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E L White

Publications and source records attributed to E L White.

At least 73 records · Page 4Linked to original sources

Quantification of thalamocortical synapses with spiny stellate neurons in layer IV of mouse somatosensory cortex.

The distribution of thalamocortical (TC) and other synapses involving spiny stellate neurons in layer IV of the barrel region of mouse primary somatosensory cortex (SmI) was examined in seven male CD/1 mice. TC axon terminals were labeled by lesion-induced degeneration, which has been shown to label reliably all TC synapses in mouse barrel cortex. Spiny stellate neurons, labeled by Golgi impregnation and gold toning, were identified with the light microscope prior to thin sectioning and electron microscopy. Analysis of eight dendritic segments from seven spiny stellate neurons showed that most of their synapses are with their dendritic spines, rather than with their shafts. Axospinous synapses are primarily of the asymmetrical type, whereas axodendritic synapses are mainly of the symmetrical type. Dendrites of spiny stellate neurons consistently form thalamocortical synapses, most of which involve spine heads rather than spine stalks or dendritic shafts. From 10.4% to 22.9% of all asymmetrical synapses with dendrites of spiny stellate neurons involve TC axon terminals. In general, this is a higher range than the ranges that characterize the TC synaptic connectivity of dendrites belonging to other types of neurons, implying that spiny stellate neurons are perhaps more strongly influenced by TC synaptic input than other types of cortical neurons examined previously. Spines involved in TC synapses were distributed irregularly along each of the stellate cell dendrites; about half of the interspinous intervals between these spines were about 5 microns or less. Modulations of the efficacy of TC synaptic input to dendrites of layer IV spiny stellate neurons are discussed in the light of recently reported computer simulated analyses of axospinous synaptic connections.

Animals↗

Synaptic connections of callosal projection neurons in the vibrissal region of mouse primary motor cortex: an electron microscopic/horseradish peroxidase study.

Reciprocal axonal projections between homotypic areas of the vibrissal region of mouse primary motor cortex (MsI) (Porter and White: Neurosci. Lett. 47:37-40, '84) suggested the existence of reciprocal synaptic connections between callosal projection neurons and callosal afferents. In the present study, the retrograde transport of horseradish peroxidase (HRP) was combined with lesion-induced degeneration to identify synapses between callosal afferents and callosal neurons in the corresponding region of the contralateral cortex. The procedure was as follows: MsI was injected with HRP and aspirated on the following day. After 4 days, the animals were perfused and motor cortex was processed for HRP according to a variation of the Adams (Brain Res. 176:33-47,'77) technique, and postfixed in OsO4. The methods used consistently filled fine dendritic branches and spines with dense reaction product, thus allowing examination of synaptic contacts with these processes. All callosal projection neurons were identified as pyramidal neurons, having somata in cortical layers II/III and V. Labeled cells from each of the two levels were prepared for electron microscopy, and that part of each cell's apical dendrite that traversed the superficial cortical layers, where most callosal axons terminate, was cut in an unbroken series of thin sections. Micrographs were taken of all labeled profiles in each thin section, and tracings of the profiles were assembled to reconstruct the apical dendrites. Data on the distribution, type, and amount of callosal and other synapses with the shaft and spines of the apical dendrites were obtained by examining the reconstructions. In addition, profiles of basal dendrites of layer II/III cells were examined in thin sections to ascertain the numbers of callosal and other synapses formed with their shafts and spines. The proportion of synapses that each dendrite formed with callosal axon terminals was compared to the concentration of callosal afferents in the neuropil. Dendrites of both layer II/III and layer V pyramidal cells synapsed with callosal axon terminals. The apical and basal dendrites of layer II/III neurons formed a similar proportion of their synapses with callosal afferents, and this was similar to the concentration of callosal synapses in the surrounding neuropil. Segments of apical dendrites belonging to layer V and layer II/III neurons course through neuropil containing nearly the same concentration of degenerating callosal terminals, but the layer V cells form fewer callosal synapses.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Distribution of glutamic acid decarboxylase-immunoreactive structures in the barrel region of mouse somatosensory cortex.

Neuronal structures in the barrel region of the mouse primary somatosensory cortex containing gamma-aminobutyric acid (GABA) were identified by an immunocytochemical method, using an antiserum to glutamic acid decarboxylase (GAD), the rate-limiting enzyme in the synthesis of GABA. GAD-positive cell bodies were found in all layers of the barrel cortex, but were more concentrated in the upper portion of layer II/III, and in layers IV and VI. Puncta, presumably axon terminals, were also distributed throughout the cortical layers; a high density of puncta occurs in layer IV, whereas a somewhat lower density characterizes layer VI. Based on the shapes of their somata and the distribution of their dendrites it was determined that all GAD-positive cell bodies were of the non-pyramidal type.

Animals↗

Specific methods to identify plasma binding abnormalities in euthyroid hyperthyroxinemia.

Methods to identify the plasma T4-binding abnormalities that can cause euthyroid hyperthyroxinemia were evaluated in patients with excess T4-binding globulin, familial dysalbuminemic hyperthyroxinemia, prealbumin-associated hyperthyroxinemia, and autoantibody binding of T4. Familial dysalbuminemic hyperthyroxinemic serum showed a unique persistence of abnormal [125I]T4 binding when diluted 1:100 in phosphate buffer with added 1000-fold excess of unlabeled T4 (10(-6) M T4). Immunoprecipitation of [125I]T4 by antibody to prealbumin, precipitation of [125I]T4 by polyethylene glycol 6000 19%, and in vitro resin uptake of T3 were specific for prealbumin-associated hyperthyroxinemia, autoantibody binding of T4, and T4-binding globulin excess, respectively. These simple methods facilitate investigation of patients with euthyroid hyperthyroxinemia and will identify individuals and families at risk of misdiagnosis by standard methods. Use of these techniques rules out the known binding abnormalities in hyperthyroxinemic patients and may make the diagnosis of generalized hormone resistance more specific.

Humans↗

The identification of thalamocortical axon terminals in barrels of mouse Sml cortex using immunohistochemistry of anterogradely transported lectin (Phaseolus vulgaris-leucoagglutinin).

The anterograde transport and immunohistochemical demonstration of the lectin, Phaseolus vulgaris-leucoagglutinin (PHA-L) has been used to label thalamocortical axon terminals in barrels of mouse SmI cortex. The reaction product is visible with both the light and electron microscopes so that the distribution of axons and the types of synapses they form can be determined.

Afferent Pathways↗

The use of lectin transport in the mouse central nervous system as an anterograde axonal marker for electron microscopy.

Lesion-induced axonal degeneration and autoradiography-electron microscopy have been the only reliable anterograde axonal markers available for electron microscopic examination of neuronal circuitry. However, these methods have their limitations. Recently, Phaseolus vulgaris-leucoagglutinin (PHA-L) has been used as an anterograde axonal marker for light microscopy. This report describes the use of this lectin as an anterograde marker for electron microscopy. PHA-L was injected into mouse SmI cortex or ventrobasal thalamus. Using standard immunohistochemical techniques, the transported lectin was tagged with antibody, which was then visualized with avidin-biotin-horseradish peroxidase binding. Light microscopy demonstrated anterograde transport to predicted cortical regions. With the electron microscope, labeled axon terminals were seen forming asymmetric synapses with spines, dendrites and cell bodies.

Afferent Pathways↗

Thalamocortical and other synapses involving nonspiny multipolar cells of mouse SmI cortex.

Golgi-impregnated and -deimpregnated neurons having somata in layer IV of mouse posteromedial barrel subfield (PMBSF) cortex were identified with the light microscope and then extensive portions of them were examined with the electron microscope. Dendrites of nine nonspiny multipolar cells and eight of their cell bodies were reconstructed from serial thin sections to determine the numbers and types of symmetrical, asymmetrical, and thalamocortical synapses they formed. Results of this analysis show that cells of the same general morphological class may form widely different patterns of synaptic connections: some nonspiny multipolar cells had dendrites that formed a high proportion of their synapses with thalamocortical axon terminals, whereas dendrites belonging to other cells formed only very small proportions of thalamocortical synapses. A similar diversity characterized the synaptic connections of cell bodies: some formed more symmetrical than asymmetrical synapses, others the reverse. Some formed high proportions of thalamocortical synapses, others much less. Comparisons of thalamocortical synaptic input to cell bodies and dendrites showed that one cell formed about the same proportions of thalamocortical synapses with its cell body as with its dendrites. For two other cells the proportions of thalamocortical synapses formed with their somata was about double that formed with their dendrites. The remaining five cell bodies examined formed far higher proportions of thalamocortical synapses than did their dendrites. That different nonspiny multipolar cells form such contrasting synaptic patterns suggests that included within this morphological classification are cells which are likely to have very different functional roles.

Animals↗

Termination of callosal afferents onto identified callosal projection neurons in the primary motor cortex of the mouse.

Techniques utilizing the retrograde transport of horseradish peroxidase and lesion-induced degeneration have been used to identify neurons which form the callosal projection pathway and callosal axon terminals in the primary motor cortex of the mouse. Synapses occur between callosal projection neurons and degenerating callosal afferents, demonstrating a direct synaptic link between neurons which project to homotopic regions of the contralateral cortex.

Animals↗

A pituitary tumor producing high molecular weight adrenocorticotropin-related peptides: clinical and cell culture studies.

The tissue-specific processing of proopiomelanocortin (POMC), the precursor of ACTH, beta-endorphin, and their related peptides, is currently of considerable interest. We report a patient with a large aggressive pituitary tumor, Cushing's syndrome, and hyperpigmentation managed by transsphenoidal hypophysectomy, bilateral adrenalectomy, and sellar irradiation. Preoperatively, plasma levels of immunoreactive ACTH (ir-ACTH; 280 ng/liter) and beta-endorphin (ir-beta EP; 520 ng/liter) were moderately elevated. Chromatography of the plasma showed two peaks of ACTH immunoreactivity, with the major peak eluting in the void volume, and two major peaks of ir-beta EP, corresponding to the elution positions of beta-lipotropin and beta-endorphin standards. Plasma ir-ACTH and ir-beta EP were not suppressed by high doses of glucocorticoid or bromocriptine, a degree of autonomy more commonly found with POMC production from ectopic sources than that from pituitary tumors. Tissue removed at operation was enzymatically dispersed, and the cells were cultured in suspension, propagated, and passaged sequentially for over 20 passages. Using this cell line, we demonstrated that the biosynthesis of POMC, its pattern of processing, and the release of POMC/ir-beta EP/ir-ACTH in vitro were consistent with the in vivo evidence of autonomous secretion and abnormal processing of POMC by this pituitary tumor.

Adenoma↗

Evaluation of free thyroxine methods in the presence of iodothyronine-binding autoantibodies.

Circulating iodothyronine-binding autoantibodies interfere with total T4 and T3 RIAs, giving falsely high or falsely low values depending on the assay separation method used. Direct serum free T4 (FT4) measurement should compensate for such abnormal binding if the method is free of artefact. We assessed 5 different FT4 methods in 12 patients with immunoglobulin binding of iodothyronines, with limited assessment of a sixth method in 2 subjects. Thyroid status was assessed clinically and by measurement of TSH and its response to TRH. T4 methods which use analog tracers, i.e. Amerlex, and Clinical Assays One-step FT4, gave spuriously high values in almost all hypothyroid and some euthyroid patients, due to immunoglobulin binding of the tracers used in these techniques. The kinetic indirect method (Corning Immunophase FT4) gave inappropriately high values in 4 of 6 hypothyroid patients. FT4 by equilibrium dialysis or by adsorption chromatography and RIA, accurately assessed thyroid status. These findings suggest that FT4 methods are valid in patients with circulating iodothyronine-binding immunoglobulins only if the free hormone fraction is physically separated from serum binding proteins before the assay procedure.

Adult↗

An evaluation of a water purification system for use in animal facilities.

A commercially available water purification system was evaluated for its ability to minimize chemical and microbial contaminants. The reduction or removal of these impurities from the drinking water of experimental animals would reduce experimental variability. 3 strains of bacteria were collected from the processed water. An increase in the total number of bacteria was observed the longer the filters remained in use. Determinations of heavy metals in water samples before and after processing were made for lead, zinc, copper, nickel, manganese, iron, arsenic and mercury. Calcium and magnesium levels were also determined. The concentrations of these inorganic chemicals were reduced by the purification process except at 2 time points in which desorption of the chemical could have occurred. Bacterial colonization and desorption of these chemicals were controlled by installing new filter cartridges. Volatile halocarbon concentrations were determined for water samples before and after purification. All volatile halocarbons analyzed were less than 10 ppb before and after purification at all time points. Other organic chemicals were greatly reduced by the purification process. In a study of contaminants associated with installation of the unit, it was found that flushing the unit for 8 days reduced lead and methyl ethyl ketone concentrations to insignificant levels. The purification system was found to be effective in providing high quality drinking water as verified by a microbial and chemical testing program.

Animals↗

Afferent and efferent pathways of the vibrissal region of primary motor cortex in the mouse.

The afferent and efferent connections of the vibrissal representation within the mouse primary motor cortex (MsI) were identified by using the retrograde transport of horseradish peroxidase (HRP) and the anterograde transport of tritiated amino acids injected into MsI. Following aldehyde perfusion brains were frozen-sectioned at 40 microns and reacted for HRP using the 3-3' diaminobenzidine-cobalt chloride technique of Adams ('77). Alternate HRP reacted sections were processed for autoradiography. HRP-filled pyramidal cell somata and concentrations of developed silver grains above background levels were observed in both the vibrissal area of primary somatosensory cortex (SmI) cortex (i.e., the posteromedial barrel subfield; PMBSF cortex) and in the face region of SmII (area 40). In both regions labeled somata occurred predominantly in cortical layers II-III and V. Autoradiographic label was superimposed over the regions containing labeled somata but exhibited a less distinct laminar organization. A dense reciprocal projection connected the injection site with the homotopic area in contralateral MsI; somata occurred for the most part in layers III and V. Developed silver grains were uniformly dispersed over the area containing labeled cell bodies. HRP-labeled pyramidal somata were noted in contralateral PMBSF cortex, but no silver grains occurred in this region. Reciprocal projections linked MsI cortex with the ipsilateral thalamic nuclei: ventralis pars lateralis (VL) and centralis pars lateralis (CL) and with the zona incerta (ZI). Labeled cell bodies and developed silver grains were more dense in VL than in CL. The ipsilateral striatum and thalamic reticular nucleus (NRT) received afferents from the motor cortex but did not project to it. Thus, the vibrissal area of primary motor cortex is connected with a number of cortical and subcortical structures, each of which has been shown to play a role in motor performance. Identification of the afferent and efferent pathways of MsI cortex will now enable further investigation of the ultrastructural and synaptic organization of the vibrissal area of MsI.

Afferent Pathways↗

"Unbound analog" radioimmunoassays for free thyroxin measure the albumin-bound hormone fraction.

We have assessed the influence of albumin-bound thyroxin (T4) on apparent free T4 values obtained by two "unbound analog" free T4 methods (AmerlexR Free T4 and Clinical Assays one-step Free T4). We evaluated sera showing three different albumin anomalies: total hereditary analbuminemia, partially corrected analbuminemia, and familial dysalbuminemic hyperthyroxinemia, where abnormal albumin-binding of analog tracer is associated with high apparent free T4 values by these methods. In hereditary analbuminemia, free T4 was almost undetectable by both assays; in contrast, free T4 by equilibrium dialysis was normal. After addition of T4-free human serum albumin, the apparent free T4 concentration in total hereditary analbuminemia became normal by the analog methods. Immunoprecipitation of [125I]T4 and the unidentified labeled kit analogs by antiserum to human albumin was negligible in untreated total hereditary analbuminemia and approximately twice normal in familial dysalbuminemic hyperthyroxinemia. Therefore, alterations in tracer binding to albumin correlate with the apparent free T4 concentrations obtained by the analog methods. The interactions of the unidentified analog tracers and T4 with albumin are such that these techniques principally reflect the albumin-bound T4 moiety.

Antigen-Antibody Complex↗

A quantitative study of the thalamocortical and other synapses in layer IV of pyramidal cells projecting from mouse SmI cortex to the caudate-putamen nucleus.

The thalamocortical and other synapses of the apical dendrites of corticostriatal projection neurons in mouse primary somatosensory cortex (SmI) were examined by combining anterograde degeneration with the retrograde transport of horseradish peroxidase (HRP). Electrolytic lesions were made in the ventrobasal thalamus, followed 3 days later by injections of 40% HRP into the ipsilateral caudate-putamen nucleus. The next day, the mice were perfused and the SmI cortex ipsilateral to the lesion and injection sites was chopped at 125-micrometer and reacted for HRP using a CoCl2-DAB method. HRP-labeled corticostriatal cells in SmI cortex were medium-sized pyramidal cells, having somata located in the superficial portion of layer V and apical dendrites extending into layer I. Seven corticostriatal cells were serially thin sectioned and the layer IV portions of their apical dendrites were reconstructed. Each apical dendrite formed only one or two thalamocortical synapses (0.3 to 0.9% of their synapses in layer IV) indicating that corticostriatal neurons may be minimally responsive to direct synaptic input from the specific thalamic nuclei. Each apical dendrite formed about 12.6 asymmetrical synapses for every symmetrical synapse, suggesting that the relative numbers of excitatory and inhibitory synapses impinging on apical dendrites belonging to an individual class of neurons may be specified.

Afferent Pathways↗

Determination of formaldehyde in biological tissues by gas chromatography/mass spectrometry.

A quantitative method is described for the determination of formaldehyde in biological tissues by stable isotope dilution using gas chromatography/mass spectrometry. (13C2H2)Formaldehyde is used as the isotopic diluent. After tissue homogenization, derivatization is carried out in situ with pentafluorophenylhydrazine, followed by extraction and analysis using selected ion monitoring. The sensitivity of the technique is higher than that of conventional methods of formaldehyde analysis, enabling endogenous formaldehyde to be quantitatively analyzed in tissues, even in samples as small as 20 mg wet weight. The effects of exposure to airborne formaldehyde or to airborne methyl chloride on the formaldehyde concentrations of several tissues of Fischer-344 rats are reported.

Animals↗

A quantitative study of thalamocortical and other synapses involving the apical dendrites of corticothalamic projection cells in mouse SmI cortex.

Lesion-induced degeneration was combined with the retrograde transport of horseradish peroxidase (HRP) to examine the thalamocortical and other synaptical relationships of corticothalamic projection cells in mouse SmI cortex. Injections containing 40% HRP were placed in the ventrobasal thalamus and the next day, electrolytic lesions were made of the injection site. About four days later, the animals were perfused with aldehydes and SmI cortex ipsilateral to the injection and lesion sites was tissue chopped and reacted for HRP. The somata of HRP-filled corticothalamic cells in SmI cortex had diameters of about 10 micrometers and occurred in the upper half of layer VI and in the lower half of layer V; their apical dendrites usually terminated within or just below layer IV, but in some instances, extended nearly to the pial surface. Reconstructions of serial thin sections through the layer IV portions of the apical dendrites of seven well-labelled corticothalamic cells showed them to form 7 to 20% of their synapses with degenerating thalamocortical axon terminals. In contrast, the proportion of thalamocortical synapses formed by the layer IV dendrites of corticostriatal (Hersch & White, in preparation) and corticocortical (White & Hersch, 1981) projection cells ranges from 0.3 to 0.9% and from 1.5 to 6.8%, respectively. We interpret these findings to indicate that pyramidal cell dendrites are specified to form characteristic proportions of thalamocortical synapses.

Animals↗

Familial euthyroid thyroxine excess: characterization of abnormal intermediate affinity thyroxine binding to albumin.

The abnormal high capacity T4 binding site of familial euthyroid T4 excess was separable from prealbumin and T4-binding globulin but not from albumin. We therefore compared T4 binding by albumin preparations isolated from the sera of normal and affected subjects. By equilibrium dialysis, albumin from affected subjects showed an extra T4 binding site (Kd approximately 50 nM) in addition to the T4 binding sites of normal albumin (Kd approximately 4 microM). Comparison of the estimated capacity of the additional site (200 microM) with the molar concentration of albumin suggested that only about one third of albumin molecules from affected subjects contained the extra binding site. Estimates of affinity and capacity were used to derive combining powers for the diverse classes of serum T4 binding sites. From these estimates, it appears that the presence of the abnormal site accounts for the approximate doubling of normal mean total T4 (from approximately 100 nM or 7.7 micrograms/dl to approximately 200 nM or 15.5 micrograms/dl), in order to maintain a normal free T4 in the face of the increased T4 association with albumin. Studies of [125I]T4 displacement from albumin of affected subjects showed low T3 affinity and competition by barbitone. Relative molar concentrations to give equivalent displacement of [125I]T4 were: 3,3',5,5'-tetraiodothyroacetic acid, 0.4; T4, 1.0; rT3, 4; 8-anilinonaphthalene sulfonic acid, 10; T3, 80; salicylate, 200; and barbitone, 40,000. Studies with dithiothreitol suggested that disulfide bonds were critical in maintaining the T4-albumin association. These findings indicate that familial T4 excess is due to abnormal intermediate affinity, sulfhydryl-sensitive T4 binding sites that are inseparable from the albumin found in affected subjects.

Barbital↗