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G W Goldstein

Publications and source records attributed to G W Goldstein.

At least 19 recordsLinked to original sources
PubMed1992

Inhibition of astroglia-induced endothelial differentiation by inorganic lead: a role for protein kinase C.

Microvascular endothelial function in developing brain is particularly sensitive to lead toxicity, and it has been hypothesized that this results from the modulation of protein kinase C (PKC) by lead. We examined the effects of inorganic lead on an in vitro model of central nervous system endothelial differentiation in which astroglial cells induce central nervous system endothelial cells to form capillary-like structures. Capillary-like structure formation within C6 astroglial-endothelial cocultures was inhibited by lead acetate with 50% maximal inhibition at 0.5 microM total lead. Inhibition was independent of effects on cell viability or growth. Under conditions that inhibited capillary-like structure formation, we found that lead increased membrane-associated PKC in both C6 astroglial and endothelial cells. Prolonged exposure of C6 cells to 5 microM lead for up to 16 h resulted in a time-dependent increase in membranous PKC as determined by immunoblot analysis. Membranous PKC increased after 5-h exposures to as little as 50 nM lead and was maximal at approximately 1 microM. Phorbol esters were used to determine whether PKC modulation was causally related to the inhibition of endothelial differentiation by lead. Phorbol 12-myristate 13-acetate (10 nM) inhibited capillary-like structure formation by 65 +/- 5%, whereas 4 alpha-phorbol 12,13-didecanoate was without effect. These findings suggest that inorganic lead induces cerebral microvessel dysfunction by interfering with PKC modulation in microvascular endothelial or perivascular astroglial cells.

Animals↗
PubMed1992

Vascular differentiation and glucose transporter expression in rat gliomas: effects of steroids.

The GLUT1 isoform of the glucose transporter is normally expressed at high levels in differentiated brain vessels that also express a permeability barrier. In contrast, malignant brain neoplasms have relatively undifferentiated vessels that are highly permeable, proliferate to high vascular densities, and often lose GLUT1 expression. Using the rat intracerebral 9L glioma model, we investigated whether dexamethasone-induced changes in permeability are associated with the appearance of other differentiated vascular properties. The percentage of vessels expressing immunohistochemically detectable GLUT1 (74.2 +/- 6.1%) and the tumor vessel density as assessed by laminin immunostaining (282 +/- 37 vessels/mm2) did not vary with control tumor size. Dexamethasone treatment resulted in an 83% reduction of vascular permeability to intravenous Evans blue, an increased percentage of vessels expressing GLUT1 (106.4 +/- 10.5%), lower vascular density (102 +/- 64 vessels/mm2), and smaller tumor size (control cross-sectional area, 17.0 +/- 3.4 mm2; treated, 4.6 +/- 1.0 mm2). Essentially all vessels became GLUT1-positive after dexamethasone treatment. Increased GLUT1 expression by glioma vessels in association with the appearance of other signs of differentiation (low vascular density, slow tumor growth) suggests that immunostaining for GLUT1 may identify neoplasms that are biologically less aggressive.

Animals↗
PubMed1992

Effect of elevated ambient glucose upon polyphosphoinositide turnover in bovine retinal endothelial cells and rat astrocytes.

We investigated the turnover of polyphosphoinositides in bovine retinal microvascular endothelial cells and rat astrocytes cultured in the presence of high ambient concentrations of glucose in order to study the possible involvement of this pathway in the pathogenesis of diabetic retinopathy. a 35-45% decrease in the amount of 32P incorporated into phosphatidylinositol(4)phosphate (PIP) and phosphatidyl-inositol(4,5)biphosphate (PIP2) occurred in rat astrocytes but not bovine retinal endothelial cells grown for 14 +/- 3 days in a medium with an elevated (28 mM) glucose concentration. Incorporation of 32P into phosphatidylinositol, phosphatidylcholine, phosphatidylserine and phosphatidylethanolamine was not altered by these conditions. A 39-45% decrease in 32P incorporated into PIP/PIP2 was also found in rat astrocytes grown in 28 mM glucose which were detergent solubilized and incubated with [32P]ATP. Exposure to elevated concentrations of glucose decreased the amount of PIP/PIP2 cleaved by ionomycin or fluoroaluminate treatment, but did not disturb phospholipase C activity. Thus, the lower level of PIP/PIP2, induced by exposure to elevated concentrations of glucose, appears due to changes in phospholipid substrate levels, or polyphosphoinositide kinase activity, rather than a decrease in ATP levels or phospholipase C activity. These results suggest that high ambient glucose levels alter second-messenger generation by astrocytes. In turn, cellular interactions dependent upon these second messengers and important for maintenance of normal microvessel function in the retina may be disrupted.

Animals↗
PubMed1992

Gamma-glutamyl transpeptidase activity in brain microvessels exhibits regional heterogeneity.

Brain microvessels form a tight blood-tissue permeability barrier and express high levels of specific enzymes, including gamma-glutamyl transpeptidase (GGTP). This differentiation is thought to be induced by perivascular astrocytes. By using histochemical methods, we found that the percentage of GGTP-positive vessels varied considerably in different areas of rat brain. Enzyme activity was not found in the pineal gland or the median eminence, where the blood-brain barrier is not expressed. In areas where the blood-brain barrier is expressed, the percentage of GGTP-positive vessels varied from 8% in the optic nerve to 100% in the anterior commissure. The neocortex showed a lower percentage of GGTP-positive vessels (2-15%) than anterior olfactory nucleus (42%), subiculum (70%), hippocampus (48%), and striatum (50-58%). Alkaline phosphatase, another brain microvessel-enriched enzyme, did not show these marked regional differences. The morphometric histochemical results were verified by enzymatic assays in homogenates of different regions from rat and bovine brain and in microvessel preparations of bovine putamen and neocortex. During the postnatal development of rat brain, the difference between neocortex and striatum appeared after day 20. The regional heterogeneity of brain microvessels may be caused by astrocytic heterogeneity and reflect regional heterogeneity in microvascular function.

Aging↗
PubMed1992

Steroid inhibition of neural microvessel morphogenesis in vitro: receptor mediation and astroglial dependence.

Steroid hormones alter several aspects of microvascular function within the CNS. Both microvessel formation and blood-brain barrier expression appear to be influenced by interactions between astrocytes and endothelial cells. To determine if steroids alter astrocyte-endothelial interactions, we studied their effects on astroglial-induced microvessel morphogenesis in vitro. C6 astroglial cells induce bovine retinal microvascular endothelial cells to differentiate into capillary-like structures. Dexamethasone, hydrocortisone, and progesterone at 10 nM inhibited C6-induced microvessel morphogenesis by 75, 35, and 30%, respectively. Inhibition by dexamethasone was both time and concentration dependent, reaching 80-100% at 1 microM. Tetrahydrocortisone and 17 alpha-hydroxyprogesterone had only marginal inhibitory effects. Cortexolone, a glucocorticoid receptor antagonist, blocked inhibition by dexamethasone. Progesterone receptors were expressed in C6 but not bovine retinal microvascular endothelial cells, identifying the astroglial cell as the likely effector of progesterone-mediated inhibition. Astroglial cells were further implicated as the effectors of steroid-mediated inhibition because none of the steroids inhibited astroglial-independent capillary-like structure formation in response to a reconstituted extracellular matrix, Matrigel. These findings are evidence that steroids modulate neural microvascular endothelial cell functions indirectly through perivascular astrocytes via a receptor-mediated mechanism.

Animals↗
PubMed1992

Retinal microvessels express less gamma-glutamyl transpeptidase than brain microvessels.

In this investigation we localized and compared the level of gamma-glutamyl transpeptidase (GGTP) activity in retinal and brain preparations using histochemical, enzymatic and in situ hybridization assays. We compared GGTP distribution to another microvessel specific enzyme, alkaline phosphatase (AP). In the rat brain, GGTP activity was observed in microvessels and choroid plexus by a histochemical method. Similar studies in the rat retina revealed activity in the pigment epithelium but only a very weak reaction in microvessels. Histochemical staining for alkaline phosphatase was observed in both retinal and brain microvessels choroid plexus and pigment epithelium. Biochemical analysis verified that GGTP activity was significantly lower in retinal than brain microvessels, while alkaline phosphatase activity was similar in both types of microvessels. GGTP specific activity of bovine brain and retinal microvessels was 185 +/- 39 mUnits and 8.5 +/- 1.5 mUnits (p less than 0.001), respectively. By contrast, alkaline phosphatase specific activity in brain and retinal microvessels was 732 +/- 139 and 471 +/- 114 (p greater than 0.1), respectively. Choroid plexus and retinal pigment epithelium exhibited similar levels of GGTP and alkaline phosphatase. Differences in GGTP expression between retinal and brain microvessels were also observed on the mRNA level. In situ hybridization studies revealed that brain microvessels expressed four times more GGTP specific mRNA than retinal microvessels. We conclude that retinal microvessels do not express high levels of GGTP which may make them more vulnerable than brain microvessels to injuries mediated by leukotrienes and oxidative stress.

Alkaline Phosphatase↗
PubMed1992

Vascular expression of glucose transporter in experimental brain neoplasms.

Vascular abnormalities in brain neoplasms are important to tumor biology and therapy. Glucose transporter (GLUT1) expression is a differentiated property of normal cerebral microvessels typically associated with expression of the blood-brain barrier. We investigated the relationship of GLUT1 expression to other vascular characteristics in F98, 9L, and C6 gliomas and Walker 256 carcinomas implanted into adult rat brains. The percentages of microvessels with immunohistochemically detectable GLUT1 were 95.5 +/- 3.9 in F98, 60.9 +/- 3.9 in 9L, 45.4 +/- 5.6 in C6, and 1.2 +/- 0.3 in Walker 256 (mean +/- SEM). The percentage of GLUT1-positive vessels in F98 was not statistically different from that in normal brain. GLUT1 expression was not dependent on restricted permeability as all tumors were highly permeable to Evans blue. GLUT1 expression was unrelated to vascular density, vascular morphology, and parenchymal GFAP expression. The expression of GLUT1, a marker of cerebral endothelial differentiation, is a newly described property of glial tumor vessels that may have diagnostic and prognostic significance.

Animals↗
PubMed1991

Mechanisms of lead neurotoxicity.

During the past several years, there has been a renewed interest in the mechanisms by which lead poisoning disrupts brain function. In part, this is related to clinical observations that imply an absence of threshold for toxicity in the immature brain. Many of the neurotoxic effects of lead appear related to the ability of lead to mimic or in some cases inhibit the action of calcium as a regulator of cell function. At a neuronal level, exposure to lead alters the release of neurotransmitter from presynaptic nerve endings. Spontaneous release is enhanced and evoked release is inhibited. The former may be due to activation of protein kinases in the nerve endings and the latter to blockade of voltage-dependent calcium channels. This disruption of neuronal activity may, in turn, alter the developmental processes of synapse formation and result in a less efficient brain with cognitive deficits. Brain homeostatic mechanisms are disrupted by exposure to higher levels of lead. The final pathway appears to be a breakdown in the blood-brain barrier. Again, the ability of lead to mimic or mobilize calcium and activate protein kinases may alter the behavior of endothelial cells in immature brain and disrupt the barrier. In addition to a direct toxic effect upon the endothelial cells, lead may alter indirectly the microvasculature by damaging the astrocytes that provide signals for the maintenance of blood-brain barrier integrity.

Animals↗
PubMed1991

Astroglial-induced in vitro angiogenesis: requirements for RNA and protein synthesis.

Astrocytes are believed to affect microvascular endothelial cell differentiation in brain and retina. Bovine retinal microvessel endothelial cells formed capillary-like structures when cocultured with C6 astroglial cells or in the absence of C6 cells in response to the reconstituted basement membrane protein Matrigel. Using quantitative computer-assisted image analysis, the requirements for RNA and protein synthesis in these two complementary models of in vitro microvessel morphogenesis were examined. Astroglial-dependent capillary-like structure formation was inhibited by up to 87% in a dose-dependent fashion by cycloheximide (0.01-0.1 micrograms/ml), puromycin (0.1-0.25 micrograms/ml), and actinomycin D (0.01-0.025 micrograms/ml). In contrast, the astroglial-independent process in response to Matrigel was not affected by these metabolic inhibitors. These findings suggest that capillary-like structures form in response to astroglial cells in two distinct sequential stages. The first consists of inductive astroglial-endothelial interactions requiring both RNA and protein synthesis. This initiates endogenous endothelial morphogenic events that do not appear to require RNA or protein synthesis, consistent with posttranslational regulatory mechanisms. The first astroglial-dependent step is relevant to the regulation of microvessel formation in brain and retina, whereas the second may represent a morphogenic pathway common to microvessel formation in many tissues.

Animals↗
PubMed1990

Myo-inositol transport into endothelial cells derived from nervous system microvessels.

Myo-inositol, the precursor in the biosynthesis of inositol phospholipids and inositol phosphates, is found in many tissues at concentrations well above its concentration in the plasma, but the highest concentrations are in the central nervous system and the neuroretina. We describe an active, sodium gradient-dependent transport of myo-inositol into cultured endothelial cells derived from bovine retinal microvessels. Transport is inhibited by cytochalasin B, and phloridzin greater than phloretin. Mannitol, sorbitol, and fructose do not inhibit uptake, but D-galactose. inhibits uptake greater than L-glucose greater than D-glucose. The apparent Km of this transport system is 311 +/- 47 (S.D.) microM and the apparent Vmax is 40.8 +/- 2.8 (S.D.) pmol.mg protein-1.min-1. This transport system may be a key in the maintenance of this tissue concentrations as it could concentrate myo-inositol from the plasma into the extracellular spaces of the eye and central nervous system.

Animals↗
PubMed1990

The glucose transporter and blood-brain barrier of human brain tumors.

The glucose transporter of the human brain has been localized to endothelial cells expressing the blood-brain barrier, but little is known regarding its mechanism of induction or whether its expression is exclusively linked with restricted vascular permeability. We investigated glucose transporter expression by vessels in human astrocytic tumors and pulmonary metastases to the brain using immunohistochemical techniques. Vessels in 9 of 10 low-grade astrocytomas and 8 of 10 anaplastic astrocytomas were positive for glucose transporter. Glioblastoma vessels were transporter-positive in only 2 of 10 specimens. Vessels in all three metastatic tumors were negative for the glucose transporter. The decrease in transporter expression observed in higher-grade tumors occurred independently of increases in vascular permeability. In low-grade astrocytomas and glioblastomas transporter expression and contrast enhancement were inversely related, but vessels in 6 of 9 anaplastic astrocytomas were transporter-positive despite contrast enhancement. These findings suggest that separate mechanisms induce the glucose transporter and the permeability restrictions of the human blood-brain barrier. They also have potential implications for the therapy and prognosis of astroglial neoplasms.

Adult↗
PubMed1990

Astrocytes induce neural microvascular endothelial cells to form capillary-like structures in vitro.

Astrocytes maintain a unique association with the central nervous system microvasculature and are thought to play a role in neural microvessel formation and differentiation. We investigated the influence of astroglial cells on neural microvascular endothelial differentiation in vitro. Using an astroglial-endothelial coculture system, rat brain astrocytes and C6 cells of astroglial lineage are shown to induce bovine retinal microvascular endothelial (BRE) cells to form capillary-like structures. Light microscopic evidence for endothelial reorganization began within 48 hours and was complete 72-96 hours following the addition of BRE cells to 1-day-old astroglial cultures. The extent of BRE reorganization was quantitated by computer-assisted analysis and shown to be dependent upon the density of both the BRE and C6 cells within the cocultures. Coculture conditions in which BRE cells were separated from C6 cells by porous membranes failed to generate this endothelial cell change. Likewise, C6-conditioned media and C6-endothelial coculture conditioned media did not induce BRE cell reorganization. Extracellular laminin within the C6-endothelial cocultures, identified by indirect immunofluorescence, was concentrated at the endothelial-astroglial interface of capillary-like structures consistent with incipient basement membrane formation. Astroglial cells accumulated adjacent to capillary-like structures suggesting the presence of bidirectional influences between the reorganized endothelial cells and astroglia. This is the first demonstration of astroglial induction of angiogenesis in vitro and these findings support a functional role for perivascular astrocytes in the vascularization of neural tissue such as retina and brain.

Animals↗
PubMed1990

Phosphoinositide metabolism and prostacyclin formation in retinal microvascular endothelium: stimulation by adenine nucleotides.

Phosphoinositide lipid metabolism and prostacyclin production are implicated in endothelium dependent vascular relaxation in large blood vessels. To determine if these biochemical pathways might be involved in the regulation of microvascular tone in the retina, we measured the formation of 6-keto-prostaglandin-F1 alpha, the stable end product of prostacyclin, and inositol phosphates from 3H-labeled phosphoinositide lipids, in endothelial cells prepared from bovine retinal microvessels and maintained in long-term culture. We found that adenosine 5'-triphosphate and adenosine 5'-diphosphate both stimulated a dose-dependent accumulation of inositol phosphates and of 6-keto-prostaglandin-F1 alpha in these cells. The agonist specificity of the responses, with stimulation by adenosine 5'-triphosphate and adenosine 5'-diphosphate, and inactivity of adenosine 5'-monophosphate and adenosine, suggest that they are mediated through P2 purinergic receptors. The similar early time courses of 6-keto-prostaglandin-F1 alpha and inositol triphosphate production support the hypothesis that prostacyclin formation could result from the mobilization of intracellular calcium by inositol triphosphate, which activates phospholipase A, and thereby releases arachidonic acid to form prostacyclin. These findings point to a role for these cells in the regulation of normal retinal vascular tone. Because phosphoinositide lipid metabolism is altered in diabetes, dysfunction of these biochemical pathways in retinal endothelium could underlie the pathophysiology of diabetic retinopathy.

6-Ketoprostaglandin F1 alpha↗
PubMed1990

Lead poisoning and brain cell function.

Exposure to excessive amounts of inorganic lead during the toddler years may produce lasting adverse effects upon brain function. Maximal ingestion of lead occurs at an age when major changes are occurring in the density of brain synaptic connections. The developmental reorganization of synapses is, in part, mediated by protein kinases, and these enzymes are particularly sensitive to stimulation by lead. By inappropriately activating specific protein kinases, lead poisoning may disrupt the development of neural networks without producing overt pathological alterations. The blood-brain barrier is another potential vulnerable site for the neurotoxic action of lead. Protein kinases appear to regulate the development of brain capillaries and the expression of the blood-brain barrier properties. Stimulation of protein kinase by lead may disrupt barrier development and alter the precise regulation of the neuronal environment that is required for normal brain function. Together, these findings suggest that the sensitivity of protein kinases to lead may in part underlie the brain dysfunction observed in children poisoned by this toxicant.

Blood-Brain Barrier↗
PubMed1990

Intellective functioning and strategy use in children with insulin-dependent diabetes mellitus.

The cognitive development of children with either early or late onset insulin-dependent diabetes mellitus (IDDM) was investigated with tasks measuring intellectual ability, memory, and academic progress. In addition, children's perceptions of their competence and parents' perspectives on family functioning and their children's behavior were compared. It was found that children with IDDM scored within the normal range on standardized measures of intelligence and academic performance but evidenced some school difficulties, as reflected in subscale performance as well as in their need of remedial education services. Further, evidence was found to suggest deficiencies in children's use of strategies to organize and recall information, particularly for those with early onset of disease. Children's perceived self-competencies and parents' reports of family functioning were strikingly similar across groups. However, parents of those children whose illness began prior to age 5 reported their children to have poor attention spans and difficulty completing tasks.

Achievement↗
PubMed1989

Use of intravenous immune globulin in the treatment of seizure disorders.

Recurrent seizures are a significant cause of childhood morbidity. Often the underlying pathogenesis is unknown. Studies from several laboratories have suggested a relationship between perturbations of humoral immunity and clinical epilepsy. Brain-specific antigens have been observed in the systemic circulation after brain injury. This can be followed by the appearance of antibodies to neuronal tissues in both serum and cerebrospinal fluid. Moreover, there appears to be disproportionate representation of persons with coexisting humoral immune deficiencies among patients with chronic epilepsy. Interestingly, a potent immunosuppressant, adrenocorticotropic hormone, has long been the principal treatment for infantile spasms. Given this body of information associating humoral immunity with recurrent seizures, several investigators have recently attempted to treat intractable childhood epilepsy with high-dose intravenous immune globulin. Although the number of patients treated is small and it is not possible to make comparisons between individual studies, nevertheless the results have been encouraging. Five patients with intractable seizures who failed to respond to conventional therapy were treated with an outpatient regimen of intravenous immune globulin (1 gm/kg/day) for 2 days. Two patients showed significant clinical improvement while a third had an equivocal response. Immunologic analysis of these patients before therapy revealed a surprisingly high incidence of immune abnormalities. We conclude that intravenous immune globulin may be useful in the treatment of selected patients with chronic epilepsy unresponsive to anticonvulsants.

Adolescent↗