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Biomedical subjects

R Adler

Publications and source records attributed to R Adler.

At least 127 records · Page 7Linked to original sources
PubMed1989

Na+,K+-adenosine triphosphatase polarity in retinal photoreceptors: a role for cytoskeletal attachments.

We have used isolated embryonic photoreceptor cells as a model system with which to examine the mechanisms responsible for the development and maintenance of asymmetric Na+,K+-ATPase (ATPase) distribution. Photoreceptor precursors, which appear round and process free at culture onset, develop structural and molecular properties similar to those of photoreceptor cells in vivo. ATPase, recognized by an anti-ATPase antibody, is distributed over the entire surface of round photoreceptor precursors. As the cells develop, ATPase becomes progressively concentrated in the inner segment (where it is found in cells of the intact retina). This phenomenon occurs in cells developing in the absence of intercellular contacts. The development of ATPase polarity correlates with a decrease in the fraction of ATPase molecules that are mobile in the membrane (as determined by fluorescence photobleaching recovery), as well as with an increase in the fraction of ATPase that remains associated with the cells after detergent extraction. The magnitudes of the mobile ATPase fractions agree well with those of the detergent-extractable fractions in both the immature and developed photoreceptors. The distribution of alpha spectrin and ATPase-immunoreactive materials appeared qualitatively similar, and quantitative image analysis showed similar gradients of spectrin and Na+,K+-ATPase immunofluorescence along the long axis of elongated photoreceptors. Moreover, detergent extractability of alpha spectrin and the ATPase showed similar modifications in response to changes in pH or KCl concentration. ATPase detergent-extractable and mobile fractions were not changed in cultures treated with cytoskeletal inhibitors such as nocodazole. These data are consistent with a role for an asymmetrically distributed, spectrin-containing subcortical cytoskeleton in the preferential accumulation of Na+,K+-ATPase in the photoreceptor inner segment.

Animals↗
PubMed1989

Child behavior ratings: the influence of maternal characteristics and child temperament.

This study investigated the extent to which maternal characteristics, such as psychological health problems, marital adjustment and confidence in mother/wife roles, influenced how mothers rated the behavior of their first-born children (n = 100) on the Pre-school Behavior Questionnaire. Results showed that these characteristics were powerful predictors of behavior ratings. In contrast, the independent contribution of child temperament ratings was relatively small. It was concluded that child behavior problem and temperament measures may be confounded. Maternal ratings constitute a valuable source of information concerning parent-child interaction which deserve further investigation, especially of their influence on generally accepted measures of child behavioral adjustment.

Adaptation, Psychological↗
PubMed1989

Ganser syndrome in a 10 year old boy--an 8 year follow up.

A case of pseudodementia in a 10 year old boy was first reported in 1981. When reviewed 8 years after initial presentation, his symptoms were largely unchanged. The case continues to be complicated by claims for compensation and a successful appeal against the initial compensation judgment.

Amnesia↗
PubMed1989

Differential susceptibility of isolated mouse retinal neurons and photoreceptors to kainic acid toxicity. In vitro studies.

Dissociated cultures of mouse retinal neurons and photoreceptors in chemically defined medium were used to investigate the susceptibility of these cells to the neurotoxin kainic acid (KA). Cells isolated from the newborn mouse retina were initially insensitive to this toxin, and the cells that differentiated as rod photoreceptors retained this resistance throughout the culture period. However, amacrine neurons became increasingly sensitive to KA toxicity as they differentiated in culture; after the fifth day in vitro approximately 90% of these cells were killed by KA in a concentration- and time-dependent fashion. The neurons showed pronounced swelling within 10 min of treatment onset, and cell lysis and nuclear fragmentation were evident during the next few hours. KA-induced degeneration of these neurons was corroborated using the amacrine cell-specific monoclonal antibody HPC-1, as well as autoradiographic and biochemical determinations of the high affinity uptake for GABA. This inhibitory neurotransmitter was taken up by amacrine neurons but not by photoreceptor cells, and this uptake was completely abolished in KA-treated cultures. Similar results were obtained with the neuronal enzyme choline acetyltransferase. However, both photoreceptor survival and the expression of photoreceptor markers such as opsin and the retinoid-binding protein interphotoreceptor (IRBP) were similar in KA-treated and control cultures. Similarly, the high affinity uptake of glutamate, an excitatory amino acid which is predominantly taken up by photoreceptors, showed only modest changes in KA-treated cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Proteoglycan synthesis in flat cell-free cultures of chick embryo retinal neurons and photoreceptors.

Extracellular matrix and cell surface proteoglycans are thought to play important roles in neural development and regeneration. Central nervous system proteoglycans have been isolated and characterized from rat and sheep brain and from chick neural retina. An experimental advantage offered by the latter tissue is that it is avascular and can be isolated free of connective tissue and pigment epithelium. Therefore, proteoglycans synthesized by this tissue are derived exclusively from neural cells. However, it has not yet been determined whether neurons and photoreceptors contribute to proteoglycan synthesis or whether these molecules are largely glial in origin. In the present study we have addressed this question using cultures of chick neural retinal cells free of flat, glial-like cells. Proteoglycans synthesized by cultures of retinal neurons, photoreceptors, and undifferentiated, process-free round cells from 8-day embryonic chick neural retina were metabolically labeled in vitro using [35S]sulfate and [3H]glucosamine as precursors. Radiolabeled proteoglycans accumulated in the medium, and could also be extracted from the cell layer by sequential treatments with Triton X-100 and with guanidine HCl. The proteoglycans were isolated by ion-exchange chromatography, and characterized by gel filtration chromatography and by susceptibility to degradation by enzymatic and chemical treatments. Overall, heparan sulfate proteoglycans were the predominant type of proteoglycan synthesized in vitro by the cultured neural retinal cells at this developmental stage. The medium and the Triton extract contained different proportions of both chondroitin sulfate and heparan sulfate proteoglycans, while heparan sulfate was the only proteoglycan recovered from the guanidine extract. These studies demonstrate that heparan sulfate and chondroitin sulfate proteoglycans are actively synthesized by cultures of neural retinal cells free of flat, glial-like cells.

Animals↗
PubMed1988

Selective failure of long-term survival of isolated photoreceptors from both homozygous and heterozygous rd (retinal degeneration) mice.

Retinas from homozygous rdle/rdle and heterozygous rdle/++ C57BL/6J mice were dissected and dissociated on postnatal day 2, when they are still essentially indistinguishable. The resulting cell suspensions were seeded on highly adhesive substrata, to which the cells attach as individual units, and grown in vitro for 2 weeks in serum-free, chemically defined media. The behavior of neurons and photoreceptors in vitro was investigated with several techniques; essentially no differences were found between rdle/rdle and rdle/++ cells. Three distinctive cell types could be recognized in cultures of both genotypes towards the end of the first week in vitro: process-free cells, multipolar neurons and rod photoreceptors. There were similarities between rdle/rdle and rdle/++ cultures in the number and morphology of photoreceptor cells, to include the presence of a cilium and a short neurite terminating in a spherule-like body. Moreover, in cultures of both genotypes, only photoreceptors showed opsin immunoreactivity and the antigen recognized by the rod-specific monoclonal antibody RET-P1. Biochemical and autoradiographic studies demonstrated that rdle/rdle and rdle/++ cells also showed similar uptakes of the putative amino acid neurotransmitters glutamate and aspartate (associated with most of the photoreceptors and only some neurons), and gamma-aminobutyric acid (associated with neurons but absent in photoreceptors). Thus, according to several parameters, the properties shown by photoreceptor cells were similar in rdle/rdle and rdle/++ cultures during the first week in vitro. Massive photoreceptor cell death was observed in both genotypes during the second week in vitro, coinciding with the time when photoreceptor degeneration occurs in vivo in rd/rd, but not in rd/+ retinas. Photoreceptor death in culture appeared to be specific, since approx. 80% of the non-photoreceptor neurons survived normally during the period when photoreceptor degeneration took place. Several reports from the literature suggest that the period around postnatal days 8-10 represents a critical stage for rd/rd photoreceptors, since they survive until this time but degenerate thereafter. Genetically normal photoreceptors apparently undergo a comparable crisis during maintenance in primary culture, suggesting the involvement of cell-cell contacts and/or retina-derived environmental signals in the survival or rod visual cells.

Amino Acids↗
PubMed1988

Induction of antioxidant enzyme activities by a phenylurea derivative, EDU.

Oxygen free radicals have the potential to mediate cell injury. Defenses against such radicals include the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX). The purposes of this study were (1) to develop an in vitro model using human cells in which to investigate a potential pharmacologic agent as an inducer of these antioxidant enzymes; (2) to investigate the phenylurea derivative N-[2-(2-oxo-1-imidazolindinyl)ethyl]-N-phenylurea (EDU) in this model with paraquat (PQ) serving as the positive control; and (3) to determine if induction of the antioxidant enzymes by EDU occurs in vivo. Human gingival fibroblasts (Gin-1) were used as the target cell in vitro; PQ and EDU, an inducer of SOD and CAT activities in plants, were evaluated as antioxidant enzyme inducers. Total SOD activity in Gin-1 cells increased 2-fold (p less than 0.05) in the presence of 1.0 mM PQ for 18-48 hr compared with untreated controls. Gin-1 cells incubated with 0.25-2.0 mM PQ for 24 hr had significantly increased total SOD (1.5 to 2.0-fold; p less than 0.05). CAT activity increased with 1.0 and 2.0 mM PQ (p less than 0.05). In the presence of PQ, GSH-PX activity decreased (p less than 0.05) in a concentration-dependent manner, indicating inactivation of this enzyme. No toxicity, indicated by lactate dehydrogenase released into the incubation medium, was noted at PQ concentrations below 5.0 mM. In the presence of 0.125-2.0 mM EDU, total SOD activity in Gin-1 cells significantly increased (1.5 to 2.0-fold; p less than 0.05). CAT activity significantly increased in a dose-dependent manner (p less than 0.05), while GSH-PX activity remained constant following exposure to 0.125-2.0 mM EDU. Intraperitoneal administration of EDU to rats twice a day for 2 days at 100 mg/kg induced SOD activity in heart, liver, and lung compared to controls (p less than 0.05). CAT activity increased in the liver 56% and in the lung 36% (p less than 0.05). GSH-PX activity remained constant. Our findings indicate that Gin-1 cells are a useful model in which to study inducers of antioxidant enzymes in vitro and that the phenylurea compound EDU induces SOD and CAT activities both in vitro and in vivo.

Antioxidants↗
PubMed1988

Doxorubicin conjugates of monoclonal antibodies to hepatoma-associated antigens.

A panel of six murine monoclonal antibodies against hepatocellular carcinoma-associated antigens, reactive with PLC/PRF/5 human hepatoma cells, was conjugated to Adriamycin (doxorubicin) via a dextran bridge. This library of antibodies includes three monoclonal antibodies against hepatitis B virus surface antigen, one anti-alpha-fetoprotein, and two other IgG2a antibodies against PLC/PRF/5 hepatoma-associated antigens. The use of dextran for conjugation of Adriamycin to antibodies enabled a 5- to 10-fold amplification of the number of drug molecules linked to antibody. Conjugation of Adriamycin to dextran caused an occasional reduction in the pharmacologic activity of dextran-Adriamycin in [3H]thymidine incorporation assays in hepatoma cells as compared to nonconjugated Adriamycin. This loss of anticellular activity was partially compensated for by conjugation of specific antibodies to the dextran-Adriamycin conjugate. Conjugated compounds completely retained their binding activity to purified hepatitis B virus surface antigen and alpha-fetoprotein fixed to a solid matrix as compared to binding of homologous nonconjugated antibodies. However, some reduction of the binding activity to intact hepatoma cells was observed in three of six conjugates. Binding activity to hepatoma cells and, as a consequence, suppression of tumor cell DNA synthesis by the various conjugates was enhanced as compared to the same effect in treated colorectal carcinoma cells that do not express the relevant hepatoma-associated proteins. Furthermore, two conjugates containing nonspecific antibodies did not bind to hepatoma cells and caused minimal suppression of DNA synthesis. These results suggest that this panel of monoclonal antibody-dextran-Adriamycin conjugates was effective in suppression of PLC/PRF/5 cell growth in vitro.

Animals↗
PubMed1988

Development of neonatal mouse retinal neurons and photoreceptors in low density cell culture.

We describe here a culture method which allows the growth of dissociated mouse retinal neurons and photoreceptors in chemically defined medium. Neural retinas from 2-day-old C57/BL mice were dissected from other ocular tissues, including the pigment epithelium, and dissociated into a cell suspension after brief trypsination. Most cells attached as single, unaggregated units to substrata pretreated with polyornithine and the neurite-promoting factor (PNPF). The cells were cultured in serum-free, high pyruvate Dulbecco's modified Eagle's medium containing chemically defined supplements. Under these conditions, onset of cell process development was rapid, giving rise to extensive neurite networks. Three morphologically distinct cell types were apparent during the first week in vitro. Some cells retained a circular outline and failed to produce processes, while 50-60% of the cells developed as multipolar neurons showing a large cell body and several neurites. Approximately 90% of these cells reacted with an amacrine cell-specific monoclonal antibody. Some 30% of the cultured cells expressed phenotypic properties characteristic of rod photoreceptors, including a small cell body, an apical cilium, a short neurite with a spherule-like terminal body, and immunoreactivity with antibodies against opsin as well as a rod cell-specific monoclonal antibody. No further signs of outer segment differentiation were observed in these cells. Non-neuronal "flat" cells, which represented less than 0.5% of the total cell number, reacted with an antibody against the glial fibrillary acidic protein. The number of neurons and photoreceptors remained relatively stable during the first 4-7 days in vitro. During the second week in culture, however, there was specific degeneration of greater than 90% of the photoreceptor cells, while less than 20% of the multipolar neurons were similarly affected. Consequently, in addition to providing a system for studying the differentiation of retinal neurons and photoreceptors, the specific degeneration of photoreceptors in these mouse retinal cell cultures makes this system ideal for investigating factors influencing photoreceptor survival.

Animals↗
PubMed1987

Nature and nurture in the differentiation of retinal photoreceptors and neurons.

This article reviews recent studies using a novel experimental system in which undifferentiated precursor cells from the 8-day chick embryo retina are grown in low density, clump-free, dissociated cell culture. The cultures initially consist of a morphologically homogeneous population of isolated process-free, round cells. Analysis of the cultures by phase contrast light microscopy, scanning and transmission electron microscopy, immunocytochemistry and autoradiography, shows that during the first week in vitro some precursor cells acquire a well differentiated photoreceptor phenotype, while others develop as neurons. Given that these divergent differentiation pathways are followed by cells developing in a homogeneous microenvironment in the absence of intercellular contacts, the evidence suggests that precursor cells present in the 8-day chick embryo retina are already preprogrammed to undergo an extensive series of chemical and structural modifications necessary to differentiate as either neurons or photoreceptors.

Animals↗
PubMed1987

Differential sensitivity of cultured retinal neurons and photoreceptors to herpes simplex infection.

Infection of the retina with herpes simplex virus type 1 (HSV-1) causes devastating lesions usually leading to blindness. However, the interactions between individual retinal cell types and this virus have not been well characterized, probably because of limitations posed by the complexity of the intact retina. We have now approached this problem through the use of separate, purified populations of isolated chick embryo retinal neurons and photoreceptor cells, of glial cells, and of pigmented epithelial cells. This manuscript deals with the initial part of these studies, aimed at determining the susceptibility of different retinal types to HSV-1 infection. The different cultures were exposed to HSV-1 for 3-48 hr, and cell infection was evaluated by immunocytochemical detection of viral antigens or by autoradiographic study of viral DNA replication. Practically 100% of the retinal glial cells and pigmented epithelial cells appeared susceptible to HSV-1 infection. On the other hand, as many as 70% of the neurons present in glia-free, pigment epithelium-free cultures, also appeared infected after a 24-hr exposure to the virus. Neuronal susceptibility to HSV-1 was already present in early (2-day) cultures, was time- and concentration-dependent, and led to neuronal degeneration after 24-48 hr. Neuronal infection was also corroborated by the detection of viral particles by transmission electron microscopy. Photoreceptor cells were consistently and selectively resistant to HSV-1 infection at all the concentrations and time points investigated. Both immunocytochemical and autoradiographic studies showed similar results. Photoreceptor resistance to HSV-1 appears to be selective, since they could be readily infected with RNA viruses such as vesicular stomatitis virus and influenza virus. These cell culture preparations offer an attractive system for the investigation of cellular mechanisms involved in the differential susceptibility of retinal cells to viral infection. Moreover, they could also help in the screening of treatments potentially capable of preventing and (or) curing HSV-induced retinal infection.

Animals↗
PubMed1987

Selective destruction of photoreceptor cells by anti-opsin antibodies.

This article reports that cultured photoreceptor cells are selectively vulnerable to complement-mediated lysis by antiopsin antisera. The study has been carried out using a culture system that permits the growth of embryonic retinal neurons and photoreceptor cells in the absence of glial, pigment epithelial, connective, or endothelial cell contamination. Exposure of these cultures to an antiopsin antiserum in the presence of complement results in the lysis of 85% of the photoreceptor cells, without extensive loss of neurons. Photoreceptor lysis by the antibody is dose-dependent and specific, as demonstrated by the ability to block photoreceptor lysis by preincubation of the antiopsin antiserum with purified rhodopsin. Photoreceptor sensitivity to lysis by these antibodies develops in vitro in parallel with the appearance of immunocytochemically detectable opsin. Multipolar neurons (which do not contain opsin) are not affected by the treatment, as shown by microscopic analysis and determination of neuronal "markers" such as choline acetyltransferase and GABA high affinity uptake. These results show that antiopsin antibodies can cause selective photoreceptor degeneration. Moreover, this in vitro system appears useful as a bioassay to test a variety of possible mechanisms of retinal cell destruction which may be important in vivo. The results suggest also a mechanism for the generation of enriched retinal neuronal cultures by selective lysis of photoreceptor cells.

Animals↗
PubMed1986

Developmental predetermination of the structural and molecular polarization of photoreceptor cells.

Through mechanisms still unknown, the apparently homogeneous neuroepithelium of the embryonic optic cup differentiates into such divergent cell types as photoreceptors, glia, and various subsets of neurons. Questions that still remain unanswered in this field include the timing and mechanism of action of the "instructive" events directing each neuroepithelial cell to undergo the sequence of phenotypic changes necessary to develop into a specific retinal cell type. This laboratory is investigating some of these questions using cultures in which dissociated neural retina cells, obtained before the onset of overt photoreceptor differentiation, develop at low density in the absence of glia and pigment epithelium. The cultures initially are a morphologically homogeneous population of process-free, round cells. Some cells retain this morphology throughout the first week in vitro, while others develop either as photoreceptors or as multipolar neurons. Photoreceptors elongate and become very asymmetric as they do in vivo, with characteristic compartments orderly arranged along their longitudinal axis (an outer segment-like process, inner segment, cell body, and a characteristically short, single neurite). Cell polarization can also be observed in the distribution of opsin immunoreactive materials and some cytoskeletal elements. Thus, certain precursor cells present in the embryonic retina seem to be programmed to differentiate into photoreceptors even when developing in the absence of contacts with other retinal cells. However, interactions with other constituents of the retina/pigment epithelium complex are probably necessary to ensure final photoreceptor maturation, including further growth of the opsin-rich outer segment process.

Animals↗