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Biomedical subjects

P L Marchiafava

Publications and source records attributed to P L Marchiafava.

At least 19 recordsLinked to original sources
PubMed2002

Retinal photoreceptors of Syrian hamsters undergo oxidative stress during streptozotocin-induced diabetes.

AIMS/HYPOTHESIS: The aim of this study was to verify whether retinal photoreceptors, like other tissues, are subject to oxidative stress during diabetes. METHODS: Oxidative stress was monitored by the oxidation of preloaded dehydrorhodamine 123 into fluorescent rhodamine 123, during a period of intense illumination of isolated rod retinal receptor cells. These were obtained from 22 Syrian hamsters injected with streptozotocin (50 mg/kg body weight., intraperitoneal route) 90 days before the study began. Eleven hamsters were treated daily with melatonin (0.4 mg/kg body wt., per os), an antioxidant synthesized within photoreceptors. Isolated photoreceptors were bathed on the stage of a Leitz Orthoplan microscope, where the fluorescent lamp also served as the light stimulus (485 nm). Fluorescence irradiance was measured by photometry and stored in a personal computer for further analysis. RESULTS: The light-induced oxidant production greatly decreased and was also delayed in the streptozotocin-injected hamsters compared with the control hamsters matched for age. Similar effects were obtained in control photoreceptors after 40 min incubation with 2-2'-azobis (2-amidinopropane) dihydrochloride, a potent lipoperoxidation inducer. The effect of melatonin was to partially restore the light-induced fluorescence response. CONCLUSION/INTERPRETATION: The depression of the light-induced oxidative response in diabetic photoreceptors could be ascribed to a hyperglycaemia-induced background of oxidative stress whereby the light-oxidizable substrate is actually lowered. Melatonin induces a larger fluorescence response during illumination, probably as a consequence of its antioxidant effect during diabetes, which would provide more oxidizable lipids.

Amidines↗
PubMed2001

Phentolamine suppresses the increase in arteriolar vasomotion frequency due to systemic hypoxia in hamster skeletal muscle microcirculation.

Systemic hypoxia (8%, 11% and 15% oxygen gas mixture inspiration) has been shown to increase the frequency of arteriolar rhythmic diameter changes in hamster skeletal muscle microcirculation. The effects of phentolamine on vasomotion frequency during systemic hypoxia were studied in Syrian hamsters implanted with a plastic chamber in the dorsum skin. Phentolamine (50 microg/100 g body wt.) was injected intravenously before the 20-min exposure to 11% oxygen gas mixture. The microvessels were studied with a fluorescent microscopy technique, using fluorescein isothiocyanate bound to dextran (mol. wt. 150,000) as a tracer. Vessel diameters were measured with a shearing method. Fourier transform and autoregressive modeling were used to assess the time variant features of diameter changes. Under baseline conditions, the arterioles were characterized by rhythmic diameter changes with fundamental frequency related to vessel size. The terminal branchings were dominated by order 3 vessel activity (frequency: 0.08-0.16 Hz) spreading downstream to all daughter arterioles. Systemic hypoxia caused an increase in vasomotion frequency of order 3 arterioles up to 0.3-0.5 Hz (average: 0.40 +/- 0.06 Hz) and a significant decrease in mean diameter (-28 +/- 5%). Phentolamine completely suppressed the rhythmic changes in diameter of order 3 arterioles that dilated significantly (+ 30 +/- 4%). Therefore, the effects of systemic hypoxia on arteriolar vasomotion appear to be triggered by an increase in sympathetic nervous discharge that induces a rise in frequency up to 0.3-0.5 Hz.

Animals↗
PubMed1999

Regulation of Bcl-2 protein expression during oxidative stress in neuronal and in endothelial cells.

The relationship between oxidative stress and Bcl-2 expression was investigated in two different experimental models of oxidative stress. Acute oxidative stress was assessed by measuring, with fluorescence microscopy and cytofluorimetry, the increase in fluorescence of the oxidation-sensitive probe dihydrorhodamine 123, both in retinal rod receptor cells exposed to bright light (0.32 mW/cm(2) for 15 minutes) and in human endothelial cells treated with the immunosuppressant cyclosporin A (200 microM for 21 h). In both cell types, acute oxidative stress reduced Bcl-2 expression and also caused a significant increase in the level of nucleosomes. Interestingly, chronic treatment with clinical concentrations of cyclosporin A (0.5-2.5 microM for 8 days) led to a significant increase in Bcl-2 expression, while nucleosomes were similar to control level. This suggests that up-regulation of Bcl-2 protein by low levels of oxidants may represent a critical factor in cellular adaptation to drug toxicity.

Endothelium, Vascular↗
PubMed1999

Melatonin as an antioxidant in retinal photoreceptors.

Dark-adapted, single photoreceptors isolated from the frog retina produce reactive oxygen species (ROS) after about 1 min of illumination with saturating light that we verified by their oxidation of preloaded dihydrorhodamine 123 (DHR) into the fluorescent rhodamine 123 (RHO). In this preparation we tested the antioxidant effects of vitamin E and of melatonin. Melatonin at picomolar and low nanomolar concentrations was determined to be 100 times more potent in inhibiting the light-induced oxidative processes than was vitamin E. On the contrary, both compounds exerted potent prooxidant effects at micromolar concentrations that is above the physiological levels of melatonin. This provides evidence that physiological concentrations of melatonin in a living cell may exert protective actions against a natural oxidant stimulus (light). This helps to define the functional role of endogenous melatonin in photoreceptors, which by their physiological characteristics, are among the marked producers of ROS in the organism.

Animals↗
PubMed1998

Effects of melatonin on lipid peroxidation induced by oxygen radicals.

We here report the activity of the neurohormone melatonin (MLT) as a scavenger of free radicals in two different experimental models: (a) linoleic acid peroxidation initiated by different free radical-generating systems and (b) a multilamellar vesicle system composed of dilinoleoylphosphatidylcholine. In system (a) linoleic acid peroxidation, induced by either the water-soluble initiator 2,2'-azobis (2-amidinopropane) dihydrochloride (ABAP) or Fe2+-EDTA addition to 2.6 mM linoleic acid dispersed in SDS-phosphate buffer, was evaluated as the formation of conjugated dienes, measured spectrophotometrically at 236 nm. MLT did not reduce the rate of peroxidation induced by ABAP, but did reduce, in a concentration-dependent fashion, the rate of the reaction activated by Fe2+-EDTA. In system (b) multilamellar vesicles were used as the substrate for lipid peroxidation, initiated by Fe2+-EDTA and determined by means of malonaldehyde (MDA) and 4-hydroxyalkenal (4-HDA) content. MLT was found to be slightly more effective in system (b) than in the dispersed linoleic acid system (see a). These results show that MLT inhibits lipid damage induced by oxygen free radicals. However, MLT is only about one one-hundredth as effective an antioxidant as vitamin E in the micelles system.

Amidines↗
PubMed1997

Melatonin induces membrane conductance changes in isolated retinal rod receptor cells.

Experiments were conducted to verify whether the neurohormone melatonin influences the membrane conductance of photoreceptors isolated from the frog retina. It has been found that 20 microM melatonin decreases membrane conductances both in the linear and non linear ranges by <0.4 nS. These actions are estimated to produce in dark adapted photoreceptors an increase of the response to a dim light induced change of the dark current of about 21%, i.e. from 1.3 to 1.62 mV/pA.

Animals↗
PubMed1997

Cell physiology of the pineal body.

The results from recent experiments on the cellular physiology of the trout pineal photoreceptors are briefly reviewed. The arguments are mainly concerned with pineal phototransduction. These studies have stimulated further research on melatonin, a molecule produced in pineal as well as in retinal photoreceptors. A discussion follows on our actual research object, that is a study of the influences of endogenous melatonin upon retinal receptor cells activities.

Animals↗
PubMed1996

Melatonin prevents ischemia reperfusion injury in hamster cheek pouch microcirculation.

OBJECTIVE: We used the hamster cheek pouch microcirculation to investigate the effects of melatonin (ME) on ischemia reperfusion (I-R) injury by in vivo microscopy. ME is a hormone produced by the pineal gland and is the most powerful and effective hydroxyl radical scavenger detected to date in vitro. The second aim was to determine the scavenger effect of ME in cheek pouch microcirculation when topically applying an oxygen-derived free radical generating system. METHODS: Ischemia was induced by clamping the cheek pouch for 30 min followed by 30 min of reperfusion. We quantified the increase in permeability, the perfused capillary length and leukocyte adhesion by computerized methods. Microcirculation was also exposed to a hypoxanthine-xanthine oxidase (H-X) system. RESULTS: In control hamsters I-R was associated with increased permeability, increased number of leukocytes sticking to venules, and decreased perfused capillary length. Treatment with ME completely inhibited microvascular edema formation and reduced the number of leukocytes sticking to venules after reperfusion. Moreover, ME prevented the marked decrease in perfused capillary length, preserving microvascular perfusion. ME topically applied reduced significantly the permeability increase due to H-X exposure. CONCLUSIONS: The beneficial effect of ME may be related to its antioxidant properties. These protect the endothelial barrier integrity as well as preserve microvascular blood perfusion by dysfunctions after I-R.

Animals↗
PubMed1993

The electrical responses of the trout pineal photoreceptors to brief and prolonged illumination.

Intracellular recordings from 103 photoreceptors in the excised pineal body of adult trouts were obtained by using single electrode current- and voltage-clamp techniques. The photoresponses to brief flashes showed the same polarity but a slower time course than those previously recorded from retinal photoreceptors of lower vertebrates. Pineal photoreceptors showed spectral sensitivity peaks at about 495 and 521 nm and absolute sensitivity comparable to retinal cone cells of the same species. The photoreceptor membrane conductance, measured under voltage clamp during moderate illumination was about 10% lower than in the dark, and the extrapolated reversal potential of the response was at 60 mV above the dark membrane potential. The addition of 3-isobutyl-1-methylxanthine (IBMX) to the perfusate was followed by a receptor depolarization in the dark and by a slow-down of the response kinetic. Pineal receptor cells produce constant amplitude responses during steady illumination, without displaying the delayed slow depolarization typically associated with light adaptation of retinal photoreceptors. Photoresponses to brief flashes superimposed on a steady illumination are decreased in amplitude by an amount directly related to the background intensity. Increase of the background intensity leads to threshold increments without significant changes of the saturation intensity, resulting in a gradual compression of the cell dynamic range. These results were discussed relative to light adaptation in retinal photoreceptors. The conclusion can be drawn that the response properties of pineal photoreceptors during steady illumination are part of an unknown, self-regulating mechanism to lock the rate of metabolism and secretion of indolamines to the absolute level of diurnal light.

1-Methyl-3-isobutylxanthine↗
PubMed1989

Membrane resistance properties of pineal photoreceptors in the trout.

Intracellular recordings were obtained from pineal photoreceptors of the trout Salmo irideus. Illumination produced graded hyperpolarizing responses whose time course resembled that of retinal cones in lower vertebrates. Current-voltage relations were derived by using a voltage-clamp technique in the dark and during illumination. Membrane resistance values obtained in the dark were about 550 M omega, and they increased during illumination. The extrapolated reversal potential of the photocurrent is about 55 mV above the dark membrane potential, indicating that an ionic mechanism similar to that in retinal photoreceptors may be involved.

Animals↗
PubMed1986

Subsurface cisternae in retinal double cones.

Double cones of tench and goldfish retina are characterized by extensive subsurface cisternae underlying the plasma membranes at the appositional area between the principal and accessory cone. Such a membrane system is absent in double cones of turtle and salamander retina. Measurements on both transverse and longitudinal sections gave a total appositional area of about 75 square microns, the subsurface cisterna in each element of the double cone being around 8-10% smaller due to multiple fenestrations at the level of the paraboloid. No gap junctions joining the inner segments of tench and goldfish double cones were detected, while gap junctions could be observed at the level of the ellipsoid and paraboloid of turtle double cones. The possible role of the subsurface cisternae in functional interactions between double cone elements is discussed.

Animals↗
PubMed1985

Intracellular recording from single and double cone cells isolated from the fish retina (Tinca tinca).

Intracellular recording were obtained from isolated single and double cone cells of the tench retina. Photoresponses show features characteristic of other species and behave linearly with very dim illumination. The cells spectral sensitivity matches their pigment absorption spectrum measured by previous authors. The principal and accessory members of double cones show a maximal sensitivity peak at about 644 and 547 nm, respectively. In addition, each of the two action spectra shows a secondary inflection at the peak wavelength of the adjacent member, suggesting functional coupling between the two members of double cones.

Animals↗
PubMed1983

An "antagonistic" surround facilitates central responses by retinal ganglion cells.

Ganglion cells in the turtle retina respond to increments of saturating illumination of the receptive field centre by progressively decreasing the number of spikes of the responses, perhaps as a result of membrane inactivation. Intracellular recording indicates that a simultaneous illumination of the receptive field surround greatly facilitates the "suprasaturated" central responses, while the expected centre-surround antagonism is still present between photoresponses below saturation. It is suggested that both pre- and post-synaptic mechanisms provide the ganglion cells with the unique possibility, among other retinal cells, to shift their full dynamic range across more than 3 log units of illumination intensity.

Action Potentials↗
PubMed1983

The organization of inputs establishes two functional and morphologically identifiable classes of ganglion cells in the retina of the turtle.

Analyses of the photoresponses and morphologies of ganglion cells in the turtle retina suggests that these cells are of two types: type A ganglion cells receive predominantly bipolar inputs, while type B ganglion cells receive a mixture of inputs from bipolar and amacrine cells. According to these classification criteria, ganglion cells may reflect either activities of the outer or of the inner plexiform layer, respectively. To corroborate the validity of this classification a number of basic properties of ganglion cells were analysed. It was found that double colour opponency was characteristic of type A cells and directional selectivity was performed only by type B cells. In addition, the absolute sensitivity and the firing rate of type A cells were significantly higher than those of type B cells.

Animals↗
PubMed1982

The photoresponses of structurally identified amacrine cells in the turtle retina.

Intracellular recordings were obtained from amacrine cells afterwards identified morphologically by horseradish peroxidase injection. There is a correlation between the time course of the photoresponses and the distribution of the cell processes across the inner plexiform layer (i.p.l.). Cells producing the shortest duration, transient 'on-off' photoresponses branched in a single, narrow stratum of the i.p.l. (3-7 microns across). Transient photoresponses with a longer time course were recorded from cells branching in a thicker stratum of i.p.l. (up to 20 microns), or from bistratified cells. Amacrine cells producing sustained centre-on or centre-off photoresponses were radially diffused across the whole i.p.l.; therefore this type of photoresponse need not be associated with a specific cellular stratification within the i.p.l. It is concluded that the two main functional types of amacrine cell, i.e. transient on-off and sustained centre-on and centre-off, are subject to different structural organization of inputs than are the homologous physiological types of ganglion cells in this species, in the cat and in the carp. In a summary diagram the observed characteristics of the photoresponses are tentatively explained in terms of a non-homogeneous distribution of bipolar synaptic inputs along amacrine cell processes.

Animals↗
PubMed1982

Electrophysiological and structural studies of the organization of inputs to ganglion cells in the turtle retina.

The structure of electrophysiologically identified bipolar, amacrine and ganglion cells has been studied in the turtle retina. The distribution of these cells processes within the inner plexiform layer (i.p.l.) was evident after intracellular injection of horseradish peroxidase (HRP) or Procion yellow. The observations indicated that ganglion cells may be subdivided into two groups, on the basis of their organization of inputs. Type A ganglion cells receive only bipolar cells inputs, and Type B ganglion cells receive an additional input from amacrine cells. Each of the two groups of ganglion cells performs specific visual functions, like color discrimination or directional sensitivity.

Animals↗
PubMed1981

Interactions leading to colour opponency in ganglion cells of the turtle retina.

In the turtle retina, colour-dependent photoresponses could be recorded intracellularly from ganglion cells receiving only bipolar cell input. Thus, the mechanism for colour discrimination by these ganglion cells (type A) is contained in the outer plexiform layer of the retina and depends on interaction between horizontal and cone cells. Ganglion cells receiving an additional amacrine input (type B) are not influenced by colour, and have about 0.7 logarithmic unit lower absolute sensitivity to peak wavelength than have type A ganglion cells.

Animals↗