Pial microvascular responses to transient bilateral common carotid artery occlusion: effects of hypertonic glycerol.

Objective: The aim of the study was to assess the rat pial microvessel alterations due to transient bilateral common carotid artery occlusion (BCCAO) and to investigate the mechanism of 10% hypertonic glycerol neuroprotection. Our suggestion was that 10% glycerol solution infusion could dilate pial arterioles through nitric oxide release and/or stimulation of ATP-sensitive potassium (K(ATP)) channels. Therefore, we studied the effects of hypertonic glycerol after inhibition of nitric oxide synthase, with N(G)-nitro-L-arginine-methyl ester or N(G)-nitro-L-arginine, or K(ATP) channels with glibenclamide.

Geometric characteristics of arterial network of rat pial microcirculation.

Objective: The aim of the study was to assess the geometric characteristics of rat pial microcirculation and describe the vessel bifurcation patterns by 'connectivity matrix'.

Methods: Male Wistar rats were used to visualize pial microcirculation by a fluorescent microscopy technique through an open cranial window, using fluorescein isothiocyanate bound to dextran (molecular weight 70 kDa). The arteriolar network was mapped by stop-frame images.

Protective effects of insulin during ischemia-reperfusion injury in hamster cheek pouch microcirculation.

Objective: The effects of insulin (0.18 nM-0.18 microM) on reduced capillary perfusion, microvascular permeability increase and leukocyte adhesion induced by ischemia-reperfusion injury were investigated in the hamster cheek pouch microcirculation.

Effects of tetraiodothyronine and triiodothyronine on hamster cheek pouch microcirculation.

The aim of the present study was to assess the effects of topically applied triiodothyronine (T(3)) and thyroxine (T(4)) on the arterioles of hamster cheek pouch microcirculation in vivo. Microvessels were visualized using a fluorescent microscopy technique. Topical application of T(3) (3.

Molecular steps involved in light-induced oxidative damage to retinal rods.

Purpose: To define the molecular mechanism underlying light-induced oxidative damage to retinal photoreceptors.

Methods: Oxidative stress was induced in isolated rod photoreceptors by bright 470- to 490-nm light and monitored by measuring the conversion of dihydrorhodamine 123 to rhodamine, with fluorescence microscopy. The effect of the wavelength on oxidant generation was investigated by applying prebleaching stimuli of either 485- or 520-nm light before the bright 470- to 490-nm light.

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.

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.

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).

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.

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.

Inhibition of lipid peroxidation by N-acetylserotonin and its role in retinal physiology.

N-Acetylserotonin (NAS) inhibits the peroxidation of linoleic acid induced by a water-soluble initiator, 2,2'-azobis(2-amidinopropane) (ABAP). Lipid peroxidation was detected by the formation of conjugated dienes, monitored spectrophotometrically at 236 nm. N-Acetylserotonin, at concentrations ranging from 200 nM to 20 microM, reduced the rate of ABAP-initiated lipid peroxidation in a concentration-dependent manner.

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.

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.

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.

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.

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.

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.

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 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.

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.

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.

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.

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.

Intracellular analysis and structural correlates of the organization of inputs to ganglion cells in the retina of the turtle.

Intracellular recording from the ganglion cells of the retina of the turtle Pseudemys scripta elegans shows that the photoresponses are characterized by either of two reversal potentials. These have been related to the bipolar and amacrine cell inputs to ganglion cells. Of the recorded ganglion cells, 33% (type A) are driven predominantly by one type of input, attributed to bipolar cells.