Biomedical subjects

M B Djamgoz

Publications and source records attributed to M B Djamgoz.

At least 37 records · Page 2Linked to original sources

PubMed1998

Expression and localization in the fish retina of a homologue of the Alzheimer's related PS1 gene.

Early-onset familial Alzheimer's disease (early-onset FAD) has been linked with mutations in the presenilin gene, PS1. Mutations in PS1 may affect the processing/ trafficking of beta-amyloid precursor-protein (betaAPP) and favour the production of toxic amyloid-beta fragments that are associated with neural degeneration. This study reports the expression of a PS1-like cDNA in the carp (Cyprinus carpio) retina (the encoded protein shows 76% identity to the human PS1 protein). Carp PS1 mRNA was localized by in situ hybridization to the photoreceptor cell, inner nuclear and ganglion cell layers. Expression of the PS1 gene in the rat retina was also confirmed. The retinal expression of PS1 raises the possibility that PS1 mutations also lead to neural degeneration in the retina.

Amino Acid Sequence↗

PubMed1998

Expression and localization in the fish retina of a homologue of the Alzheimer's related PS1 gene.

Early-onset familial Alzheimer's disease (early-onset FAD) has been linked with mutations in the presenilin gene, PS1. Mutations in PS1 may affect the processing/trafficking of b-amyloid precursor-protein (bAPP) and favour the production of toxic amyloid-b fragments that are associated with neural degeneration. This study reports the expression of a PS1-like cDNA in the carp (Cyprinus carpio) retina (the encoded protein shows 76% identity to the human PS1 protein). Carp PS1 mRNA was localized by in situ hybridization to the photoreceptor cell, inner nuclear and ganglion cell layers. Expression of the PS1 gene in the rat retina was also confirmed. The retinal expression of PS1 raises the possibility that PS1 mutations also lead to neural degeneration in the retina.

Alzheimer Disease↗

PubMed1998

Expression of skeletal muscle-type voltage-gated Na+ channel in rat and human prostate cancer cell lines.

Previous electrophysiological work has demonstrated expression of a voltage-gated Na+ channel (VGSC) specifically in two highly metastatic prostatic epithelial tumour cell lines: MAT-LyLu (rat) and PC-3 (human). However, the identity of the channel(s) present was uncertain. The present study used a combination of molecular biological techniques to demonstrate that full-length skeletal muscle type 1 (SkM1) VGSC mRNA is present in the mRNA pool of the MAT-LyLu cell line. mRNA for this particular channel type was also expressed in the PC-3 cells. In situ hybridisation data suggested that the level and pattern of rSkM1 mRNA expression were different in the Dunning cells of markedly different metastatic potential. Interestingly, the same type of mRNA was also detected in the weakly metastatic counterparts of the cells: AT-2 (rat) and LNCaP (human).

Animals↗

PubMed1998

Sodium channel protein expression enhances the invasiveness of rat and human prostate cancer cells.

Expression of Na+ channel protein was analysed in established cell lines of rat and human prostatic carcinoma origin by flow cytometry using a fluorescein-labelled polyclonal antibody. In many cell lines examined, the obtained frequency distribution profiles were bimodal and identified a subpopulation of cells which expressed high levels of Na+ channel protein. A significant positive correlation was demonstrated between the proportion of channel-expressing cells and the functional ability of individual cell lines to invade a basement membrane matrix in vitro. In addition, two transfectant cell lines containing rat prostate cancer genomic DNA were found to express significantly elevated levels of Na+ channel protein when compared with the original benign recipient cell line. Enhanced Na+ channel expression by two metastatic derivatives of these transfectant cells directly correlated with increased invasiveness in vitro. These studies strongly support the hypothesis that expression of Na+ channel protein and the metastatic behaviour of prostatic carcinoma cells are functionally related, either by endowing the membranes of these cells with specialised electrophysiological properties (e.g. enhancing their motility and/or secretory activities) and/or by perturbing endogenous mechanisms regulating ionic homeostasis within the cells.

Animals↗

PubMed1998

Electrophysiological characterization of voltage-gated Na+ current expressed in the highly metastatic Mat-LyLu cell line of rat prostate cancer.

Voltage-gated Na+ channels, classically associated with impulse conduction in excitable tissues, are also found in a variety of epithelial cell types where their possible functions are not known so well. We have previously reported expression of a voltage-gated Na+ channel specifically in the highly metastatic Mat-LyLu rat prostate cancer cell line; blockage of the current with tetrodotoxin (TTX) significantly reduced the invasiveness of the cells in vitro, suggesting that the channel may have a functional role in metastasis. The aim of the present study was to characterize this current using the whole-cell patch clamp recording technique, and compare it to Na+ currents found in various other tissues. The inward current of the Mat-LyLu cells was abolished completely, but reversibly, in Na+-free solution, confirming that Na+ was indeed the permeant ion. Activation occurred at -40 mV and currents reached a maximal amplitude at around 6 mV. Boltzmann fits to current activation and steady-state inactivation revealed that the currents were half activated at about -15 mV and half inactivated at -80 mV. Both current inactivation and recovery from inactivation followed a double-exponential time course with fast and slow components. The Na+ currents were highly sensitive to block by TTX (IC50 approximately 18 nM), whilst 1 microM mu-conotoxin GIIIA mostly had no effect. 100 microM Cd2+ also had no effect on the current, whilst 2.5 mM Cd2+, Mn2+, and Co2+ each caused a depolarizing shift in activation and a reduction in peak conductance of around 20%. In conclusion, the Na+ channel expressed in the highly metastatic Mat-LyLu cell line appeared to have electrophysiological and pharmacological properties of TTX-sensitive channels. Further work is needed, however, to elucidate the exact nature of the channel protein and the mechanism(s) of its involvement in cellular invasiveness.

Animals↗

PubMed1998

Modulation of chromatic difference in receptive field size of H1 horizontal cells in carp retina: dopamine- and APB-sensitive mechanisms.

Chromatic aspects of receptive field size in the H1 horizontal cell syncytium of the carp retina were investigated using spectral photostimuli (blue or red) presented in the form of either a pair of a small spot and annulus, or a narrow moving slit. In the light-adapted retina, the receptive field for the blue stimulus was found to be significantly smaller than that for the red, i.e. there was a chromatic difference in the receptive field size. During the course of dark adaptation, the overall receptive field size increased, but the chromatic difference decreased. Immediately after adaptation to bright light, the receptive field sizes were reduced significantly, but the chromatic difference increased, mainly due to a greater reduction in the receptive field for the blue stimulus. Application of dopamine (5 microM) to a dark-adapted retina gradually decreased the receptive field size for both colours, but the chromatic difference became larger, again due to a greater reduction in the receptive field size for the blue stimulus. 2-Amino-4-phosphonobutyrate (APB) applied to light-adapted retinae at a working concentration of 1 mM, greatly expanded the receptive field size and suppressed the chromatic difference due to the effect being greater for the receptive field for the blue stimulus. The effect of APB was slow and cumulative. On the other hand, intracellular injection of cGMP or dibutyryl-cGMP increased the chromatic difference in the receptive field size. It is suggested (i) that the chromatic difference in the receptive field size could be due to a cGMP-coupled, conductance-decreasing receptor mechanism activated by APB; and (ii) that the mechanism is associated with short-wavelength sensitive cone input to the H1 cells and operates in the light-adapted state of the retina.

Adaptation, Ocular↗

PubMed1998

Cloning and functional characterisation of two novel nicotinic acetylcholine receptor alpha subunits from the insect pest Myzus persicae.

Nicotinic acetylcholine receptors play a major role in excitatory neurotransmission in insect CNSs and constitute an important target for insecticides. Here, we report the isolation and functional characterisation of two cDNAs encoding nicotinic acetylcholine receptor alpha subunits from a major insect pest, the peach-potato aphid Myzus persicae. These two subunits, termed Mp alpha1 and Mp alpha2, are respective structural homologues of the Drosophila D alpha2/Schistocerca gregaria alphaL1 alpha-subunit pair and the Drosophila ALS alpha subunit. Xenopus oocyte expression confirmed that each Myzus subunit can form functional acetylcholine- or nicotine-gated channels. However, some electrophysiological and pharmacological properties of the Myzus subunits were distinct from those encoded by the corresponding Drosophila subunits. Coexpression of the Myzus subunits with the chick beta2 subunit revealed other differences from the Drosophila system, as only very limited potentiation of agonist-induced currents was observed with Mp alpha2 and none with Mp alpha1. Available data therefore indicate that structurally homologous insect nicotinic acetylcholine receptor alpha subunits from different species can exhibit distinctive physiological and pharmacological characteristics.

Amino Acid Sequence↗

PubMed1998

Dopamine receptor subtypes expressed in vertebrate (carp and eel) retinae: cloning, sequencing and comparison of five D1-like and three D2-like receptors.

Eight dopamine receptor-like cDNA clones were isolated from the carp (Cyprinus carpio) retina and four dopamine receptor-like cDNA clones were isolated from the European eel (Anguilla anguilla) retina. These cDNA clones show high sequence and structural homology to the known dopamine receptor subtypes. The sequence similarity and phylogenetic analysis revealed that five subtypes (D1A3, D1A4, D1B, D1C and D1X) in the carp retina and four subtypes (D1A1, D1A2, D1B and D1C) in the eel retina are D1-like receptor subtypes, and three (D2, D4A and D4B) in the carp retina are D2-like receptor subtypes; no D2-like receptor was found in the eel. Carp D1A3 and D1A4, carp D4A and D4B, and eel D1A1 and D1A2 are highly homologous pairs of receptors which show significant, domain-specific differences to each other and to their species homologues. The structure of the third cytoplasmic loop in the carp D1X receptor was particularly different from the other D1-like receptors. The implications of these structural differences in terms of dopamine receptor activation and signalling are discussed. It is suggested that the known diverse physiological and pharmacological effects of dopamine on the retinal neurones are likely to be mediated through these multiple receptor subtypes which may be coupled to different signal transduction pathways.

Amino Acid Sequence↗

PubMed1997

Effects of ageing on spatial aspects of the pattern electroretinogram in male and female quail.

This study deals with age-related functional changes in the eye of quail. Measurements of lens refractive state and transmission, as well as pupil diameter, showed that the quail's eye became more myopic with increasing age, but no change could be detected in lens transmission and pupil diameter. Three aspects of the pattern electroretinogram (PERG) were studied after correcting for ocular refraction: (i) spatial characteristics (visual acuity and contrast sensitivity); (ii) maximal amplitude; and (iii) peak latency. The PERG results suggested that the visual acuity was age-independent for both sexes. However, the contrast sensitivity of the old quail (16-20 months) was much lower (1.5-3 times) than that of the young (3-6 months) at low to intermediate (< 2 c/d), but not at high spatial frequencies. The peak latency of the PERG response was 3-4 msec longer in the old birds compared with the young, while the maximal amplitude of the PERG response was age-independent. These results clearly suggested that at least some of the age-induced changes are located in the neural retina.

Aging↗

PubMed1997

Retinal dopamine depletion in young quail mimics some of the effects of ageing on visual function.

The hypothesis that retinal dopaminergic (DA) neurones are involved in the visual functions of interest was tested. The retinal DA in young quail was partially depleted by intravitreal injection of 6-hydroxydopamine (6-OHDA). It was found that the refractive state of 6-OHDA-treated birds became more myopic than normal (untreated) young, whereas the pupil diameter was not affected. The contrast sensitivity of 6-OHDA treated quail was significantly lowered (two to three times) at all spatial frequencies studied (0.25-5 c/d), and the peak latency of pattern electro-retinogram (PERG) response was prolonged by 3-4 msec (9%). Furthermore, the visual acuity and maximal amplitude of PERG response of the 6-OHDA-treated young quail were lower than those of normals. From histochemical studies, it was revealed that the morphology of the DA cells of 6-OHDA-treated young appeared similar to those of the old quail; the DA cells of 6-OHDA-treated retinae were less fluorescent and 2.5-5 times less numerous than respective controls. Combining the PERG and the morphological results, it would seem that the retinal DA plays an important role in the visual functions studied, and that loss of retinal DA could underlie some of the visual changes which occur during ageing.

Adrenergic Agents↗

PubMed1997

Neurobiology of retinal dopamine in relation to degenerative states of the tissue.

Neurobiology of retinal dopamine is reviewed and discussed in relation to degenerative states of the tissue. The Introduction deals with the basic physiological actions of dopamine on the different neurons in vertebrate retinae with an emphasis upon mammals. The intimate relationship between the dopamine and melatonin systems is also covered. Recent advances in the molecular biology of dopamine receptors is reviewed in some detail. As degenerative states of the retina, three examples are highlighted: Parkinson's disease; ageing; and retinal dystrophy (retinitis pigmentosa). As visual functions controlled, at least in part, by dopamine, absolute sensitivity, spatial contrast sensitivity, temporal (including flicker) sensitivity and colour vision are reviewed. Possible cellular and synaptic bases of the visual dysfunctions observed during retinal degenerations are discussed in relation to dopaminergic control. It is concluded that impairment of the dopamine system during retinal degenerations could give rise to many of the visual abnormalities observed. In particular, the involvement of dopamine in controlling the coupling of horizontal and amacrine cell lateral systems appears to be central to the visual defects seen.

Aging↗

PubMed1997

Ionic effects of the Alzheimer's disease beta-amyloid precursor protein and its metabolic fragments.

Alzheimer's disease is a progressive dementia characterized in part by deposition of proteinaceous plaques in various areas of the brain. The main plaque protein component is beta-amyloid, a metabolic product of the beta-amyloid precursor protein. Substantial evidence has implicated beta-amyloid (and other amyloidogenic fragments of the precursor protein) with the neurodegeneration observed in Alzheimer's disease. Recently, beta-amyloid precursor protein and its amyloidogenic metabolic fragments have been shown to alter cellular ionic activity, either through interaction with existing channels or by de novo channel formation. Such alteration in ionic homeostasis has also been linked with cellular toxicity and might provide a molecular mechanism underlying the neurodegeneration seen in Alzheimer's disease.

Alzheimer Disease↗

PubMed1997

Nitric oxide, 2-amino-4-phosphonobutyric acid and light/dark adaptation modulate short-wavelength-sensitive synaptic transmission to retinal horizontal cells.

Light-induced changes in the input resistance (Rin) of external, luminosity (i.e. H1) type horizontal cell (HC) perikarya were studied by the bridge-balance method in light-adapted and dark-adapted retinae of carp. Changes in input resistance (delta Rin) induced by short-(460 nm) and long-wavelength (674 nm) flashes, adjusted in intensity to elicit equal-amplitude membrane voltage responses (equal-voltage condition), were measured. In light-adapted retinae, long-wavelength stimuli increased Rin consistently; in contrast, the increase was much less with short-wavelength stimuli. This equal-voltage chromatic delta Rin difference was lost in dark-adapted retinae whereby the delta Rin (an increase) became the same for short- and long-wavelengths. The chromatic delta Rin difference could be recovered by light adaptation or application of sodium nitroprusside to the dark-adapted retinae. Conversely, the equal-voltage chromatic delta Rin difference was eliminated by injection of NG-monomethyl-L-arginine into H1HCs of the light-adapted retinae or by treating the retinae with 2-amino-4-phosphonobutyrate (APB). These results suggest that H1HCs of the carp retina possess distinct postsynaptic mechanisms which mediate short- and long-wavelength signal transmission. Furthermore, it appears that the short-wavelength-sensitive pathway is active only during the light-adapted state of the retina. Taken together, therefore, the short-wavelength transmission to H1HCs probably operates on an APB-sensitive glutamate receptor, with nitric oxide as a light-adaptive messenger.

Acclimatization↗

PubMed1997

G-protein activation, intracellular Ca2+ mobilization and phosphorylation studies of membrane currents induced by AlF4- in Xenopus oocytes.

We have examined the electrophysiological responses induced by aluminium fluoride (AlF4-) and carbachol in Xenopus oocytes. Application of AlF4- induced Ca(2+)-dependent oscillatory and smooth Cl- currents. Pre-treatment of oocytes with microinjected guanosine 5'-O-(2-thiodiphosphate) diminished the currents, indicating that the effect of AlF4- occurred through G-protein activation. Confocal imaging of intracellular Ca2+ clearly demonstrated that AlF4- could increase the internal Ca2+ concentration in oocytes in the absence of external Ca2+. A protein kinase (PK) activator (4-beta-phorbol 12,13-dibutyrate) decreased the AlF4(-)-induced membrane currents, whereas a PK inhibitor (staurosporine) caused an increase. On the other hand, the protein phosphatase inhibitor (okadaic acid) showed little effect. Although the effects of the phosphorylating/dephosphorylating agents on the carbachol-induced currents were qualitatively similar to the case of AlF4-, some quantitative differences was noted. The results are discussed in terms of the signaling pathways involving muscarinic receptors and G-protein(s) in Xenopus oocytes.

Aluminum Compounds↗

PubMed1997

Expression and functional analysis of voltage-activated Na+ channels in human prostate cancer cell lines and their contribution to invasion in vitro.

Ion channels are important for many cellular functions and disease states including cystic fibrosis and multidrug resistance. Previous work in the Dunning rat model of prostate cancer has suggested a relationship between voltage-activated Na+ channels (VASCs) and the invasive phenotype in vitro. The objectives of this study were to 1) evaluate the expression of VASCs in the LNCaP and PC-3 human prostate cancer cell lines by Western blotting, flow cytometry, and whole-cell patch clamping, 2) determine their role in invasion in vitro using modified Boyden chambers with and without a specific blocker of VASCs (tetrodotoxin). A 260-kd protein representing VASCs was found only in the PC-3 cell line, and these were shown to be membrane expressed on flow cytometry. Patch clamping studies indicated that functional VASCs were present in 10% of PC-3 cells and blocking these by tetrodotoxin (600 nmol/L) reduced their invasiveness by 31% (P = 0.02) without affecting the invasiveness of the LNCaP cells. These results indicate that the reduction of invasion is a direct result of VASC blockade and not a nonspecific action of the drug. This is the first report of VASCs in a human prostatic cell line. VASCs are present in PC-3 but not LNCaP cells as determined by both protein and functional studies. Tetrodotoxin reduced the invasiveness of PC-3 but not LNCaP cells, and these data suggest that ion channels may play an important functional role in tumor invasion.

Animals↗

PubMed1996

Voltage dependency of light-evoked on-off transient amacrine cell responses in carp retina.

The voltage dependency of the ON and the OFF components of transient amacrine cell responses was studied using two-electrode voltage-clamp and current-clamp techniques in the isolated retina of the carp. The two independent approaches gave similar data. When cells were voltage clamped near their resting potentials, both response components were associated with transient inward currents. Hyperpolarization increased response size (current or voltage) whilst depolarization decreased it. Response reversal, or a tendency for it, occurred at membrane potentials significantly more positive than the resting level with some quantitative variability. These data support the view that the ON-OFF depolarizations represent basically excitatory postsynaptic potentials and that the transience of the responses cannot mainly be due to any voltage-dependent conductance.

Animals↗

PubMed1996

Molecular physiology, biochemistry, and pharmacology of Alzheimer's amyloid precursor protein (APP).

The function of APP is not yet known in detail but growing evidence exists that APP may mediate cell interactions with the cell surface or soluble glycoproteins and defense mechanisms in the CNS involving the immune system. We describe here the finding that almost all CD4+ lymphocytes and the majority of CD8+ lymphocytes were positive for A beta and the antibodies against A beta or APP did not inhibit the [3H]-thymidine uptake of mitogen-treated lymphocytes significantly. There were no differences in the A beta immunoreactivity on the cell surface of lymphocytes between Alzheimer's disease (AD) and control samples. Excessive amyloidogenic pathway of APP processing may be the final common pathway involved in the pathogenesis of AD. Thus, the identification of proteases or factors leading to aberrant proteolysis which process APP to yield a variety of potentially amyloidogenic fragments would promise pharmacological targets to develop anti-AD drugs. In attempts to define the proteases or factors which alter the balance between nonamyloidogenic and amyloidogenic processing pathways, our study indicates that thrombin or acetylcholinesterase(AChE)-associated protease may be involved in the amyloidogenic processing pathway of APP in vivo to generate amyloidogenic intermediates linked to amyloid deposition. Highly specific and dose-dependent direct modulation of APP processing by biologically available metal ions including Ca2+, Zn2+, Fe2+/Fe3+ and Al3+ suggest the disrupted metal homeostasis as factors leading to overaccumulation of APP and subsequent aberrant proteolysis utilizing excessive amyloidogenic processing pathway. There is mounting evidence that at least some of the neurotoxicity associated with AD is due to fragments from APP. Most research has focused on the toxic effect and the ion channel activity of A beta in causation of the disease. The possible role of other cleaved products of APP is less clear. We investigated the channel-forming ability of various products of APP when applied to Xenopus oocytes and their neurotoxicity in vitro. CT105 peptide was found to be exceedingly potent at 500 nM concentration in forming nonselective ion channels during application from either outside or inside the oocyte and more toxic than either of the A beta fragments, A beta 25-35, or A beta 1-40. Taken together, these results suggest the possible involvement of CT peptide in inducing the neurotoxicity characteristic of AD through the direct damage on the cell membrane. Therefore, we hypothesize that amyloidogenic CT may make nonselective ion channels or pores in the membrane and may cause neuronal death in the early stage of AD and then further metabolized to more stable and less toxic A beta which may be finally deposited in the brain where it could inflict further toxicity to neurons. Here we report successful inhibition of APP gene expression by antisense oligodeoxynucleotides at the mRNA or the protein level in in vitro and cell culture systems.

Alzheimer Disease↗

PubMed1996

Effects of different patterns of light adaptation on cellular and synaptic plasticity in teleost retina: comparison of flickering and steady lights.

The importance of the pattern of light stimulus in inducing light-adaptive morphological (cellular and sub-cellular) changes in the outer retina of the carp was assessed. Thus, the effects of steady and flickering backgrounds (of the same intensity) on cone photomechanical movements (PMMs) and horizontal cell (HC) spinules were compared by quantitative measurements. For both phenomena, flickering stimuli had a significantly greater effect than steady, although the number of photons delivered by the former was one-half. The results demonstrate clearly that the pattern of light stimulus is indeed an important parameter in determining the extent of adaptation in the outer retina. The effects are discussed in relation to possible neurochemical bases of light-adaptive neural control mechanisms.

Adaptation, Physiological↗