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A E Leure-duPree

Publications and source records attributed to A E Leure-duPree.

13 recordsLinked to original sources

Effects of zinc deficiency on protein synthesis and expression of specific mRNAs in rat liver.

The effects of zinc deficiency on protein synthesis and expression of specific mRNAs were assessed in rat liver. Zinc deficiency had no apparent effect on liver weight, protein content, or RNA content when these properties were compared with values obtained using pair-fed rats. However, zinc deficiency resulted in a lower rate of hepatic protein synthesis. The decreased rate of protein synthesis was due to a decrease in the rate of synthesis of proteins retained in the liver, with no apparent change in the synthesis of secreted proteins. Analysis of expression of specific gene products, as assessed by in vitro translation of total RNA followed by two-dimensional gel analysis, showed that the expression of only a few mRNAs was altered by zinc deficiency. The patterns of change in gene expression resulting from zinc deficiency varied from almost complete repression to full expression. In additional studies, cDNA clones to serum retinol-binding protein and transthyretin were used to examine the effect of zinc deficiency on the relative abundance of mRNA for these two proteins. The relative abundance of mRNA for transthyretin was specifically elevated as a result of zinc deficiency. In contrast, the relative abundance of mRNA for hepatic serum retinol-binding protein was increased in both zinc-deficient and pair-fed rats. Therefore, the observed change in mRNA for serum retinol-binding protein was apparently at least in part due to the inanition that accompanies zinc deficiency. Overall, the results suggest that zinc can regulate the synthesis of specific proteins in rat liver through changes in the relative abundance of specific mRNAs.

Animals↗

Genetic analysis of the major homology region of the Rous sarcoma virus Gag protein.

The mature cores of all retroviruses contain a major structural protein known as the CA (capsid) protein. Although it appears to form a shell around the ribonucleoprotein complex that contains the viral RNA, its function in viral replication is largely unknown. Little sequence similarity exists between the CA proteins of different retroviruses, except for a region of about 20 amino acids termed the major homology region (MHR). To examine the role of the CA protein in particle assembly and release, mutants of Rous sarcoma virus were created in which segments of CA were deleted or single conserved residues in the MHR were altered. The ability of the deletion mutants to release particles at rates similar to the wild-type protein demonstrated that the CA domain of Gag is not an essential component of the minimal budding machinery. Certain point mutations in the MHR region did block assembly and release in certain cell types, presumably by perturbing the global structure of the Gag precursor. Another group of MHR substitutions produced noninfectious or poorly infectious particles that were normal in their content of gag and pol gene products and viral RNA. The mutants were capable of initiating reverse transcription in vitro; however, the association of CA protein with the core was compromised, as indicated by its sensitivity to extraction with nonionic detergent. Prominent blebs on the virion envelope also indicated a disturbance at the membrane. Finally, an anti-peptide serum directed against MHR was found to react with the uncleaved Gag protein but not with mature CA, suggesting that MHR undergoes a dynamic rearrangement upon liberation from the polyprotein. We conclude that the MHR is involved in the very late steps in maturation of the virion (i.e., ones that occur after budding is initiated) and is essential for proper function of the core upon entry into a new host cell.

Amino Acid Sequence↗

Necessity of the spacer peptide between CA and NC in the Rous sarcoma virus gag protein.

A mutant of Rous sarcoma virus was constructed in which the nine amino acids that separate the CA and NC sequences in the Gag protein were deleted. The spacer peptide deletion mutant produced particles containing the normal complement of viral RNA and all of the viral proteins, including reverse transcriptase. Though electron microscopy revealed particles of normal morphology, the particles were noninfectious. The normally slow maturation of the CA protein, which involves cleavage of the spacer peptide from the carboxy terminus, was bypassed in this mutant, and the association between CA and the internal components of the core appears to have been disrupted. The results suggest that the spacer peptide has an essential role in directing folding and/or oligomerization of the CA subunits within the capsid structure.

Amino Acid Sequence↗

Zinc deficiency affects the composition of the rat adrenal gland.

The response of the adrenal gland to zinc deficiency was examined in male weanling rats. In comparison with decapsulated adrenals from ad libitum fed controls, glands from zinc deficient rats had greater relative weight (mg/g body wt), DNA concentration, and total lipid and cholesterol concentrations as well as a smaller protein/DNA ratio. Several of these differences (protein/DNA and cholesterol concentration) could be attributed to the inanition accompanying zinc deficiency, as zinc deficient values were similar to those of pair fed controls. Values for total DNA and protein concentration were similar for all groups. Electron micrographs of the zona fasciculata showed a small number of lipid droplets in the adrenals from ad libitum fed controls, an increase in lipid droplets from pair fed controls, and an even more striking increase in lipid droplets from the zinc deficient adrenals. The increased adrenal lipid composition in the zinc deficient group may be secondary to enhanced steroidogenesis or a zinc deficiency-induced defect of lipid metabolism.

Adrenal Glands↗

The effect of severe zinc deficiency on the morphology of the rat retinal pigment epithelium.

Male Sprague-Dawley weanling rats maintained on a controlled dietary zinc intake had an accumulation of osmiophilic inclusion bodies in the retinal pigment epithelium (RPE) of the zinc-deficient group. At 7 weeks of zinc deficiency there were marked ultrastructural alterations of the RPE. In some instances a deepening of the basal infoldings of the cells of the RPE were observed. There was also vesiculation and degeneration of the photoreceptor outer segments. No changes were observed in the pair-fed, weight-restricted, and ad libitum-fed controls. The possible functions of zinc in the retina are discussed.

Animals↗

Electron-opaque inclusions in the rat retinal pigment epithelium after treatment with chelators of zinc.

Sprague-Dawley male rats given intraperitoneal injections of zinc chelators, namely dithizone and 1,10-phenanthroline, had unusual osmiophilic inclusion bodies in their retinal pigment epithelium (RPE). These structures were not seen in untreated or vehicle-treated controls. The morphology of the inclusion bodies clearly distinguished them from cytoplasmic organelles observed normally in the RPE. The number of inclusion bodies is dose-dependent. The bodies do not have the morphological characteristics of lipid droplets seen infrequently in the untreated rat RPE. The results suggest that sequestration of zinc, either rapidly with injections of dithizone or more slowly with daily administration of 1,10-phenanthroline, exerts pronounced effects on the retinal pigment epithelial cells.

Animals↗

The morphology of extrachoroidal ependyma overlying gray and white matter in the rabbit lateral ventricle.

A morphologic investigation of ependyma over gray matter (caudate nucleus) and over periventricular white matter (tapetum) of the rabbit lateral ventricle was undertaken prior to evaluation of morphological changes which occur with experimental hydrocephalus. Ependymal cells over the caudate nucleus are cuboidal and heavily ciliated. Numerous microvilli cover the cell surface. The lateral margins are straight and interdigitations between adjacent ependymal cells are absent. Ependymal cells over white matter are squamous. Nonciliated as well as ciliated cells contribute to the epithelial lining. Microvilli are present at the cell surface but tend to aggregate near the cellular borders. The lateral margins are convoluted and complex interdigitations are present between adjacent cells. Morphologic differences between ependymal cells over the caudate nucleus and those over periventricular white matter may help to explain the differential response to hydrocephalus observed in these two regions of the lateral ventricle.

Animals↗

Photoreceptors and oil droplet colors in the red area of the pigeon retina.

Six types of photoreceptors in the red area (dorso-temporal quadrant) of the pigeon retina are identified using Golgi impregnation, light microscopy and electron microscopy. Golgi impregnation is used to categorize the receptors into morphological types. Examination of oil droplets in the inner segments of cones in fresh, unfixed tissue shows five different types which can be characterized by color, size and stratification. Therefore, in sections through the length of the receptors examined by electron microscopy, the oil droplets contained in the inner segments of the cones can be identified as to their color by their characteristics (i.e., size and stratification), and the groups of receptors thus classified, further characterized as to the morphology of their terminals. Rods have no oil droplets in their inner segments, and their synaptic terminals are located in the outermost stratum of the outer plexiform layer OPL). Principal members of double cones have yellow oil droplets in their inner segments, while accessory members contain small colorless oil droplets. The synaptic terminals of double cones are located in the same (outermost) stratum of the OPL as rod synaptic terminals. Two types of single, straight cones house either red or orange oil droplets and terminate in the intermediate stratum of the OPL. Oblique single cones with yellow-green oil droplets in their inner segments contribute synaptic terminals to the innermost stratum of the OPL.

Animals↗

The neurohypophyseal capillary bed. II. Specializations within median eminence.

Vascular casts of the pituitary gland, median eminence and hypothalamus from several mammalian species were examined by scanning electron microscopy. These observations were correlated with light microscopic studies of injected, cleared median eminence-pituitary specimens and with light microscopic examination of serial sections of injected hypothalamic, median eminence, and pituitary specimens employing reflected lighting or epi-illumination. Transmission electron microscopy was employed to study long portal vessels on the ventral surface of the rat median eminence. In each of the species examined, the median eminence (infundibular) capillary bed is subdivided into an external and an internal plexus. The external plexus (the neurohemal contact zone) receives an arterial supply from the superior hypophyseal arteries and is continuous with the capillary bed of the infundibular stem and process. Egress from the external plexus is possible via three vascular routes: (a) by fenestrated portal vessels and capillaries to the adenohypophysis, (b) by capillary connections to the medial basilar hypothalamus and, (c) by internal plexus capillaries to the ependyma of the median eminence. Median eminence vasculature is structurally organized to deliver (1) hypothalamic and neurohypophyseal peptides to the glandular pituitary via portal vessels, (2) hypothalamic and pituitary secretions to the medial basilar hypothalamus via capillaries, and (3) hypothalamic and pituitary secretions to distant brain sites through cerebrospinal fluid via ventricular and subarachnoid routes.

Animals↗

Pathophysiology and electron microscopy of melanomalytic glaucoma.

We examined an eye with melanomalytic glaucoma by quantitative aqueous perfusion, light and electron microscopy. Facility of outflow was markedly diminished from normal in vivo and in vitro. The trabecular meshwork was heavily laden on its surface and in the intertrabecular spaces with macrophages. Many of these cells appeared to be wandering macrophages, while some appeared to be detached, phagocytic trabecular endothelial cells.

Aqueous Humor↗

Synaptic organization of the external cuneate nucleus in the rat.

The normal synaptic organization of the rat external cuneate nucleus (ECN) has been investigated. The characteristic feature of the ECN neuropil is complex synaptic arrangements termed synaptic glomeruli. These involve both axo-dendritic and axo-axonic synapses and usually are isolated from the surrounding neuropil by lamellar glial sheaths. Glial isolation of synaptic glomeruli is conspicuous near capillaries. Central glomerular bouton profiles vary in size (2-9 micron diam) and configuration. They form asymmetrical synapses onto dendritic shafts and spines and occasionally receive symmetrical synapses from en passant bouton profiles. Other forms of axo-dendritic synapses not associated with synaptic glomeruli are observed. The type frequently observed is formed by one or more boutons (0.5 4.0 micron diam) synapsing onto a dendrite or dendritic spine. "Isolated" axo-dendritic synapses are surrounded by glial lamellae. Elongated "giant" boutons make multiple synaptic contacts along a dendritic shaft. Serial axo-axonic synapses are found mainly in caudal regions of the nucleus. Axo-somatic synapses are formed by small boutons (0.3.2.0 micron diam) contacting medium (16-24 micron) and small (9-14 micron) neurons. The least frequently observed bouton type in the rat ECN contains numerous granular vesicles, 80-100 nm in diameter. These boutons may contact dendrites, neuronal somas or other boutons.

Animals↗