Retinal Neuronal Cells Research Articles

Expression of Brain-derived Neurotrophic Factor Gene in Retina Following Vitreous Tap

Mechanical injuries to the retina following vitreous tap are reported to protect photoreceptor cells in a rat model of the retinal degeneration and enhance the survival rate of retinal ganglion cells in the optic nerve transection. Neurotrophic factors are presumably involved in the protective mechanisms. In order to see whether neurotrophic factors are synthesized in the retina, we studied the expression of neurotrophic factors in the retina following vitreous tap in rats. One eye each of 20 mature rats received transscleral vitreous taps at three points of entry and retinal injury. The retinas were removed and examined at day 0 to 14 of treatment. Following injury, the retina showed increased expression of glial fibrillary acidic protein (GFAP) mRNA and brain-derived neurotrophic factor (BDNF) mRNA. There was no enhancement of neurotrophin-3 (NT-3) mRNA when examined by reverse transcription-polymerase chain reaction (RT-PCR). Activated retinal glial cells may produce BDNF which prevents retinal neuronal cell damage following injury.

New role for the primate fovea: a retinal excavation determines photoreceptor deployment and shape.

In humans, an increasing density of foveal cone photoreceptors occurs slowly over several years after birth, and accounts for a region that subserves high visual acuity. Concurrently, inner retinal cells move centrifugally away from the foveal center. Such developmental rearrangements reflect complex cellular remodeling after the retinal neuronal cells have differentiated and have formed synapses. Explaining foveal morphogenesis is difficult, because differentiated neuronal cells seem incapable of moving actively. Presented here is a biomechanical explanation of how the above events occur. This hypothesis assumes that the cellular movements throughout the retinal layers occur passively as the eye grows and the retina is stretched. Retinal stretch was simulated using virtual engineering models that were analyzed with finite element analysis. A pit combined with retinal stretch causes the retinal layers to deform in a way that accounts for both the centrifugal and centripetal movement of various retinal cell types. Axially directed, tensile forces associated with stretching the retinal tissue surrounding the pit also accounts for the elongated morphology of foveal cone photoreceptors. These simulations suggest that a pit is required for both the centripetal displacement of cone cells toward the center of the fovea, and for the elongated foveal cone morphology. Since the primate fovea may have minimal impact on acuity, its primary role may be to initiate foveal morphogenesis in slowly developing eyes.

Requirement for nitric oxide in retinal neuronal cell death induced by activated Müller glial cells.

Retinal Müller glial cells express the inducible isoform (-2) of nitric oxide (NO) synthase (NOS) in vitro after stimulation by lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) or in vivo in some retinal pathologies. Because NO may have beneficial or detrimental effects in the retina, we have used cocultures of retinal neurons with retinal Müller glial (RMG) cells from mice disrupted for the gene of NOS-2 [NOS-2 (-/-)] to clarify the role of NO in retinal neurotoxicity. We first demonstrated that NO produced by activated RMG cells was not toxic for RMG cells themselves. Second, the NO released from LPS/IFN-gamma-stimulated RMG cells induced neuronal cell death, because no neuronal cell death has been observed in cocultures with RMG cells from NOS-2 (-/-) mice and because inhibition of NOS-2 induction by transforming growth factor-beta or blockade of NO release by different NOS inhibitors prevented neuronal cell death. Addition of urate, a peroxynitrite scavenger, or superoxide dismutase partially prevented neuronal cell death induced by NO, whereas the presence of a poly(ADP-ribose) synthetase inhibitor, caspase inhibitors, or a guanylate cyclase inhibitor had no significant effect on cell death. These results demonstrated that a large release of NO from RMG cells is responsible for retinal neuronal cell death in vitro, suggesting a neurotoxic role for NO and peroxynitrite during retinal inflammatory or degenerative diseases, where RMG cells were activated.

Evaluation of the role of human retinal vascular endothelial cells in the pathogenesis of CMV retinitis

Previous studies of cytomegalovirus (CMV) retinitis have failed to definitively explain the exact mechanism by which CMV gains access to and initiates infection in the retina. Proposed theories have included leakage of the virus through vessels with altered permeability, with subsequent infection of surrounding glial cells. In an attempt to shed further light on this subject, a histopathologic examination of 30 autopsy eyes from patients with known systemic CMV disease was carried out using light microscopy, immunohistochemical and immunofluorescent techniques, and in-situ hybridization. Dual-staining methods were used to identify the exact cell type showing the presence of CMV antigens, namely vascular endothelial cells, glial cells, neuronal cells, and/or leukocytes. In those eyes with CMV retinitis, the sites of full-thickness retinal necrosis revealed viral presence mostly within Müller cells and perivascular glial cells, with focal areas of positive staining within retinal pigment epithelial cells (RPE) and neuronal cells. The retinal capillaries were devoid of endothelial cells in these areas. Adjacent to regions of full-thickness necrosis, some vessels showed the presence of a viral antigen within the endothelial cells. These findings suggest that retinal vascular endothelial cells can be infected with CMV. It can further be hypothesized that infection of vascular endothelial cells leads to infection of the surrounding glial and neuronal cells, with eventual spread to the RPE. Endothelial cells might not be present in areas of full-thickness necrosis due to mechanical forces from adjacent blood flow resulting in the sloughing of these cells.

Expression and Cellular Localization of the Kallikrein–Kinin System in Human Ocular Tissues

Tissue kallikrein is a serine proteinase which processes kininogens to release bioactive kinins. Kinins mediate a variety of biological processes through the interaction with kinin receptors. Kinins are involved in the regulation of blood pressure and local blood flow, vasodilation, smooth muscle contraction and relaxation, production of pain and inflammation, and stimulation of cell proliferation. The tissue kallikrein–kinin system has been implicated in a number of pathophysiological processes such as hypertension, allergy and diabetes mellitus. In the present study, we have identified the expression and localization of components of the kallikrein–kinin system in the human eye by reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses, and in situ hybridization histochemistry. RT-PCR and Southern blot analyses have detected mRNAs of the key components of the system including tissue kallikrein, low molecular weight kininogen, and bradykinin B 1and B 2receptors at high levels in human retina, choroid and ciliary body, and relatively low levels in the optic nerve. In situ hybridization has identified cellular localization of these four mRNAs in ocular tissues. They are expressed in retinal neuronal cells including the outer nuclear layer, inner nuclear layer and ganglion cell layer. These mRNAs were also identified in endothelial cells of ocular blood vessels, ciliary muscle and lens epithelial cells. The sense riboprobes showed negative staining, which indicates the specificity of the antisense riboprobes. These results suggest that the tissue kallikrein–kinin system is produced endogenously in human ocular tissues. Similar expression patterns of kallikrein, kininogen and kinin receptors indicate that the kallikrein–kinin system may function in an autocrine or paracrine fashion in the eye.

Cloning, expression, and tissue distribution of a fifth melanocortin receptor subtype

The melanocortin (MC) peptides mediate a diverse spectrum of biological activities in both the central nervous system and peripheral tissues by interacting with specific guanine nucleotide binding (G protein)-coupled receptors. Previously, four human melanocortin receptor subtypes have been cloned and characterized. In this study, we have isolated mouse complementary DNA (cDNA) and human genomic clones encoding a fifth melanocortin receptor subtype, MC5. Melanocortin peptide stimulation of human MC5, transiently expressed in COS1 cells, results in activation of adenylate cyclase with the following rank order of potency: [Nle4, D-Phe7]-alpha-MSH (melanocyte stimulating hormone) > ACTH (1-24) (adrenocorticotropic hormone) > alpha-MSH > beta-MSH > gamma-MSH. Northern blot hybridization, ribonuclease protection, and reverse transcription/polymerase chain reaction assays indicate that mouse MC5 mRNA is most abundant in skeletal muscle and brain. Lower but detectable levels of MC5 mRNA are also found in RT2-2 retinal neuronal cells, lung, testis, spleen, heart, kidney, and liver.

Immortalized Neuronal and Neuroendocrine Cell Lines by Targeted Oncogenesis in Transgenic Mice Using the PNMT Promoter

Phenylethanolamine N-methyltransferase (PNMT) is the terminal enzyme in the catecholamine biosynthetic pathway, converting norepinephrine to epinephrine. In transgenic mice, 2 kb of the human PNMT promoter directs the expression of the simian virus 40 (SV4O) early region in three classes of retinal neurons and in chromaffin cells of the adrenal medulla. These transgenic animals develop retinal and adrenal medullary tumors between 3 and 4 months of age. We have adapted these tumor cells to cell culture and have derived immortalized retinal neuronal and adrenal chromaffin cell lines. These cell lines express a number of cell- and tissue-specific markers indicative of the differentiated cells of origin.

DEVELOPMENT AND REGENERATION OF RETINAL NEURONS FROM PIGMENT EPITHELIUM IN FROGS -IMPLICATIONS FOR HUMANS?

After injury, frog and tadpole retinal pigment epithelium (RPE) cells are able to differentiate into retinal neuronal cells in a reparative process. In this study, the authors examined …