids from C12 to C26 have been studied on the high-temperature stationary phase Poly-S 179. The suitability of determining trace amounts of these compounds using gas chromatography with an electron-capture detector and with electron-capture negative-ion chemical-ionization mass spectrometry is discussed.

Anhydrides↗
PubMed1989

Selective presence of acid hydrolases in the interphotoreceptor matrix.

Adler and Martin (1983, Curr. Eye Res. 2, 359-66) found cathepsin D to be present in crude preparations of bovine interphotoreceptor matrix (IPM). The purpose of the present study was to determine, by investigating several acid hydrolases in purer IPM samples, whether hydrolytic enzymes abundant in RPE lysosomes were present also as normal components of the IPM. IPM was prepared from bovine eyes by the introduction of a small bleb of buffer between the neural retina and the RPE. These IPM samples were free from significant contamination by surrounding tissues; they contained IRBP as their only major protein, and had negligible amounts of lactate dehydrogenase and ROS-specific proteins. Most acid hydrolases were assayed fluorometrically by measuring the 4-methylumbelliferone released upon hydrolysis of appropriate derivatives; the substrate for cathepsin was hemoglobin. The amounts of the enzymes found in the IPM were far from uniform and could not be correlated with enzyme activities in either RPE or retina homogenates. The hydrolases in the IPM varied in amount from beta-galactosidase (28% of the RPE level), through N-acetyl-beta-glucosaminidase (20%), alpha-fucosidase (15%), beta-glucuronidase (12%), alpha-glucosidase (8%), cathepsin D (7%), alpha-mannosidase (7%), down to beta-glucosidase, acid phosphatase, and acid lipase (trace amounts, less than 1%). These results agree with the relative amounts of enzymes found by Wilcox (1987) to be secreted into the medium by cultured human RPE cells. Furthermore, the rank order of hydrolases in the IPM is the same as that for hydrolases secreted (but not recaptured) by human fibroblasts in I-cell disease. The conclusion from these correlations is that lysosomal enzymes are probably secreted, as a normal process, by the RPE into the IPM, where they may have a role in digesting shed outer segments and in catabolizing IPM components.

Acid Phosphatase↗