Angiogênese e doenças da retina / Angiogenesis and retinal diseases

Angiogênese é o processo de formação de vasos sangüíneos a partir de vasos preexistentes, que ocorre em condições fisiológicas e patológicas. É fenômeno complexo no qual participam inúmeras moléculas que estimulam e inibem a formação dos neovasos. O aumento da permeabilidade vascular e a neovascularização sub-retiniana são as causas da perda visual nas doenças proliferativas da retina, como a degeneração macular relacionada à idade e a retinopatia diabética, e o fator de crescimento do endotélio vascular ("vascular endothelial growth factor", VEGF) desempenha um papel muito importante nesse processo. Existem quatro isoformas da molécula de VEGF biologicamente ativas em seres humanos, das quais o VEGF165 é a isoforma predominante no olho humano, e existem evidências de que seja a isoforma responsável pela neovascularização patogênica no olho. Além de ser potente mitógeno de células endoteliais, o VEGF aumenta a permeabilidade vascular, inibe a apoptose das células endoteliais e promove migração de precursores de células endoteliais. O VEGF não é a única molécula cuja expressão está aumentada na angiogênese patológica. O fator de crescimento de fibroblasto ("basic fibroblast growth factor", bFGF), as angiopoetinas, o fator derivado do epitélio pigmentado ("pigment epithelium-derived factor", PEDF) e os fatores de adesão relacionados à matriz extracelular também exercem papel importante no balanço entre fatores pró- e antiangiogênicos. Todo o conhecimento adquirido sobre o mecanismo da angiogênese ocular patológica tem possibilitado o desenvolvimento de vários inibidores desse processo. Atualmente existem dois anticorpos anti-VEGF para uso intravítreo e outras abordagens terapêuticas do bloqueio da angiogênese ocular estão em fase de desenvolvimento. As novas drogas deverão ser armas poderosas no tratamento das principais causas de cegueira legal irreversível em indivíduos com mais de 65 anos.

Angiogenesis is the process involving the growth of new blood vessels from preexisting vessels which occurs in both physiologic and pathological settings. It is a complex process controlled by a large number of modulating factors, the pro-and antiangiogenic factors. The underlying cause of vision loss in proliferative retinal diseases, such as age-related macular degeneration and proliferative diabetic retinopathy, are increased vascular permeability and choroidal neovascularization, and vascular endothelial growth factor (VEGF) plays a central role in this process. VEGF is produced in the eye by retinal pigment epithelium (RPE) cells and is upregulated by hypoxia. There are four major biologically active human isoforms, of which VEGF165 is the predominant in the human eye and appears to be the responsible for pathological ocular neovascularization. Besides being a potent and specific mitogen for endothelial cells, VEGF increases vascular permeability, inhibits endothelial cells apoptosis, and is a chemoattractant for endothelial cell precursors. VEGF is not the only growth factor involved in ocular neovascularization. Basic fibroblast growth factor (bFGF), angiopoietins, pigment epithelium-derived factor (PEDF), and adhesion molecules also play a role in the pro- and antiangiogenic balance. Advances in the understanding of the bases of pathological ocular angiogenesis and identification of angiogenesis regulators have enabled the development of novel therapeutic agents. Anti-VEGF antibodies have been developed for intravitreal use, and other approaches are currently under investigation. These new drugs may be powerful tools for the treatment of the leading causes of irreversible blindness in people over age 65.

Effect of the absence of heat shock protein 70.1 (hsp70.1) on retinal photoreceptors in normal and rd mice

The purposes of this study are to elucidate the retinal changes of heat shock protein 70.1 (hsp70.1) knockout mice and to compare them between in normal and in retinal degeneration (rd) mice. Eyes of hsp70.1 wild type (+/+) and knockout (-/-) mice in the C57BL/6 or FVB genetic backgrounds respectively, which were reared in the normal environment, were examined by fundus photography, electroretinography, light microscopy, terminal dUTP nick-end labeling (TUNEL) stain, and immunohistochemistry. In C57BL/6 mice, fundus photography showed no changes between hsp70.1+/+ and -/- mice at 1 and 6 months of age. Electroretinographic examination showed a tendency of decreased amplitude of a- and b-wave with aging in both genotype, but there were not different statistically. The ratios of the thickness of inner nuclear and outer nuclear layer to the retinal thickness were respectively decreased with aging in both genotype, but there were not different statstically. TUNEL assay showed a few positively labeled cells in the ganglion cell, inner nuclear and outer nuclear layers and the immunohistochemistry showed no immunopositivity of hsp70 in the inner segments of photoreceptor cell layer in both genotype. In rd mice, fundus photography showed a narrowing of the retinal vessels at the age of 4 weeks, however, there were no differences of retinal changes including pigment epithelial layer in both genotype. Electroretinographic examination at the postnatal 2, 3 and 4 weeks showed no differences between them. Loss of photoreceptor cell and outer nuclear layers showed no differences in both genotype. In conclusion, there were no differences of the retinal changes at least under the normal environmental condition in hsp70.1+/+ and -/- mice. These results show that hsp70.1-/- mice can be used to study the role of hsp70.1 to the external stress to the retina.

Death in dish: controls of apoptosis within the developing retinal tissue

Studies of programmed cell death in the developing retina in vitro are currently reviewed. The results of inhibiting protein synthesis in retinal explants indicate two mechanisms of apoptosis. One mechanism depends on the synthesis of positive modulators ('killer proteins'), while a distinct, latent mechanism appears to be continuously blocked by negative modulators. Extracellular modulators of apoptosis include the neurotrophic factors NT-4 and BDNF, while glutamate may have either a positive or a negative modulatory action on apoptosis. Several protein kinases selectively modulate apoptosis in distinct retinal layers. Calcium and nitric oxide were also shown to affect apoptosis in the developing retianl tissue. The protein c-Jun was found associated with apoptosis in various circumstances, while p53 seems to be selectively expressed in some instances of apoptosis. The results indicate that the sensitivity of each retinal cell to apoptosis is controlled by multiple, interactive, cell type- and context-specific mechanisms. Apoptosis in the retina depends on a critical interplay of extracellular signals delivered through neurotrophic factors, neurotransmitters and neuromodulators, several signal transduction pathways, and the expression of a variety of genes.

Calretinin in the mouse superior colliculus originates from retinal ganglion cells

We investigated the origin of the calretinin-immunoreactive fibers in the mouse superior colliculus. The dense plexus of calretinin-positive fibers in the superficial layers of the colliculus was completely eliminated after eye enucleation. Retrograde tracing combined with immunohistochemistry revealed many calretinin-positive small-to-medium retinal ganglion cells projecting to the colliculus. These results indicate that calretinin-containing ganglion cells are the source of this calcium-binding protein in the superficial layers of the superior colliculus.