Biomedical subjects

G J Michael

Publications and source records attributed to G J Michael.

At least 37 records · Page 2Linked to original sources

PubMed1999

Axotomy results in major changes in BDNF expression by dorsal root ganglion cells: BDNF expression in large trkB and trkC cells, in pericellular baskets, and in projections to deep dorsal horn and dorsal column nuclei.

Brain derived neurotrophic factor (BDNF) is normally expressed by a small number of predominantly trkA-expressing dorsal root ganglion cells. Using immunocytochemistry and in situ hybridization, we have examined the effect of sciatic nerve section on the expression of BDNF in the adult rat. Following axotomy there was a long lasting (4-week) increase in BDNF mRNA and protein in large-diameter, trkB- and trkC-expressing dorsal root ganglion cells. By 2 days postaxotomy, expression of BDNF mRNA had increased from 2% of trkB cells to 50%, and from 18% of trkC cells to 56%. In contrast, BDNF expression in most trkA cells was unchanged, although was increased in the small population of medium- and large-sized trkA cells. Following axotomy, BDNF-immunoreactive terminals appeared in the central axonal projections of large-diameter cells, including the deep dorsal horn and gracile nucleus. Neuropeptide Y was also upregulated following axotomy and was coexpressed with BDNF in the cell bodies and central terminals of the large cells. Ultrastructural analysis in lamina IV of the spinal cord revealed that BDNF terminals in these central projections establish synaptic contacts. Immunoreactivity at 4 weeks was also observed in pericellular baskets that contained calcitonin gene-related peptide (CGRP) and surrounded trkA- and trkB-expressing cells in L4 and L5 lumbar ganglia. These baskets are likely to arise from local, highly immunoreactive, BDNF/CGRP/trkA-expressing cells. Our results identify several novel targets for BDNF and imply that it acts locally in both autocrine and paracrine modes, as well as centrally in a synaptic mode, to modulate the response of somatosensory pathways in nerve injury.

Afferent Pathways↗

PubMed1999

Simulation studies of optimum energies for DXA: dependence on tissue type, patient size and dose model.

Dual-energy x-ray absorptiometry (DXA) is a well established technique for measuring bone mineral density (BMD). However, in recent years DXA is increasingly being used to measure body composition in terms of fat and fat-free mass. DXA scanners must also determine the soft tissue baseline value from soft-tissue-only regions adjacent to bone. The aim of this work is to determine, using computer simulations, the optimum x-ray energies for: a number of dose models, different tissues, i.e. bone mineral, average soft tissue, lean soft tissue and fat; and a range of anatomical sites and patient sizes. Three models for patient dose were evaluated: total beam energy, entrance exposure and absorbed dose calculated by Monte Carlo modelling. A range of tissue compositions and thicknesses were chosen to cover typical patient variations for the three sites: femoral neck, PA spine and lateral spine. In this work, the optimisation of the energies is based on: (1) the uncertainty that arises from the quantum statistical nature of the number of x-rays recorded by the detector, and (2) the radiation dose received by the patient. This study has deliberately not considered other parameters such as detector response, electronic noise, x-ray tube heat load etc, because these are technology dependent parameters, not ones that are inherent to the measuring technique. Optimisation of the energies is achieved by minimisation of the product of variance of density measurement and dose which is independent of the absolute intensities of the x-ray beams. The results obtained indicate that if solving for bone density, then Elow in the range 34 to 42 keV, Ehigh in the range 100 to 200 keV and incident intensity ratio (low energy/high energy) in the range 3 to 10 is a reasonable compromise for the normal range of patient sizes. The choice of energies is complicated by the fact that the DXA unit must also solve for fat and lean soft tissue in soft-tissue-only regions adjacent to the bone. In this case the ranges of energies suggested above are acceptable only for the average phantom. In extreme cases the variance-dose product can be up to 50 times higher than for the ideal energies.

Absorptiometry, Photon↗

PubMed1998

Growth associated protein 43 (GAP-43) mRNA is upregulated in the rat superior cervical ganglia after preganglionic transection.

Growth-associated protein 43 (GAP-43) is a growth-associated protein which is synthesised in high amounts in neurons during neuronal outgrowth. In a previous study we have shown that GAP-43 immunoreactivity is increased in neurons in superior cervical ganglia (SCG) and in nerve terminals in the irides after preganglionic denervation. We have now examined changes in GAP-43 mRNA using in situ hybridisation. GAP-43 mRNA was seen to be constitutively expressed by principal neurons of the rat superior cervical ganglion. Expression was increased further by section of the cervical sympathetic trunk, reaching a maximum (increased by about 30%) 3 days after decentralisation. The increased GAP-43 protein seen after decentralisation thus appears to be due to an upregulation of GAP-43 mRNA in the adrenergic neurons. The results imply that GAP-43 expression in the SCG is under presynaptic control, acting at least partly by control of mRNA levels.

Animals↗

PubMed1998

A distinct subgroup of small DRG cells express GDNF receptor components and GDNF is protective for these neurons after nerve injury.

Several lines of evidence suggest that neurotrophin administration may be of some therapeutic benefit in the treatment of peripheral neuropathy. However, a third of sensory neurons do not express receptors for the neurotrophins. These neurons are of small diameter and can be identified by the binding of the lectin IB4 and the expression of the enzyme thiamine monophosphatase (TMP). Here we show that these neurons express the receptor components for glial-derived neurotrophic factor (GDNF) signaling (RET, GFRalpha-1, and GFRalpha-2). In lumbar dorsal root ganglia, virtually all IB4-labeled cells express RET mRNA, and the majority of these cells (79%) also express GFRalpha-1, GFRalpha-2, or GFRalpha-1 plus GFRalpha-2. GDNF, but not nerve growth factor (NGF), can prevent several axotomy-induced changes in these neurons, including the downregulation of IB4 binding, TMP activity, and somatostatin expression. GDNF also prevents the slowing of conduction velocity that normally occurs after axotomy in a population of small diameter DRG cells and the A-fiber sprouting into lamina II of the dorsal horn. GDNF therefore may be useful in the treatment of peripheral neuropathies and may protect peripheral neurons that are refractory to neurotrophin treatment.

Age Factors↗

PubMed1998

Monte Carlo modelling of an extended DXA technique.

The precision achieved in measuring bone mineral density (BMD) by commercial dual-energy x-ray absorptiometry (DXA) machines is typically better than 1%, but accuracy is considerably worse. Errors, due to inhomogeneous distributions of fat, of up to 10% have been reported. These errors arise because the DXA technique assumes a two-component model for the human body, i.e. bone mineral and soft tissue. This paper describes an extended DXA technique that uses a three-component model of human tissue and significantly reduces errors due to inhomogeneous fat distribution. In addition to two x-ray transmission measurements, a measurement of the path length of the x-ray beam within the patient is required. This provides a third equation, i.e. T = ts + tb + tf where T, ts, tb and tf are the total, lean soft tissue, bone mineral and fatty tissue thicknesses respectively. Monte Carlo modelling was undertaken to make a comparison of the standard and extended DXA techniques in the presence of inhomogeneous fat distribution. Two geometries of varying complexity were simulated. In each case the extended DXA technique produced BMD measurements that were independent of soft tissue composition whereas the standard technique produced BMD measurements that were strongly dependent on soft tissue composition. For example, in one case, the gradients of the plots of BMD versus fractional fat content were for standard DXA (-0.183+/-0.037) g cm(-2) and for extended DXA (0.027+/-0.044) g cm(-2). In all cases the extended DXA method produced more accurate but less precise results than the standard DXA technique.

Absorptiometry, Photon↗

PubMed1998

Effect of nerve growth factor treatment on p75NTR gene expression in lumbar dorsal root ganglia of streptozocin-induced diabetic rats.

Previous work shows that gene expression for p75NTR in lumbar dorsal root ganglia (DRG) is deficient in streptozocin-induced diabetic rats, while expression of trkA protein is unaffected. This is the first study on the effect of diabetes on immunohistochemical staining and axonal transport of p75NTR in sensory neurons. We also investigated the novel effect of nerve growth factor (NGF) treatment on the levels of mRNA and protein of the NGF receptors, trkA and p75NTR, in normal and diabetic rats. Immunohistochemical staining for p75NTR was significantly reduced in DRG of 12-week streptozocin-induced diabetic rats, confirming the previous work. Anterograde and retrograde axonal transport of p75NTR within the sciatic nerve was similarly decreased in diabetic rats, while levels of transport of trkA were unaffected. Treatment with systemic NGF reversed the diabetes-induced deficit in p75NTR transcripts and protein within the DRG and left expression levels of trkA unchanged. We propose that in sensory neurons of diabetic rats, the ability to capture and retrogradely transport NGF may be impaired because of suboptimal production of p75NTR receptors and that NGF therapy may overcome this deficiency.

Animals↗

PubMed1997

Nerve growth factor treatment increases brain-derived neurotrophic factor selectively in TrkA-expressing dorsal root ganglion cells and in their central terminations within the spinal cord.

Using immunocytochemistry and in situ hybridization, we have examined the expression of brain-derived neurotrophic factor (BDNF) and of neurotrophin receptors in dorsal root ganglion cells. In the adult rat, BDNF mRNA and protein were found mainly in the subpopulation of cells that express the nerve growth factor (NGF) receptor trkA and the neuropeptide calcitonin gene-related peptide (CGRP). NGF increased BDNF within the trkA/CGRP cells to the extent that almost 90% of trkA cells contained BDNF mRNA after intrathecal NGF treatment, and 80-90% of BDNF-expressing cells contained trkA. Non-trkA cells that expressed BDNF included some trkC cells and some small cells that labeled with the lectin Griffonia simplicifolia IB4, a marker for cells that do not express trks. However, very few trkB cells expressed either BDNF mRNA or protein, and NGF did not increase BDNF expression in non-trkA cells. BDNF protein was anterogradely transported both peripherally and centrally. The central transport resulted in BDNF immunoreactivity in CGRP containing terminal arbors in the dorsal horn of the spinal cord, and this immunoreactivity was increased by NGF treatment. Electron microscopic analysis revealed that the BDNF immunoreactivity was present in finely myelinated and unmyelinated axons and in axon terminals, where it was most concentrated over dense-cored vesicles. Our data do not support an autocrine or paracrine role for BDNF within normal dorsal root ganglia, but indicate that BDNF may act as an anterograde trophic messenger. NGF levels in the periphery could influence dorsal horn neurons via release of BDNF from primary afferents.

Afferent Pathways↗

PubMed1997

TrkA immunoreactive neurones in the rat spinal cord.

We report the presence in rat spinal cord of a novel neuronal system expressing tyrosine kinase receptor (trkA), the high affinity receptor for nerve growth factor (NGF). TrkA immunoreactive cell bodies were observed in the intermediate grey matter of the spinal cord and were classified into three main groups: central canal cells located dorsolateral to the aqueduct, partition cells located between lamina X, and the lateral border of the intermediate grey, and a morphologically heterogeneous group which included large cells located near the lateral border. In situ hybridization confirmed that cells in all these areas express trkA mRNA. Combined immunofluorescence and retrograde Fluoro-Gold labelling was used to further characterise the projections and neurotransmitter profile of the trkA cells. Although often located in the vicinity of preganglionic cell groups, trkA immunoreactive cells are not themselves preganglionic. Rather, the central canal and partition cells belong to a neurochemically complex cholinergic propriospinal system. Many partition cells coexpress trkA, choline acetyltransferase (ChAT), the low affinity neurotrophin receptor, p75, and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d). In contrast, trkA immunoreactive central canal cells express ChAT, but do not express p75 and only a subpopulation express NADPH-d. The large trkA immunoreactive cells located on the lateral border do not express ChAT. TrkA immunoreactive fibres were also present and were located in the dorsal horn, in the dorsal columns, and in a bundle ventral to the aqueduct. However, double labelling revealed that the trkA immunoreactive fibres are not intrinsic but are primary afferent in origin and coexpress p75. The location of this novel trkA neuronal system is consistent with it having a role in the segmental integration of autonomic outflow. NGF could affect this system by modulating neuronal phenotype and/or synaptic efficacy.

Animals↗

PubMed1997

Constitutive expression of calmodulin-binding phosphoprotein GAP-43 in rat serotonergic and noradrenergic cell groups which project to the spinal cord.

In situ hybridization was combined with serotonin (5-hydroxytryptamine, 5-HT) or tyrosine hydroxylase immunocytochemistry and with Fluoro-Gold retrograde labeling of bulbo-spinal pathways in order to investigate the expression of GAP-43 mRNA in monoamine cell groups of the adult rat brain stem. Consistent with previous reports, GAP-43 mRNA was observed in serotonin and dopamine cell groups in the pons. In addition, GAP-43 expressing cells were observed in all the major monoamine cell groups in the medulla. Thus the B1, B2 and B3 serotonin cell groups all showed high GAP-43 expression in all contained many GAP-43 expressing serotonin cells with spinal cord projections. The A1, A2, A5 and A6 noradrenaline cell groups also showed high GAP-43 expression, although cells with spinal cord projections were largely restricted to the A5 group and A6 subcoeruleus region. In all areas, GAP-43 expressing cells with spinal cord projections were also observed which were not serotonergic or noradrenergic.

Animals↗

PubMed1997

Extraction of electrical characteristics from pixels of multifrequency EIT images.

Computer modelling has shown that electrical characteristics of individual pixels may be extracted from within multiple-frequency electrical impedance tomography (MFEIT) images formed using a reference data set obtained from a purely resistive, homogeneous medium. In some applications it is desirable to extract the electrical characteristics of individual pixels from images where a purely resistive, homogeneous reference data set is not available. One such application of the technique of MFEIT is to allow the acquisition of in vivo images using reference data sets obtained from a non-homogeneous medium with a reactive component. However, the reactive component of the reference data set introduces difficulties with the extraction of the true electrical characteristics from the image pixels. This study was a preliminary investigation of a technique to extract electrical parameters from multifrequency images when the reference data set has a reactive component. Unlike the situation in which a homogeneous, resistive data set is available, it is not possible to obtain the impedance and phase information directly from the image pixel values of the MFEIT images data set, as the phase of the reactive reference is not known. The method reported here to extract the electrical characteristics (the Cole-Cole plot) initially assumes that this phase angle is zero. With this assumption, an impedance spectrum can be directly extracted from the image set. To obtain the true Cole-Cole plot a correction must be applied to account for the inherent rotation of the extracted impedance spectrum about the origin, which is a result of the assumption. This work shows that the angle of rotation associated with the reactive component of the reference data set may be determined using a priori knowledge of the distribution of frequencies of the Cole-Cole plot. Using this angle of rotation, the true Cole-Cole plot can be obtained from the impedance spectrum extracted from the MFEIT image data set. The method was investigated using simulated data, both with and without noise, and also for image data obtained in vitro. The in vitro studies involved 32 logarithmically spaced frequencies from 4 kHz up to 1 MHz and demonstrated that differences between the true characteristics and those of the impedance spectrum were reduced significantly after application of the correction technique. The differences between the extracted parameters and the true values prior to correction were in the range from 16% to 70%. Following application of the correction technique the differences were reduced to less than 5%. The parameters obtained from the Cole-Cole plot may be useful as a characterization of the nature and health of the imaged tissues.

Biometry↗

PubMed1997

A Monte Carlo model for bone mineral measurement using dual energy X-ray absorptiometry.

A detailed Monte Carlo model designed to simulate bone mineral measurements using a typical K-edge DEXA technique (LUNAR DPX-L bone densitometer) has been developed. Factors considered in the model include measurement geometry, K-edge filtration of the X-ray spectrum, photon statistics, count rate corrections and detector resolution. Two semi infinite elliptical phantoms simulating the lumbar spine and the femoral neck respectively have been used. Each phantom consists of four tissue components-fat, lean soft tissue, cortical bone and trabecular bone. Model results display similar levels of accuracy and precision to those observed in clinical practice. The Monte Carlo model of bone mineral measurements provides a valid tool for study of the LUNAR DPX-L DEXA technique.

Absorptiometry, Photon↗

PubMed1996

Expression of trkA and p75 nerve growth factor receptors in the adrenal gland.

High and low affinity nerve growth factor (NGF) receptors were localized to specific cells and processes within the adult rat adrenal gland. Protein immunoreactivity and mRNA for high affinity trkA receptors were found in a subpopulation of adrenergic chromaffin cells and the clustered ganglion cells which do not express nitric oxide synthase (NOS). These ganglion cells, a subpopulation of the NOS containing ganglion cells, and all chromaffin cells were immunoreactive for p75, the low affinity neurotrophin receptor. TrkA immunoreactivity was found in ganglion cell processes and in sensory innervation but not preganglionic sympathetic innervation. The overlapping but distinct patterns of trkA and p75 expression provide a basis for differential actions of neurotrophins within the adrenal gland.

Adrenal Glands↗

PubMed1995

Expression of GAP-43 mRNA in preganglionic sympathetic neurones of the adult rat spinal cord.

In situ hybridization combined with intraperitoneal injection of the tracer Fluoro-Gold was used to study the distribution of GAP-43 mRNA in autonomic cell groups of the rat spinal cord. In the thoracic cord, numerous cells expressing GAP-43 mRNA were present in the central autonomic nucleus (lamina X) and in the intermediolateral cell column (IML). In the IML, all Fluoro-Gold-labelled preganglionic neurones were heavily labelled for GAP-43 mRNA. In contrast, Fluoro-Gold-labelled motoneurones were unlabelled or only lightly labelled. Our results indicate that preganglionic sympathetic neurones express GAP-43, a conclusion that is consistent with studies showing that GAP-43 immunoreactive terminals are present in autonomic ganglia. GAP-43 in these neurones may play a role in their continuing anatomical remodelling.

Animals↗

PubMed1995

Determination of equivalent x-ray spectra from attenuation data.

A method is described whereby a polyenergetic x-ray spectrum, as produced by a typical x-ray tube used in radiographic imaging, is approximated by an equivalent spectrum consisting of a small number of discrete x-ray photon energies, i.e. a line spectrum. The equivalent spectrum is found by a least squares fit to attenuation data for three different attenuating materials. Computer simulation and experimental measurements show that a three-energy equivalent spectrum accurately predicts the attenuation properties of the polyenergetic spectrum. The accuracy is limited by the uncertainty in the known values of the x-ray linear attenuation coefficients.

Computer Simulation↗

PubMed1993

Alpha 2-chimerin, an SH2-containing GTPase-activating protein for the ras-related protein p21rac derived by alternate splicing of the human n-chimerin gene, is selectively expressed in brain regions and testes.

n-Chimerin (alpha 1-chimerin) is a brain GTPase-activating protein (GAP) for the ras-related p21rac. We now report the occurrence of another form of chimerin, termed alpha 2-chimerin. This is the product of an alternately spliced transcript of the human n-chimerin gene encoding an N-terminal SH2 (src homology 2) domain in addition to the phorbol ester receptor and GAP domains. alpha 1- and alpha 2-chimerin mRNAs were expressed differently. In the rat brain, only alpha 1-chimerin mRNA was expressed in cerebellar Purkinje cells, although both alpha 1- and alpha 2-chimerin mRNAs occurred in neurons in the cerebral cortex, hippocampus, and thalamus. Only alpha 2-chimerin RNA was expressed in rat testes, in early pachytene spermatocytes. A 45-kDa SH2-containing chimerin corresponding to the alpha 2 form was purified from rat brain. As with Escherichia coli 45-kDa recombinant alpha 2-chimerin, purified brain alpha 2-chimerin exhibited racGAP activity which was stimulated by phosphatidylserine. The recombinant SH2 domain bound several 32P-labelled phosphoproteins of PC12 cells, whose phosphorylation increased in response to trophic factors, including nerve growth factor. To examine the relationships of alpha 1- and alpha 2-chimerin transcripts, human genomic DNA clones were characterized. In alpha 2-chimerin mRNA, a 3' splice acceptor site within exon 1 of alpha 1-chimerin mRNA was used, replacing its 5' untranslated region and N-terminal coding sequence. The single human n-chimerin gene was mapped to chromosome 2q31-q32.1, colocalizing with the CRE-BP1 transcription factor gene (2q32). It contained several splice junctions conserved with the sequence-related protein kinase C and bcr genes. alpha 2-Chimerin is only the second SH2-containing GAP and the first example of an SH2 domain generated by alternate splicing.

Alternative Splicing↗

PubMed1992

Developmental regulation and neuronal expression of the mRNA of rat n-chimaerin, a p21rac GAP:cDNA sequence.

Human n-chimaerin is a GTPase-activating protein (GAP) for p21rac and a phorbol ester receptor. We have isolated rat n-chimaerin cDNA and investigated the cellular and developmental pattern of mRNA expression in the brain. There is extensive sequence conservation with human n-chimaerin in the coding region and the first 400 nucleotides of the 3'-untranslated region (UTR) (90% and 83% identity respectively). The rat cDNA encodes an additional 35 N-terminal amino acids compared with the reported human cDNA, which has a 5'-UTR sequence inversion and a 41-nucleotide deletion including the putative initiator methionine. The rat cDNA encodes a 334-amino acid protein (38200 M(r), pI 8.04) with 97% amino acid sequence identity with the human protein, after correction of the human 5'-DNA sequence. n-Chimaerin mRNA was detectable in embryonic rat brain at day 15 and increased in amount postnatally from birth to 20 days, coincident with cellular differentiation and synaptogenesis. n-Chimaerin mRNA is restricted to neurons, with highest concentrations in hippocampal pyramidal cells, granule cells of the dentate gyrus and cortical neurons. In the cerebellum the mRNA was detected only in Purkinje neurons. The pattern and specificity of mRNA expression suggests an important role for n-chimaerin in neuronal signal-transduction mechanisms.

Amino Acid Sequence↗

PubMed1992

Tissue analysis using dual energy CT.

A new dual-energy x-ray CT algorithm is presented which makes use of both pre- and post-reconstruction data in an iterative manner to achieve accurate beam hardening correction and decomposition into basis materials. The technique does not require a calibration phantom and also does not require accurate estimation of the effective energies of the polyenergetic x-ray beams. It does however, require a knowledge of the incident x-ray spectra. For the situation where the incident spectra are unknown, a method is given whereby sufficiently accurate approximations to the spectra can be determined from attenuation measurements. Decomposition into basis materials makes use of spatial and energy information and an 'a priori' knowledge of the composition and attenuating properties of various tissue types. Any total number of basis materials may be used within the limitation that a subset containing a maximum of three materials is used for any individual pixel. Results of a computer simulation show that the algorithm produces accurate measurements of the concentrations of both high and low atomic number materials such as bone mineral, collagen, fat and air. However the results of a phantom study show that the measurement of the concentrations of low atomic number basis materials may be subject to systematic errors arising from uncertainties in the published values of linear attenuation coefficients.

Adipose Tissue↗

PubMed1992

Tissue analysis using dual energy CT.

Adipose Tissue↗