Biomedical subjects

K Langley

Publications and source records attributed to K Langley.

At least 37 records · Page 2Linked to original sources

PubMed2000

7beta-hydroxysterol is cytotoxic to neonatal rat astrocytes in primary culture when cAMP levels are increased.

We have shown previously that 7beta-hydroxycholesterol (7betaOHCH) and 7beta-hydroxycholesteryl-3-oleate (7betaOHCH- 3-OL) are potent inhibitors of lesion-induced astrogliosis in the rat cortex or spinal cord; these substances reduce reactive astrocyte proliferation and hypertrophy. In this study, we employed cultured newborn rat astrocytes with increased cAMP levels as an in vitro model of reactive astrocytes. Treatment with either dibutyryl-cAMP (dbcAMP) or isoproterenol resulted in morphologic differentiation of astrocytes which became fibrous. Concomitant incubation with 30 microM 7betaOHCH and dbcAMP (or isoproterenol) provoked the cells to retract and was cytotoxic. When the beta-adrenergic receptor-mediated cAMP increase was abolished by propranolol, the 7betaOHCH cytotoxicity was inhibited. Immunocytochemical labelling for glial fibrillary acidic protein (GFAP) and beta-tubulin and electron microscopy suggested that intermediate filament and microtubular organizations were modified by 7betaOHCH. Analysis of the activity of cAMP-dependent protein kinase (PKA) in astrocytes treated with dbcAMP and 7betaOHCH showed a rapid and marked inhibition of the phosphotransferase activity which lasted for 24 hr. We suggest that this culture system provides an experimental system to study the molecular mechanisms involved in the effect of oxysterols on astrocytic hypertrophy. The cytotoxicity of 7betaOHCH seems to be mediated by inhibition of PKA, which phosphorylates intermediate filaments and the transcription factor cyclic AMP responsive element binding.

Adrenergic beta-Agonists↗

PubMed2000

SNAP-25 regulation during adrenal gland development: comparison with differentiation markers and other SNAREs.

Synaptosomal-associated protein of 25 kDa (SNAP-25) is one of a limited number of soluble N-ethylmaleimide-sensitive fusion attachment protein receptors (SNAREs) that play a major role in membrane docking of synaptic vesicles and secretory granules during regulated exocytosis. We have previously shown that SNAP-25 levels differ between noradrenergic and adrenergic chromaffin cell populations of the adult adrenal gland. We examine SNAP-25 expression by immunofluoresence in cells of the sympathoadrenal lineage in the rat during late embryonic and postnatal development. In parallel, tyrosine hydroxylase was used to identify sympathoadrenal cells, phenylethanolamine N-methyltransferase to distinguish adrenergic from noradrenergic chromaffin cells, and chromogranin A to define the presence of secretory granules. In addition, SNAP-25 protein and mRNA levels were followed in adrenal gland extracts by immunoblotting and reverse transcription-polymerase chain reaction (RT-PCR). Protein levels were compared with those of other molecules also implicated in organelle trafficking, including syntaxin 1 and vesicle-associated membrane protein (VAMP-2) and the nonneuronal analogues SNAP-23 and cellubrevin. This study provides evidence that SNAP-25 is expressed early during development in sympathoadrenal neurons and migrating cells. It is detected in intra-adrenal chromoblasts as soon as they enter the adrenal primordium. Its differential expression between catecholamine chromaffin cell phenotypes is already evident from the 17th embryonic day, future noradrenergic cells appearing to express higher levels than adrenergic cells. The granule maturation marker chromogranin A is expressed in chromaffin cells later than SNAP-25. Both SNAP-25 protein and mRNA increased rapidly in the adrenal gland in the perinatal period to peak during the first postnatal week, after which levels dropped dramatically to adult values. In contrast, levels of both syntaxin and SNAP-23 appeared to remain fairly constant throughout adrenal gland development. VAMP-2 expression increased gradually around birth to reach maximal levels during the first two postnatal weeks, and then decreased slightly. Cellubrevin levels also appeared to increase gradually until adult values were attained by the end of the second postnatal week. The threefold increase of SNAP-25 mRNA shortly after birth compared to the low adult levels suggests that during this period SNAP-25 is implicated in additional functions than regulated secretion, possibly associated with cellular growth or maturation.

Adrenal Glands↗

PubMed2000

A model of motion adaptation and motion after-effects based upon principal component regression.

A computational model to help explain effects of adaptation to moving signals is compared with established energy (linear regression) models of motion detection. The proposed model assumes that processed image signals are subject to error in both dimensions of space and time. This assumption constrains models of motion perception to be based upon principal component regression rather than linear regression. It is shown that response suppression of model complex cell neurons that input into the model may account for (1) increases in perceived speed after adaptation to static patterns and testing with slowly moving patterns, (2) significant increases in perceived speed after adaptation to patterns moving at a medium speed and testing at high speed, and (3) decreases in perceived speed in the opponent direction to a quickly moving adapting signal. Neither of predictions (2) or (3) are general features of established accounts of motion detection by visual processes based upon linear regression. Comparisons of the proposed model's speed transfer function with existing psychophysical data suggests that the visual system processes motion signals with the tacit assumption that image measurements are subject to error in both space and time.

Animals↗

PubMed2000

Recursive implementations of temporal filters for image motion computation.

Efficient algorithms for image motion computation are important for computer vision applications and the modelling of biological vision systems. Intensity-based image motion computation proceeds in two stages: the convolution of linear spatiotemporal filter kernels with the image sequence, followed by the non-linear combination of the filter outputs. If the spatiotemporal extent of the filter kernels is large, then the convolution stage can be very intensive computationally. One effective means of reducing the storage required and computation involved in implementing the temporal convolutions is the introduction of recursive filtering. Non-recursive methods require the number of frames of the image sequence stored at any given time to be equal to the temporal extent of the slowest temporal filter. In contrast, recursive methods encode recent stimulus history implicitly in the values of a small number of variables updated through a series of feedback equations. Recursive filtering reduces the number of values stored in memory during convolution and the number of mathematical operations involved in computing the filters' outputs. This paper extends previous recursive implementations of gradient- and correlation-based motion analysis algorithms [Fleet DJ, Langley K (1995) IEEE PAMI 17: 61-67; Clifford CWG, Ibbotson MR, Langley K (1997) Vis Neurosci 14: 741-749], describing a recursive implementation of causal band-pass temporal filters suitable for use in energy- and phase-based algorithms for image motion computation. It is shown that the filters' temporal frequency tuning curves fit psychophysical estimates of the temporal properties of human visual filters.

Algorithms↗

PubMed2000

Phospholipid composition in late infantile neuronal ceroid lipofuscinosis.

BACKGROUND: Neuronal ceroid lipofuscinosis (NCL) is a relatively common group of inherited neurodegenerative disorders characterised by the accumulation of autofluorescent lipopigments (ceroid) similar to lipofuscin. Because of this property, studies have concentrated on fatty acid metabolism and lipid peroxidation. METHODS: In the present study, the fatty acid composition of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) and the molecular species compositions of diacylglycerophosphocholine (diacyl GPC), diacylglycerophosphoethanolamine (diacyl GPE) and alkenylacyl GPE (plasmalogens) were investigated in cultured skin fibroblasts from three patients with a confirmed diagnosis of the late infantile form of the disease (LINCL, CLN2) and three healthy age-matched controls. RESULTS: Relatively minor differences in the fatty acid compositions of PC and PE were observed between patients and controls. However, dimethyl acetals of plasmalogens were found to be 40% higher in the patients compared to in the controls. Control and LINCL fibroblasts displayed only slight differences in the molecular compositions of diacyl GPE and diacyl GPC. In contrast, compared with normal cells, LINCL fibroblasts had higher levels of alkenylacyl GPE species containing both 18 : 1 and polyunsaturated fatty acids, but lower levels of species with 16 : 0 or 18 : 0 in the sn-1 position. CONCLUSION: The molecular composition of PC and PE subclasses in skin fibroblasts of healthy subjects and patients suffering from LINCL is here described for the first time. While few differences are noticeable in the fatty acid composition of PC and PE and the molecular species distribution of diacylGPC and diacylGPE, the alkenylacyl GPE (or ethanolamine plasmalogens) were found to differ significantly between patients and healthy controls.

Cells, Cultured↗

PubMed1999

Cultured glial cells express the SNAP-25 analogue SNAP-23.

Astrocytes release glutamate and aspartate in response to elevated intracellular calcium levels, and it has been proposed that this occurs by a vesicular release mechanism, in which SNARE proteins are implicated. Although syntaxin, synaptobrevin, and cellubrevin have been shown to be expressed by cultured astrocytes, SNAP-25 has not been detected. By using immunocytochemical, immunoblotting, and polymerase chain reaction techniques, the present study demonstrates that SNAP-23, an analogue of SNAP-25, is expressed by astrocytes both in culture and in rat cerebellum. These findings provide additional evidence that astrocytes release excitatory amino acids by a vesicular mechanism involving SNARE proteins. SNAP-23 and also syntaxin 1 and cellubrevin were found to be expressed in glial precursor cells, oligodendrocytes, and microglia. These data suggest that the t-SNAREs SNAP-23 and syntaxin 1 and the v-SNARE cellubrevin participate in general membrane insertion mechanisms involved in diverse glial cell functions such as secretion, phagocytosis, and myelinogenesis.

Animals↗

PubMed1999

Molecular markers of sympathoadrenal cells.

Cells constituting the sympathoadrenal (SA) cell lineage originate from the neural crest and acquire a catecholaminergic fate following migration to the dorsal aorta. Subsequently, SA cells migrate to sites widely dispersed throughout the body. In addition to endocrine chromaffin and "small intensely fluorescent" cells in adrenal glands and in extra-adrenal tissues such as the paraganglia, this lineage also includes neurones located in sympathetic ganglia and in the adrenal gland. It is widely assumed that these cells are all derived from the same precursors, which then differentiate along divergent pathways in response to different external stimuli. During embryonic differentiation, SA cells lose some of their early traits and acquire other distinguishing features. To help understand how the lineage diverges in terms of phenotype and function, this article examines the cellular expression of a variety of "marker" proteins that characterize the individuals of the lineage. In particular, differences between adrenal medullary adrenergic and noradrenergic chromaffin cells in the expression of proteins, such as the neural adhesion molecule L1, the growth-associated protein GAP-43 and molecules involved in the secretory process, are emphasized. Factors that might differentially regulate such molecular markers in these cells are discussed.

Adrenal Glands↗

PubMed1999

Computational models of coherent and transparent plaid motion.

The perceived motion of two added sinusoidal gratings of similar amplitude and spatial frequency but different orientations is often coherent. However, when either relative grating contrast or frequency are varied, perception may transform to a motion transparency. For plaids, both multiplicative and additive transparent percepts are reported. To explain perception, several computational models of motion transparency are proposed. The most general model considered is, however, a quadratic form with five unknowns. To stabilize the transparent model, additional constraints are introduced so that two velocities may be detected from the motion of plaid patterns. It is shown how this model may be realised by a two-layer (linear) feedforward network and how network learning paradigms may be used to explain some facets of visual perception. To describe the motion of plaid patterns there is an ambiguity because computational models of both coherent and transparent motion may be used to detect image velocity. In view of this competition between models, the issue of model selection is addressed; especially for cases where two or more models fit the image measurements without a residual error. The computational approach that is proposed affords one explanation why perception selects transparency in favour of coherence for plaid patterns by adjustments of relative grating contrast and frequency.

Humans↗

PubMed1999

Stereopsis from contrast envelopes.

We report two experiments concerning the site of the principal nonlinearity in second-order stereopsis. The first exploits the asymmetry in perceiving transparency with second-order stimuli found by Langley et al. (1998) (Proceedings of the Royal Society of London B, 265, 1837-1845) i.e. the product of a positive-valued contrast envelope and a mean-zero carrier grating can be seen transparently only when the disparities are consistent with the envelope appearing in front of the carrier. We measured the energy at the envelope frequencies that must be added in order to negate this asymmetry. We report that this amplitude can be predicted from the envelope sidebands and not from the magnitude of compressive pre-cortical nonlinearities measured by other researchers. In the second experiment, contrast threshold elevations were measured for the discrimination of envelope disparities following adaptation to sinusoidal gratings. It is reported that perception of the envelope's depth was affected most when the adapting grating was similar (in orientation and frequency) to the carrier, rather than to the contrast envelope. These results suggest that the principal nonlinearity in second-order stereopsis is cortical, occurring after orientation- and frequency-selective linear filtering.

Adaptation, Ocular↗

PubMed1999

Physiological aspects of exocytosis in chromaffin cells of the adrenal medulla.

The adrenal medulla is composed principally of groups of adrenergic and noradrenergic chromaffin cells, with minor populations of small intensely fluorescent cells and ganglionic neurones. Different molecular stimuli evoke distinct secretory events in the gland, involving the release of either adrenaline or noradrenaline together with various neuroactive peptides. The nature of the secretory response can be controlled at a central level or regulated locally within the gland. Specific innervation patterns to the different types of chromaffin cell have been implicated in central regulatory mechanisms, while several explanations for regulating secretion locally have been proposed. The differential distribution of various types of receptors between cell phenotypes, such as muscarinic or nicotinic acetylcholine receptors, histamine receptors, angiotensin receptors and different classes of opiate receptors between the two principal chromaffin cell populations could be involved in local control. In addition exocytosis parameters could be modulated differently in adrenergic and noradrenergic cells by phenotype-specific mechanisms, possibly involving molecules like Growth Associated Protein-43, Synaptosomal Associated Protein-25 isoforms or the p11 annexin subunit. The distribution of the various types of calcium channels is also known to vary between chromaffin cell subtypes. This short review examines possible ways in which specific innervation patterns in the adrenal gland could be programmed and discusses exocytosis mechanisms that could differ between chromaffin cell phenotypes. Data reviewed here suggest that the adrenal medulla should no longer be viewed as a homogeneous entity but as consisting of an ensemble of individual cell subpopulations each with a distinct secretory response that could in part reflect its local history.

Adrenal Medulla↗

PubMed1999

Differential expression of SNAP-25 isoforms and SNAP-23 in the adrenal gland.

In the rat adrenal gland, we previously observed that SNAP-25 is not restricted to the plasmalemma in noradrenergic cells as it is in adrenergic cells, and hypothesized that SNAP-25 isoform expression is different in the two phenotypes. Expression of SNAP-25 isoforms and SNAP-23 was examined by immunoblotting, immunofluorescence, and RT-PCR. Amplifications of SNAP-25 mRNAs were combined with Southern hybridization, restriction enzyme analysis, and sequencing of cloned PCR products to compare SNAP-25 isoform expression in rat and bovine adrenal glands. SNAP-25 and SNAP-23 mRNA and protein are expressed in the glands; SNAP-23 is enriched in the adrenal cortex, whereas SNAP-25 is restricted to the adrenal medulla. Furthermore, high levels of SNAP-25 and low levels of SNAP-23 are observed in the PC12 cells, whereas both SNAP-25 and SNAP-23 are expressed in adrenal medullary cultures. In all extracts, the SNAP-23 mRNA corresponded to SNAP-23a. SNAP-25a is the major form expressed in rat adrenal glands (75%), as it is in PC12 cells (80%), but both SNAP-25a and SNAP-25b (40% vs. 60%) are expressed in bovine adrenal medulla in situ and in culture. In addition, an enriched population of adrenergic cells (93%) expressed a higher level of SNAP-25b (70%), suggesting that this isoform may not be restricted to fast neurotransmission.

Adrenal Glands↗

PubMed1999

Clustering of activating mutations in c-KIT's juxtamembrane coding region in canine mast cell neoplasms.

The proto-oncogene c-KIT encodes a growth factor receptor, KIT, with ligand-dependent tyrosine kinase activity that is expressed by several cell types including mast cells. c-KIT juxtamembrane coding region mutations causing constitutive activation of KIT are capable of transforming cell lines and have been identified in a human mast cell line and in situ in human gastrointestinal stromal tumors, but have not been demonstrated in situ in neoplastic mast cells from any species. To determine whether c-KIT juxtamembrane mutations occur in the development of mast cell neoplasms, we examined canine mastocytomas, which are among the most common tumors of dogs and which often behave in a malignant fashion, unlike human solitary mastocytomas. Sequencing of c-KIT cDNA generated from tumor tissues removed from seven dogs revealed that three of the tumors contained a total of four mutations in an intracellular juxtamembrane coding region that is completely conserved among vertebrates. In addition, two mutations were found in three mast cell lines derived from two additional dogs. One mutation from one line matched that found in situ in one of the tumors. The second was found in two lines derived from one dog at different times, indicating that the mutation was present in situ in the animal. All five mutations cause high spontaneous tyrosine phosphorylation of KIT. Our study provides in situ evidence that activating c-KIT juxtamembrane mutations are present in, and may therefore contribute to, the pathogenesis of mast cell neoplasia. Our data also suggest an inhibitory role for the KIT juxtamembrane region in controlling the receptor kinase activity.

Amino Acid Sequence↗

PubMed1998

Linear and nonlinear transparencies in binocular vision.

When the product of a vertical square-wave grating (contrast envelope) and a horizontal sinusoidal grating (carrier) are viewed binocularly with different disparity cues they can be perceived transparently at different depths. We found, however, that the transparency was asymmetric; it only occurred when the envelope was perceived to be the overlaying surface. When the same two signals were added, the percept of transparency was symmetrical; either signal could be seen in front of or behind the other at different depths. Differences between these multiplicative and additive signal combinations were examined in two experiments. In one, we measured disparity thresholds for transparency as a function of the spatial frequency of the envelope. In the other, we measured disparity discrimination thresholds. In both experiments the thresholds for the multiplicative condition, unlike the additive condition, showed distinct minima at low envelope frequencies. The different sensitivity curves found for multiplicative and additive signal combinations suggest that different processes mediated the disparity signal. The data are consistent with a two-channel model of binocular matching, with multiple depth cues represented at single retinal locations.

Depth Perception↗

PubMed1998

Plaid slant and inclination thresholds can be predicted from components.

We investigated whether stereoscopic slant and inclination thresholds for surfaces defined by two component plaids could be predicted from the interocular differences in their individual component gratings. Thresholds were measured for binocular images defined by single sinusoidal gratings and two component plaids. In both cases thresholds showed a marked dependence on component orientation. For absolute component orientations greater than 45 deg we found that inclination thresholds were smaller than slant thresholds. However, for absolute component orientations less than 45 deg, we found a reversal: slant thresholds were smaller than inclination thresholds. We considered three models that might account for these data. One assumed that thresholds stemmed from interocular position differences of corresponding image points. The other two assumed a combination of position, orientation and/or spatial-frequency differences. The best fits were obtained from those models that explicitly represented orientation differences. From the model combining orientation and spatial-frequency differences, we estimated the relative cue sensitivity to be 1.7:1, respectively. For plaids, we found that thresholds obtained from the individual components could be used to predict thresholds for plaids, even though an additional disparity cue from the contrast beat was available.

Depth Perception↗

PubMed1997

Chymase cleavage of stem cell factor yields a bioactive, soluble product.

Stem cell factor (SCF) is produced by stromal cells as a membrane-bound molecule, which may be proteolytically cleaved at a site close to the membrane to produce a soluble bioactive form. The proteases producing this cleavage are unknown. In this study, we demonstrate that human mast cell chymase, a chymotrypsin-like protease, cleaves SCF at a novel site. Cleavage is at the peptide bond between Phe-158 and Met-159, which are encoded by exon 6 of the SCF gene. This cleavage results in a soluble bioactive product that is 7 amino acids shorter at the C terminus than previously identified soluble SCF. This research shows the identification of a physiologically relevant enzyme that specifically cleaves SCF. Because mast cells express the KIT protein, the receptor for SCF, and respond to SCF by proliferation and degranulation, this observation identifies a possible feedback loop in which chymase released from mast cell secretory granules may solubilize SCF bound to the membrane of surrounding stromal cells. The liberated soluble SCF may in turn stimulate mast cell proliferation and differentiated functions; this loop could contribute to abnormal accumulations of mast cells in the skin and hyperpigmentation at sites of chronic cutaneous inflammation.

Binding Sites↗

PubMed1997

Are exocytosis mechanisms neurotransmitter specific?

Neurotransmission is a multistage regulated process in which a variety of active molecules contained in vesicles are liberated in response to specific stimuli from different types of neurone or related cells. This includes the release of fast neurotransmitters such as amino acids and acetylcholine from central and peripheral synapses, but also that of relatively slow-acting polypeptides from central and peripheral neurones or neuroendocrine cells. Considerable progress has been made over recent years in the understanding at a molecular level of the mechanism of regulated exocytosis, a crucial phase in this phenomenon. The currently proposed overall mechanism, which incorporates the "SNARE" hypothesis for vesicle-membrane docking and fusion, is based on data from experimental models ranging from brain synaptosomes to mast cells. Since the kinetics of the models studied and the physiological effects of the neurotransmitters implicated vary so much, it is pertinent to question whether a general mechanism can be proposed from such experimental data. This review examines known differences in putative exocytotic mechanisms for the various systems studied and attempts to relate these to the nature of the active substances released. Differences exist in each step of the exocytosis process and include the channel through which Ca2+ enters to trigger it or the internal Ca2- source, the type of vesicle in which the transmitter is packaged, the way vesicles are translocated to the surface membrane or how they dock and fuse with it. Major differences have been reported in release mechanisms of different types of vesicle, but minor differences also exist within the same vesicle class. Thus small synaptic vesicles and large dense core vesicles are translocated by distinct processes and the Ca2+ channels, Ca2+ sensors and docking proteins involved in other steps are not identical in all neuronal phenotypes. It may be concluded that each of these differences has evolved to accommodate the different physiological requirements of the neuromodulator released.

Animals↗

PubMed1996

Psychophysics of motion adaptation parallels insect electrophysiology.

We investigate the form and time course of motion adaptation, comparing the psychophysical performance of human subjects with existing electrophysiological data on insect vision. In the H1 neuron of the fly, the response to a maintained motion stimulus is known to decrease over time while sensitivity to variations in speed around the maintained level increases. This behaviour can be modelled by modifying a correlation-based motion detector to include adaptable temporal filters (Fig. 1). We find that the form and time course of sensitivity changes in human motion perception are comparable to fly vision. We propose that, in both cases, adaptation serves to improve the transmission of novel motion information along the visual pathways at the expense of maintaining an accurate representation of the unchanging components of the stimulus.

Adaptation, Physiological↗

PubMed1996

Structure-function relationships of stem cell factor: an analysis based on a series of human-murine stem cell factor chimera and the mapping of a neutralizing monoclonal antibody.

Although much is now known about the biological properties of the c-kit receptor and its ligand, stem cell factor (SCF), little is known of the structural basis for the binding and function of this hematopoietic cytokine. By analyzing the activities of chimeric interspecies and homologue muteins and epitope mapping of a monoclonal antibody (MoAb) to the human protein, we have found that three distinct regions of SCF are essential for full biological function. Homologue and interspecies swapping of polypeptide sequences between the amino terminus and G35, between L79 and N97, and between R121 and D128 reduced or eliminated the ability of the chimera to act in synergy with murine granulocyte-macrophage colony-stimulating factor (GM-CSF) to promote hematopoietic colony formation. Moreover, a nonconformation-dependent MoAb that neutralizes human, but not murine SCF, was found to bind to residues within the L79-N97 segment of the human homologue. As these three regions localize to the putative first, third, and fourth helices of the protein, findings remarkably similar to previous studies of cytokines as diverse as growth hormone, GM-CSF, and interleukin (IL)-4, our results suggest that cytokines of multiple classes share a common functional organization.

Animals↗