Biomedical subjects

W Singer

Publications and source records attributed to W Singer.

At least 163 records · Page 9Linked to original sources

PubMed1990

Human pituitary corticotroph adenomas in vitro: morphologic and functional responses to corticotropin-releasing hormone and cortisol.

We examined the direct effects of corticotropin-releasing hormone (CRH) and cortisol on the morphology of cells from 6 functioning human pituitary corticotroph adenomas in culture using both light and electron microscopic morphometry and correlated the structural changes with alterations in adrenocorticotropin (ACTH) release in each case. During incubations lasting 2 or 24 h, ACTH release was increased by CRH and reduced by cortisol. After incubations lasting from 2 to 72 h, light microscopic morphometric analysis showed no significant differences in cell size, nuclear area, cytoplasmic area or nuclear/cytoplasmic ratio between treated and control adenoma cells. Ultrastructural morphometry documented increased cytoplasmic volume density (CVD) of rough endoplasmic reticulum and/or Golgi apparatus and reduced CVD of secretory granules in cells incubated with CRH. There was no consistent change in CVD of endoplasmic reticulum, Golgi apparatus or secretory granules in adenoma cells incubated with cortisol, but in all tumors there were marked filament accumulations indicating a direct effect of cortisol on adenomatous corticotrophs. The changes were similar after 2- and 72-hour exposures. These results indicate that (1) some adenomatous corticotrophs can respond to CRH and cortisol; (2) the morphologic changes observed in cells treated with CRH correlate with increased ACTH release, and (3) accumulation of filaments is the direct effect of cortisol and is associated with reduced ACTH release.

Adenoma↗

PubMed1990

The formation of cooperative cell assemblies in the visual cortex.

During a critical period of postnatal development of the mammalian visual cortex, synaptic connections are susceptible to use-dependent modifications. Synaptic connections strengthen if pre- and postsynaptic elements are active simultaneously and postsynaptic depolarization is sufficient to allow for the activation of N-methyl-D-aspartate (NMDA)-receptor-gated conductances. By contrast, synaptic gain decreases if postsynaptic activation exceeds a critical threshold and presynaptic afferents are not capable of activating NMDA-receptor-dependent conductances. These processes lead to selective stabilization of connections between neuronal elements which often exhibit correlated activity and thus modify connectivity according to functional criteria. It is suggested that such experience-dependent selection of circuits serves different purposes at different levels of visual processing. At the input stage to the striate cortex it contributes to optimize the match between the representations of the two eyes. At a later stage of processing it participates in the development of selective connections between cortical columns and thereby serves to establish neuronal representations for frequently occurring constellations of features. Use-dependent changes of synaptic gain can also be induced in the mature visual cortex. These modifications follow the same rules as those occurring during early development and appear to depend on similar molecular mechanisms. However, in the adult the changes of synaptic gain do not seem to be followed by major rearrangements of connectivity. This suggests developmental alterations in mechanisms responsible for growth, removal and stabilization of synaptic connections. Actually, many of the cellular mechanisms thought to be involved in use-dependent synaptic plasticity change during development but it is still unclear which of them are responsible for the definitive stabilization of functionally confirmed pathways.

Aging↗

PubMed1990

Metabolic effects of reducing rate of glucose ingestion by single bolus versus continuous sipping.

Modifying the rate of absorption has been proposed as a therapeutic principle of specific relevance to diabetes. To demonstrate clearly the metabolic benefits that might result from reducing the rate of nutrient delivery, nine healthy volunteers took 50 g glucose in 700 ml water on two occasions: over 5-10 min (bolus) and at a constant rate over 3.5 h (sipping). Despite similar 4-h blood glucose areas, large reductions were seen in serum insulin (54 +/- 10%, P less than 0.001) and C-peptide (47 +/- 12%, P less than 0.01) areas after sipping, together with lower gastric inhibitory polypeptide and enteroglucagon levels and urinary catecholamine output. There was also prolonged suppression of plasma glucagon, growth hormone, and free-fatty acid (FFA) levels after sipping, whereas these levels rose 3-4 h after the glucose bolus. An intravenous glucose tolerance test at 4 h demonstrated a 48 +/- 10% (P less than 0.01) more rapid decline in blood glucose (Kg) after sipping than after the bolus. Furthermore, FFA and total branched-chain amino acid levels as additional markers of insulin action were lower over this period despite similar absolute levels of insulin and C-peptide. These findings indicate that prolonging the rate of glucose absorption enhances insulin economy and glucose disposal.

Administration, Oral↗

PubMed1990

Disruption of experience-dependent synaptic modifications in striate cortex by infusion of an NMDA receptor antagonist.

To assess the possibility that NMDA receptors play a special role in visual cortical plasticity, the selective antagonist 2-amino-5-phosphonovaleric acid (APV) was continuously infused into the striate cortex of kittens as the visual environment was manipulated during the critical period. The cortex was studied using single-unit recording from sites between 3 and 6 mm from the infusion cannulae. One week of D,L-APV infusion coincident with monocular deprivation or "reverse suture" produced a concentration-dependent increase in the percentage of neurons that (1) lacked normal orientation selectivity and (2) were responsive to stimulation of the deprived eye. These effects outlasted the presence of the drug in the tissue. APV treatment also prevented the acquisition of selectivity and visual responsiveness that normally results from monocular visual experience after dark-rearing. Lasting effects of chronic APV infusion were not observed in adult striate cortex. The effects of APV on kitten striate cortex depended on the presence of the D stereoisomer as infusion of L-APV was without effect. Estimates of extracellular concentration using 3H-APV indicated that significant effects could be obtained with concentrations as low as 20 microM D,L-APV. Recordings from units during infusion indicated that visual responses were reduced by APV. Nonetheless, a normal percentage of visually responsive neurons was found at sites greater than or equal to 3 mm from the infusion cannula. There was no evidence that chronic APV infusion affected the sampling frequency of recorded neurons or disrupted cytoarchitecture at the sites further than 3 mm from the infusion cannula. Taken together, the data indicate that the effects of APV on kitten striate cortex are likely due specifically to the blockade of NMDA receptors. These data are considered in relation to several hypotheses concerning the role of NMDA receptors in the experience-dependent development of striate cortex.

2-Amino-5-phosphonovalerate↗

PubMed1990

Growth hormone suppression and nonproliferative diabetic retinopathy: a preliminary feasibility study.

We used the long acting somatostatin analogue SMS 201-995 in order to examine the feasibility and effect of medical suppression of growth hormone in nonproliferative diabetic retinopathy. Six insulin-dependent diabetic subjects with nonproliferative retinopathy were studied. After eight weeks of SMS 201-995 administration, 24-hour integrated plasma growth hormone concentrations had declined by 47.0 +/- 9.3% of pretreatment values (p less than 0.01), and insulin requirements fell from 40.7 +/- 6.7 units per day to 32.2 +/- 6.9 units per day (p less than 0.01). Plasma levels of somatomedin-C were low before SMS 201-995 (0.5 +/- 0.1 U/ml) and remained unchanged at eight weeks (0.6 +/- 0.1 U/ml; p = ns). During SMS 201-995 administration, best corrected visual acuity improved in both right eyes (53.8 +/- 2.57 to 59.8 +/- 0.7 letters, p less than 0.05) and left eyes (54.8 +/- 2.8 to 61.7 +/- 1.23 letters, p less than 0.03). Fluorescein angiography and stereo fundus photography demonstrated concurrent improvement in retinopathy level in only two subjects. Following cessation of SMS 201-995 treatment, visual acuity returned to pretreatment levels in both right eyes (54.5 +/- 2.4 letters, p = ns compared to baseline) and left eyes (55.8 +/- 2.6 letters, p = ns compared to baseline). These results demonstrate that growth hormone was only partially suppressed by SMS 201-995 in insulin-dependent diabetic subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1989

Nibbling versus gorging: metabolic advantages of increased meal frequency.

We studied the effect of increasing the frequency of meals on serum lipid concentrations and carbohydrate tolerance in normal subjects. Seven men were assigned in random order to two metabolically identical diets. One diet consisted of 17 snacks per day (the nibbling diet), and the other of three meals per day (the three-meal diet); each diet was followed for two weeks. As compared with the three-meal diet, the nibbling diet reduced fasting serum concentrations of total cholesterol, low-density lipoprotein cholesterol, and apolipoprotein B by a mean (+/- SE) of 8.5 +/- 2.5 percent (P less than 0.02), 13.5 +/- 3.4 percent (P less than 0.01), and 15.1 +/- 5.7 percent (P less than 0.05), respectively. Although the mean blood glucose level and serum concentrations of free fatty acids, 3-hydroxybutyrate, and triglyceride were similar during both diets, during the nibbling diet the mean serum insulin level decreased by 27.9 +/- 6.3 percent (P less than 0.01) and the mean 24-hour urinary C-peptide output decreased by 20.2 +/- 5.6 percent (P less than 0.02). In addition, the mean 24-hour urinary cortisol excretion was lower by 17.3 +/- 5.9 percent (P less than 0.05) at the end of the nibbling diet than at the end of the three-meal diet. The blood glucose, serum insulin, and C-peptide responses to a standardized breakfast and the results of an intravenous glucose-tolerance test conducted at the end of each diet were similar. We conclude that in addition to the amount and type of food eaten, the frequency of meals may be an important determinant of fasting serum lipid levels, possibly in relation to changes in insulin secretion.

3-Hydroxybutyric Acid↗

PubMed1989

Blockade of NMDA-receptors prevents ocularity changes in kitten visual cortex after reversed monocular deprivation.

We investigated in the striate cortex of kittens whether the recovery from the effects of monocular deprivation that occurs after reverse occlusion requires activation of N-methyl-D-aspartate (NMDA) receptors. The right eye of 3-4-week-old kittens was closed by lid suture for one week. Subsequently this eye was reopened and the left eyelid sutured closed for another week. During this second week, the NMDA-receptor antagonist, 2-amino-5-phosphonovaleric acid (APV), was infused from an osmotic minipump into the left visual cortex (50 nmol/h), while the right visual cortex was infused only with vehicle solution (saline) as control. At the end of the second week, the ocular dominance of striate cortical neurons was assessed with single unit recording. In the control hemispheres, the large majority of neurons was dominated by the newly opened eye, while in the APV-treated hemispheres most neurons were still dominated by the newly deprived eye. In addition, neurons in the APV-treated hemispheres were less responsive and showed a reduction of orientation tuning. These data confirm that chronic blockade of cortical NMDA-receptors disrupts the disconnection of deprived pathways after monocular deprivation and reduces both responsiveness and orientation selectivity of cortical neurons. In addition they indicate that blockade of NMDA-receptors prevents also vision-dependent recovery of deprived pathways after reverse occlusion.

2-Amino-5-phosphonovalerate↗

PubMed1989

Acetylcholine-induced inhibition in the cat visual cortex is mediated by a GABAergic mechanism.

The influence of iontophoretically applied acetylcholine (ACh) on single-unit activity in the visual cortex was studied in anesthetized cats. The dominant effect consisted of a slow facilitation of neuronal responses to moving light bars. This cholinergic action was sometimes paralleled by a decrease of the cells' selectivity to the direction of stimulus movement. In about one-third of the neurons studied ACh-iontophoresis suppressed maintained and visually driven activity within a few hundred milliseconds from the onset of its application. This effect was antagonized by concurrent iontophoresis of the muscarinic ACh-receptor antagonist scopolamine and the GABAA-receptor antagonist bicuculline methiodide. In 7% of the units studied a fast excitation was elicited by ACh application showing a similar time course as the rapid suppressive effect. It is concluded that ACh-induced inhibition is mediated by an activation of GABAergic interneurons. The role of cholinergic depression and facilitation in cortical information processing is discussed.

Acetylcholine↗

PubMed1989

The development of N-methyl-D-aspartate receptors in cat visual cortex.

During a critical period of early postnatal development, the visual cortex of kittens is susceptible to experience-dependent modifications of neuronal response properties. Recently, the activation of N-methyl-D-aspartate (NMDA) receptors has been identified as an indispensable prerequisite for the induction of such modifications. We therefore investigated developmental changes in the density and distribution of NMDA receptors and questioned whether these showed a relation to the time course of the critical period. We determined the proportion of [3H]glutamate binding sites that were displaced by the NMDA receptor antagonist 2-amino-5-phosphonovalerate (APV) on 10-microns-thick cryostat sections of the primary visual cortex. The overall density of APV-sensitive [3H]glutamate binding sites increased dramatically between the second and the fourth week and stayed at this level throughout the critical period. Towards the end of the critical period, these binding sites decreased and finally reached adult values that were slightly above those of 2-week-old kittens. APV-sensitive binding sites were present in all cortical layers of the age groups investigated. While the general pattern of developmental changes was similar in all layers, slight differences existed in the time course. These observations are compatible with the notion that NMDA receptor activation is required for the expression of use-dependent change of response properties in the kitten visual cortex. Furthermore, they suggest as a possible reason for the decline of malleability towards the end of the critical period the reduction of NMDA receptors.

2-Amino-5-phosphonovalerate↗

PubMed1989

Oscillatory responses in cat visual cortex exhibit inter-columnar synchronization which reflects global stimulus properties.

A fundamental step in visual pattern recognition is the establishment of relations between spatially separate features. Recently, we have shown that neurons in the cat visual cortex have oscillatory responses in the range 40-60 Hz (refs 1, 2) which occur in synchrony for cells in a functional column and are tightly correlated with a local oscillatory field potential. This led us to hypothesize that the synchronization of oscillatory responses of spatially distributed, feature selective cells might be a way to establish relations between features in different parts of the visual field. In support of this hypothesis, we demonstrate here that neurons in spatially separate columns can synchronize their oscillatory responses. The synchronization has, on average, no phase difference, depends on the spatial separation and the orientation preference of the cells and is influenced by global stimulus properties.

Animals↗

PubMed1989

The role of muscarinic acetylcholine receptors in ocular dominance plasticity.

During a critical period of postnatal development neuronal connections in the visual cortex are susceptible to experience-dependent modifications. In normally reared kittens the majority of neurons respond to visual stimulation of either eye. A few days of monocular deprivation, however, are sufficient to render most cortical neurons unresponsive to visual stimuli presented to the deprived eye. Among other factors the cholinergic projection to striate cortex has been identified as having a permissive role in this use-dependent modification of synaptic transmission. In order to analyze further the influence of acetylcholine in cortical plasticity, we tested whether the blockade of muscarinic or nicotinic receptors interfered with ocular dominance plasticity. At four weeks of age kittens had one eyelid sutured closed and osmotic minipumps implanted, which delivered scopolamine (1 nmol/h) or hexamethonium (1 or 10 nmol/h) into the striate cortex of one hemisphere and vehicle solution (saline) into the other. After one week, ocular dominance distributions were determined in area 17 with single unit recording. In the control hemispheres, most neurons became unresponsive to the deprived eye, while in the scopolamine-treated hemispheres most neurons remained binocular. In contrast to the effects of scopolamine, the intracortical infusion of hexamethonium had no effect on ocular dominance plasticity. These results demonstrate that blockade of muscarinic, but not nicotinic receptors renders kitten striate cortex resistant to the effects of monocular deprivation.

Animals↗

PubMed1989

The callosal projection in cat visual cortex as revealed by a combination of retrograde tracing and intracellular injection.

The neuronal composition of callosally projecting cells in cat visual cortex was determined with a combination of retrograde labelling and intracellular injection. Fluorescent tracers were stereotaxically injected into the proximity of the area 17/18 border, corresponding to the representation of the visual vertical meridian. In fixed slice preparations of homotopic regions of the contralateral hemisphere retrogradely labelled cells were filled with Lucifer Yellow. Of more than a hundred injected cells a morphological variety of pyramidal cells, located in cortical layers II-IV and VI, constituted the prevalent cell class in the contralateral projection. A minor proportion of spiny stellate cells was encountered in layer IV. Despite the presence of a contralaterally projecting smooth stellate cell, presumed to be a basket cell, it is concluded that the efferents to contralateral visual cortex predominantly arise from pyramidal and spiny stellate cells. Thus, in agreement with findings from anterograde degeneration studies, the interhemispheric pathway most likely conveys a direct excitatory input to postsynaptic target cells.

Animals↗

PubMed1989

Combining retrograde tracing, intracellular injection, anterograde degeneration and electron microscopy to reveal synaptic links.

Synaptic circuitry was investigated by combining retrograde tracing, intracellular staining, anterograde degeneration and electron microscopy in the same piece of tissue. This methodological procedure was successfully applied to disentangle a disynaptic neuronal chain, which originated in the olfactory bulb, was synaptically relayed in the entorhinal cortex and terminated in the ipsilateral hippocampus. Presumed entorhinal relay cells were retrogradely labelled from their hippocampal termination site by means of a fluorescent tracer. Subsequently, the marked projection neurones were intracellularly injected with Lucifer yellow in fixed slice preparations. Following a simple photo-conversion procedure, dye filled cells were processed for electron microscopy. The origin of presynaptic afferents to identified relay cells was revealed ultrastructurally after lesion induced anterograde degeneration of olfactory mitral cell axons. Due to its reliability, technical simplicity and a high degree of selectivity the new approach is considered an appropriate tool for unravelling neuronal networks.

Animals↗

PubMed1989

Stimulus-specific neuronal oscillations in orientation columns of cat visual cortex.

In areas 17 and 18 of the cat visual cortex the firing probability of neurons, in response to the presentation of optimally aligned light bars within their receptive field, oscillates with a peak frequency near 40 Hz. The neuronal firing pattern is tightly correlated with the phase and amplitude of an oscillatory local field potential recorded through the same electrode. The amplitude of the local field-potential oscillations are maximal in response to stimuli that match the orientation and direction preference of the local cluster of neurons. Single and multiunit recordings from the dorsal lateral geniculate nucleus of the thalamus showed no evidence of oscillations of the neuronal firing probability in the range of 20-70 Hz. The results demonstrate that local neuronal populations in the visual cortex engage in stimulus-specific synchronous oscillations resulting from an intracortical mechanism. The oscillatory responses may provide a general mechanism by which activity patterns in spatially separate regions of the cortex are temporally coordinated.

Animals↗

PubMed1989

Chronic recordings from single sites of kitten striate cortex during experience-dependent modifications of receptive-field properties.

1. With the use of chronically implanted floating microelectrodes, we obtained simultaneous single-unit recordings from multiple sites in the kitten striate cortex and followed experience-dependent modifications of receptive-field properties. For induction of experimental modifications, we used the paradigm of monocular deprivation and reverse occlusion. Kittens were implanted when 4-5 wk old. During the following 2 days, receptive-field properties of the recorded units were determined once under light ketamine anesthesia and repeatedly while the kittens were awake and only lightly restrained. Subsequently, one eye was patched, and the resulting changes in neuronal eye preference were followed by repeated measurements of response properties. For investigation of the effects of reverse occlusion, the deprived eye was opened and the previously open eye closed when the neurons had become unresponsive to the initially deprived eye. Alternatively, kittens were monocularly deprived for 1 wk by lid suture before implantation. The closed eye was then opened, the other eye patched, and the effects of reverse occlusion were studied for up to 1 wk by repeated measurements of receptive-field properties. 2. The earliest effect of monocular deprivation was the disappearance of binocular summation, i.e., binocular responses ceased to be superior over monocular responses. Overt changes of ocular dominance were observed as early as 6 h after the beginning of monocular deprivation. These consisted of a gradual decrease of the excitatory response to deprived eye stimulation and, on occasions, of an additional moderate increase of responses to the normal eye. A complete loss of excitatory responses to deprived-eye stimulation was seen as early as 12 h after occlusion. In numerous cells, however, stimulation of the deprived eye continued to evoke inhibitory responses even after excitatory responses had vanished completely. During this shift in ocularity, neurons preserved their orientation and direction selectively. 3. The minimal time required for the manifestation of ocular dominance changes was similar regardless of whether the animals were stimulated continuously or were asleep part of the time, suggesting the existence of an experience-independent consolidation period for ocular dominance changes. 4. The first change after reverse occlusion was a reduction of the response to the newly deprived eye. The time course of this inactivation was similar to that observed after initial deprivation, whereas the recovery of responses to the previously deprived eye had a considerably slower time course.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1989

The effects of SMS 201-995 (sandostatin) on metabolic profiles in insulin-dependent diabetes mellitus.

GH has been implicated in the pathophysiology of various acute and chronic complications of diabetes mellitus. As a consequence, there has been a great deal of interest in developing methods for suppressing GH secretion in diabetes. SMS 201-995 is a long-acting somatostatin analog which inhibits the secretion of numerous hormones, including GH. To determine the metabolic and hormonal responses to SMS 201-995 independent of endogenous insulin suppression, we studied six patients with insulin-dependent diabetes mellitus while they received 150 micrograms SMS 201-995, sc, daily for an 8-week period. This treatment resulted in no change in 24-h glucose profiles, although the mean insulin dose decreased by 19%, while hemoglobin A1c decreased significantly (0.084 +/- 0.023 to 0.067 +/- 0.011, P = 0.04). The 24-h profiles of blood lactate, plasma free insulin, glucagon, FFA, blood glycerol, and beta-hydroxybutyrate were unchanged, whereas that of blood alanine increased significantly (7.8 +/- 0.4 to 10.6 +/- 0.9 mmol/L.h; P = 0.01). GH secretion declined in five of the six patients; the mean values before and during SMS 201-995 treatment were 102 +/- 23 and 68 +/- 12 micrograms/L.h, respectively (P = NS), for the six patients. [In the five patients in whom GH secretion declined, the mean values before and during SMS 201-995 treatment were 115 +/- 23 and 63 +/- 14 micrograms/L.h, respectively (P = 0.01).] These results suggest that SMS 201-995 may be administered to patients with insulin-dependent diabetes mellitus without a deleterious effect on metabolic control.

Adult↗

PubMed1988

Pharmacological induction of use-dependent receptive field modifications in the visual cortex.

Lasting modifications of the receptive fields of neurons in the visual cortex can be induced by pairing visual stimuli with iontophoretic application of the neuromodulators acetylcholine and noradrenaline or the excitatory amino acids N-methyl-D-aspartate (NMDA) and L-glutamate. The modifications are obtained in less than 1 hour and persist for more than 40 minutes. Thus, acetylcholine and norepinephrine have a permissive role in use-dependent neuronal plasticity. These results support the notion of a postsynaptic threshold for neuronal malleability that differs from that of sodium-dependent action potentials.

Acetylcholine↗

PubMed1988

Light and electron microscopic immunocytochemical localization of parvalbumin in the dorsal lateral geniculate nucleus of the cat: evidence for coexistence with GABA.

The coexistence in individual neurons of parvalbumin and gamma-aminobutyric acid (GABA) was studied in the dorsal lateral geniculate nucleus (dLGN) of the cat using pre- and postembedding immunocytochemical methods. PV(+) cell bodies and processes were found in the perigeniculate nucleus (PGN) and throughout all laminae of the dLGN. PV(+) neurons were relatively small and had circular to fusiform shapes. Electron microscopy revealed PV(+) reaction product within the perikarya, axons, and dendrites of labeled cells. It was associated preferentially with microtubules, postsynaptic densities, and intracellular membranes. PV(+) presynaptic boutons were identified on the basis of their synaptic relations and ultrastructure as retinal terminals (RLP) and as profiles originating from inhibitory interneurons (F1 and F2). Immunopositive somata and dendrites received asymmetric synaptic contacts from labeled RLP and non-identified, non-immunoreactive synaptic boutons. Moreover, PV(+) dendrites were postsynaptic to labeled F profiles. In the PGN all neurons were both PV(+) and GABA-immunoreactive and in the dLGN the vast majority of PV(+) neurons showed GABA-immunoreactivity. It is suggested that the high incidence of PV in GABAergic neurons is related to the particular activation patterns of these neurons and the resulting demand for calcium buffer systems.

Animals↗