Lourdes Vidal

Ultrastructural and quantitative age-related changes in capillaries of the dorsal lateral geniculate nucleus.

An ultrastructural and quantitative study of age-related changes in the capillaries of the dorsal lateral geniculate nucleus was carried out using male Wistar rats aged 3, 24, and 28 months. The most important structural changes were found in the basal lamina: thickenings either homogeneously distributed or in specific points; spurs towards the astrocyte sheath; and osmiophilic membrane-like inclusions located within the basal lamina. Endothelial cells and pericytes showed an increase in inclusions and dense bodies in the cytoplasm. The quantitative study showed that the most pronounced alteration was the thickening of the basal lamina, which existed at 24 months. Later, at 28 months, thinning of the endothelial cells was observed together with an increase in mitochondria size and the number of pinocytic vesicles. These changes could be an endothelial cell response to compensate for the increasing transport difficulties caused by the thickening of the basal lamina. The progressive age-related changes observed in the structure of the capillaries might have an effect on the regulation of blood and brain tissue exchanges, and thus might contribute to the development of degenerative alterations in surrounding aging neurones.

Reaction of Müller cells in an experimental rat model of increased intraocular pressure following timolol, latanoprost and brimonidine

The aim of this study was to evaluate the reaction of Müller cells in an experimental rat model of intraocular pressure (IOP) and their response to treatment with ocular hypotensive drugs. Episcleral vein cauterization in unilateral eyes of Wistar rats was performed to produce elevated IOP. The animals were divided into five groups: control, experimental, and experimental treated with timolol, latanoprost or brimonidine. Histological sections of retina were studied by immunochemistry with antibodies to glial fibrillary acidic protein (GFAP), and the percentage of labeled area was measured to evaluate the degree of reactive gliosis. In the experimental group, the Müller cells showed hypertrophy and a significant increase in GFAP (4.39+/-0.32%) in relation to retinas of the control group (2.05+/-0.14%). Gliosis was detected in all three treated groups, with a varying increase in GFAP intensity. The timolol-treated group showed the most intense and persistent glial reactivity after 3 months of treatment (13.89+/-0.63%). Treatment with brimonidine, however, resulted in a decrease in the level of GFAP immunoreactivity (8.37+/-0.4%). The group treated with latanoprost showed the lowest glial reactivity (4.8+/-0.36%). Given that all three drugs are effective hypotensive agents, their neuroprotective effect could be related with other factors, such as gliosis, which, over long periods may have noxious effects on the neurons. Thus, hypotensives like brimonidine, and specially latanoprost, may afford greater neuroprotection to the ganglion cells by attenuating the retinal glial reaction.

Nitric oxide synthase in retina and optic nerve head of rat with increased intraocular pressure and effect of timolol

We investigated the expression of nitric oxide synthase (NOS) isoforms -1, -2 and -3 in the retina and optic nerve head (ONH) in an experimental rat model of elevated intraocular pressure (IOP) before and after treatment with timolol, to assess whether its neuroprotective action is associated with the activity of these enzymes. Episcleral vein cauterization in unilateral eyes of Wistar rats was performed to produce elevated IOP. Histological sections of retina and ONH from animals with normal IOP, with elevated IOP, and elevated IOP treated with timolol, were studied by immunohistochemistry with antibodies to NOS-1, NOS-2, and NOS-3. In the control rats, NOS-1 was localized to photoreceptor inner segments, amacrine cells and bipolar cells in the retina, and in astrocytes, pericytes and vascular nitrergic terminals in the ONH. NOS-3 immunostaining localized to the endothelial cells. The rats with elevated IOP showed increased expression of NOS-1 in the plexiform layers of the retina and reactive astrocytes in the ONH. These cells also showed NOS-2 positivity. The rats treated with timolol showed reduced expression of NOS-1 in the retina and ONH. NOS-2 was only detected in a few groups of astrocytes in the ONH. NOS-3 was unchanged in both elevated IOP and timolol-treated groups. These results show that excessive levels of NO synthesized by the NOS-1 and -2 isoforms, considered neurotoxic, might contribute to the progressive lesions of retinal ganglion cell axons. Their reduction after treatment suggests a possible neuroprotective effect of timolol in neurons exposed to excessive amounts of NO.

Quantitative age-related changes in dorsal lateral geniculate nucleus relay neurons of the rat

An ultrastructural and quantitative study of the age-related changes occurring in the relay neurons of the dorsal lateral geniculate nucleus (dLGN) was carried out using male Wistar rats aged 3, 18, 24, and 28 months. Morphometric techniques were used to obtain data regarding cellular activity including soma, nuclear, and nucleolar size. Volume fractions for rough endoplasmic reticulum (RER), mitochondria, and lipofuscin, as well as numbers and sizes of mitochondria and dense bodies (DB) was also calculated. Among the few alterations found in the perikaryon, we can highlight the redistribution and fragmentation of RER and an increase and progressive aggregation of lipofuscin. Quantitative data show a significant decrease in the volume of the soma (-42.77%) and the nucleus (-33.66%), and in the volume fraction of the RER (-18.81%) and mitochondria (-10.16%). A significant increase in lipofuscin (+213.29%), and variations in size and number of mitochondria and dense bodies were also found. Some histophysiological considerations about the findings are discussed. The findings lead to the conclusion that a relative degree of morphological stability is exhibited by relay neurons, although the quantitative data show evident intracellular changes, especially from 24 to 28 months. These changes suggest that accompanying physiological alterations may occur, with putative effects on visual function during ageing.

Quantitative study of the morphological changes in the thyroid gland following IR laser radiation

The morphological changes produced in the thyroid glands of albino rats following radiation with a 904 nm infrared laser were studied. Two different levels of radiation were applied: 46.8 J/cm2 and 140.4 J/cm2. Evaluation of the changes in the densities of the epithelial, colloidal and follicular volumes and of the activation index revealed that the laser beam produced changes in the thyroid parenchyma. It was observed that there was a direct relationship between the severity of the lesion and the radiation energy applied.

Study of the effects of ocular hypotensive drugs on number of neurons in the retinal ganglion layer in a rat experimental glaucoma

Purpose We investigated the effects of antiglaucomatous drugs on neurons in the retinal ganglion layer (RGL) in an experimental model of elevated intraocular pressure (IOP). Methods Three episcleral veins of rats with normal IOP were cauterized. Three months later, we examined the effects on the number of neurons in the RGL as well as in rats submitted to treatment with timolol, latanoprost, or brimonidine. The IOP was measured using a calibrated Tono-Pen XL tonometer before and immediately after cauterization and every 2 weeks for the following 3 months as well as immediately before perfusion. Results The IOP was 14.85±0.65 mmHg in the control group, whereas it was 1.25-fold higher (33.5±1.06 mmHg) in the experimental group. After treatment, the IOP returned to baseline levels. The mean number of neurons per mm2 in the RGL was 33% lower in the experimental group (283±10 cells/mm2) compared with the control group (423±11 cells/mm2). In the groups treated with timolol, latanoprost, or brimonidine, the neuronal loss was less (331±10, 360±15, and 333±3 cells/mm2, respectively), although values did not return to baseline levels. Conclusions This experimental model provokes an immediate, constant, and prolonged increase in IOP and the application of hypotensive agents affords a certain degree of protection to neurons in the RGL.

Quantitative age-related changes in NADPH-diaphorase-positive neurons in the ventral lateral geniculate nucleus

Age-related changes in nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) were examined in the rat ventral lateral geniculate nucleus (vLGN) using histochemical methods. Eighteen rats aged 3, 24, and 26 months were studied using quantitative methods to investigate the number of neurons per mm(2), the cross-sectional area, and the orientation of dendritic processes of NADPH-d-positive neurons. We have described three types of neurons: types A and B are both located in the lateral and medial vLGN (vLGN-l and vLGN-m, respectively), and type C neurons over the optic tract. The number of NADPH-d-positive neurons was significantly reduced in the old rats (-39%) when compared with controls (3-month-old rats). The quantitative analysis of cell areas revealed a significant decrease of somatic size in type B neurons, both in the lateral and medial vLGN, and in C neurons; however, type A neurons did not show significant changes. By quantifying the orientation of dendritic processes, we observed a predominant dorsolateral orientation in type A and B neurons. During aging, there are no changes in the dendritic orientation of neurons located in the vLGN-m; however, vLGN-l neurons show an increase in dendritic processes with dorsoventral orientation. In type C neurons, our results show that 87.4% of dendritic processes are lateromedially oriented at 26 months old. Therefore, the types A and B neurons behave differently during senescence. Type A neurons do not change in size, but those located in the vLGN-l modify the orientation of their dendritic processes; however, type B neurons, reduce their size and those located in the vLGN-l also modify their dendritic process orientation. Finally, the type C neurons modify their size and dendritic process.

Experimental model of ocular hypertension in the rat: study of the optic nerve capillaries and action of hypotensive drugs.

PURPOSE To investigate quantitatively the effect of elevated intraocular pressure (IOP) on the microvasculature of the optic nerve with and without topical treatment with two hypotensive drugs, timolol and latanoprost. METHODS Three groups of rats underwent cauterization of three episcleral veins to produce elevated IOP in the right eye. Two of these groups were treated with timolol or latanoprost for 3 months. Eyeballs were incubated with anti-GLUT-1 polyclonal antibody. GLUT-1-positive capillaries of the optic nerve head (ONH) and optic nerve exit (ON) were examined and analyzed for their number per square millimeter, volume fraction, length per unit volume, surface area per unit volume, and mean diameter. RESULTS An increase in IOP resulted in a significant decrease in microvessel density in the laminar region (LR) and postlaminar region (PR) and ON compared with the control group. The other parameters fell significantly in all regions of the optic nerve. Topical treatment with timolol or latanoprost did not modify the density of the capillaries, although the other parameters increased significantly compared with the untreated experimental group. Additionally, the mean diameter of the capillaries in the LR and the PR recovered after treatment. CONCLUSIONS The results indicate that the capillaries of the LR and the PR of the ONH are the most susceptible to IOP elevation. The authors suggest that timolol and latanoprost have a certain vascular action by increasing the available blood volume, surface area per unit volume, length per unit volume, and diameter of the capillaries of the ONH in these two regions.

Ultrastructural Changes in Thyroid Perifollicular Capillaries During Normal Postnatal Development and After Infrared Laser Radiation

This study was carried out on the perifollicular capillaries of the thyroid gland during postnatal (PN) development in normal conditions and after irradiation (46.8 J/cm2) with an infrared (IR) laser (904 nm). This was done using 11-, 21-, and 35-day-old Wistar rats. The changes in the capillaries were determined using quantitative methods as well as electron microscopy. During normal PN development the most relevant changes were an increase in the size of the capillaries, especially the lumen, thinning of the endothelium and an increase in the size of pinocytotic vesicles. Our results suggest that during PN development, the capillaries undergo some growth and maturation processes until they reach the optimal morphological conditions for their exchange functions. IR laser irradiation seems to stimulate the growth and maturation of endothelial cells in the youngest rats, while in older ones it causes irregular thickening of the endothelium and a reduction of the capillary lumen. These changes could be a sign of functional alterations in follicular cells caused by exposure to IR laser.

Histological study of the thyroid gland following 904-nm laser radiation

The present histological and stereological studies are on thyroid glands of Wistar rats irradiated with an IR laser. The animals were exposed to total doses of 46,80 J/cm2, and sacrificed at 1, 40, and 180 days after the last treatment. A morphological and stereological study was made on follicular epithelium, follicle, stroma, and capillary. The laser beam produced an increase of follicular epithelial volume and hyperplasia of follicular cells in the animals sacrificed at 1, 40, and 180 days after the last treatment. Capillary dilation underwent a progressive decrease as time passed. It was higher in the animals sacrificed after 24 hours, then it decreased after 40 days and finally disappeared after 180 days.