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  • 1
    Electronic Resource
    Electronic Resource
    Time-course expression of vascular endothelial growth factor as related to the development of the retinochoroidal vasculature in rats (1998)
    Springer
    Experimental brain research 118 (1998), S. 155-160 
    Details
    ISSN: 1432-1106
    Keywords: Key words Retinochoroidal vasculature ; Retinal pigment epithelium ; Vascular endothelial growth factor ; In situ hybridization ; Immunohistochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Growth factors involved in angiogenesis are critical to both the normal and pathological vascular development in the retina and choroid. In the present experiment, the relationship between the vascular endothelial growth factor (VEGF) expression and the retinochoroidal vasculogenesis in Sprague-Dawley rats was investigated using in situ hybridization and immunohistochemistry. It was found that VEGF was produced mainly by astrocytes and Müller cells in the neural retina, and this was correlated temporally and spatially with the retinal vasculogenesis. In addition, it was observed that, although the VEGF expression in the retinal pigment epithelium (RPE) decreased with increasing age, it persisted from the embryonic stage to adulthood. These findings indicate that the VEGF expression in RPE may play a role in the development of the choroidal vessels as well as in the maintenance of the normal structure and permeability of the choriocapillaris in adults.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002210050267
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  • 2
    Electronic Resource
    Electronic Resource
    In situ hybridization analysis of substance P receptor in the rat retina (1996)
    Springer
    Experimental brain research 112 (1996), S. 181-186 
    Details
    ISSN: 1432-1106
    Keywords: Tachykinin receptor ; Retrograde cell labeling ; Optic nerve crush ; Ganglion cell layer ; Inner nuclear layer ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Substance P receptor is known to provide a principal interface between tachykinin peptides and tachykinin-sensitive cells in retinal circuitry and to produce several physiological functions such as excitation of ganglion cells. We reported results of in situ hybridization analysis of substance P receptor in rat retina using digoxigenin-labeled RNA probes to yield discrete cell labeling. Distinct hybridization signal was present in a great majority of ganglion cells that provide retinal fibers to a central target. It was also present in a subpopulation of amacrine cells. Following optic nerve crush, ganglion cells lost their hybridization signal in a time-dependent manner, while hybridization-positive amacrine cells were persistently seen. From the results, we identified the hybridization message as distinctly localized to two systems, output cells and intrinsic cells in retinal circuitry.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00227636
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  • 3
    Electronic Resource
    Electronic Resource
    Immunolocalization of basic fibroblast growth factor during wound repair in rat retina after laser photocoagulation (1996)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 234 (1996), S. 695-702 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract •Background: Basic fibroblast growth factor (bFGF) stimulates the mitogenesis of various cells and plays a key role in wound repair. We studied the immunohistochemical localization of bFGF during wound repair in the rat retina after laser photocoagulation. • Methods: Krypton laser photocoagulation was performed on the eyes of pigmented rats. The eyes were enucleated on days 1, 3, 7, 14 and 28 after the photocoagulation, and the immunohistochemical localization of bFGF was assessed. Two different monoclonal antibodies and one polyclonal antibody against bFGF as first antibodies were used. • Results: Marked immunoreactivity for bFGF was found in the ganglion cell layer, and weak immunoreactivity for bFGF was found in the retinal pigment epithelial (RPE) cells of the normal adult rat retina. On day 3 after laser photocoagulation, the nuclei and cytoplasm of proliferating RPE cells at the center of the photocoagulated lesion showed intense bFGF immunoreactivity. The nuclei of RPE cells around the lesion showed intense bFGF immunoreactivity. Macrophages that migrated into the lesion showed positive staining for bFGF. These immunoreactivity decreased with time. Controls (0.05 M Tris-HCl buffer, normal serum, or these same antibodies preabsorbed with bFGF) did not show positive staining. • Conclusion: The finding of an elevated expression of bFGF immunoreactivity in the photocoagulated lesion suggests that bFGF may play a role in wound repair in the rat retina after laser photocoagulation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00292356
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  • 4
    Electronic Resource
    Electronic Resource
    Vascular endothelial growth factor expression in choroidal neovascularization in rats (1997)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 235 (1997), S. 313-319 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: The pathogenesis of choroidal neovascularization is largely unknown. We investigated vascular endothelial growth factor (VEGF) expression in laser-induced choroidal neovascularization (CNV) in rats. Methods: Intense krypton laser photocoagulation was applied to the posterior poles of the eyes of pigmented rats to induce CNV, which was confirmed by fluorescein angiography and histopathology. The eyeballs were enucleated 1, 3, 7, 14 and 28 days after laser photocoagulation. Cryostat sections were prepared for immunofluorescence staining using anti-VEGF and macrophage marker (ED1) antibodies. The posterior segments of eyeballs pooled from photocoagulated and control rats were submitted for immunoprecipitation and immunoblotting by the anti-VEGF antibody, and reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of VEGF mRNA. Results: Very weak immunoreactivity for anti-VEGF antibody was found in the ganglion cell layer, inner nuclear layer, and retinal pigment epithelium (RPE) in the normal retina. In the development of CNV, strong positive staining for anti-VEGF antibody was found in photocoagulated areas in the subretinal space and choroid. Double immunofluorescence staining showed that many cells in lasered lesions were positive both for anti-VEGF and macrophage marker ED1 antibody staining in the early stage of this model. Immunoblots showed a positive band for the VEGF molecule in treated but not control animals. RT-PCR results demonstrated upregulation of VEGF transcripts in the CNV model compared with normal animals. Conclusions: Our findings showed the upregulation of VEGF expression in experimentally induced CNV, where it may be involved in promoting choroidal angiogenesis. Macrophages may be one of the main sources of VEGF in the early stage of the disease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01739641
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  • 5
    Electronic Resource
    Electronic Resource
    Apoptosis of photoreceptor cells in ornithine-induced retinopathy (1998)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 236 (1998), S. 207-212 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  • Background: The intravitreal injection of ornithine produces selective damage to the retinal pigment epithelium (RPE) and results in a loss of RPE, choriocapillaris and photoreceptor cells. To elucidate the mechanism of secondary retinal atrophy, we investigated the presence of apoptotic cells in a rat model of ornithine-induced retinopathy. • Methods: At 6 and 12 h and 1, 2, 4, 7, 14 and 28 days after an intravitreal injection of L-ornithine hydrochloride in rat eyes, we removed the eyes and subjected them to histopathological examination. We detected apoptotic cells by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate digoxigenin nick end labeling (TUNEL) assay, which stains the 3′-OH ends of fragmented DNA. We used electron microscopy to detect the apoptotic cells morphologically. • Results: RPE cells were selectively damaged immediately after ornithine administration. TUNEL-positive photoreceptor cells appeared exclusively in the photoreceptor cell layer 12 h after ornithine administration. The number of TUNEL-positive cells increased throughout the 2 days following the injection, then decreased markedly. TUNEL-positive cells remained until 28 days, when the photoreceptor cells had disappeared. The ganglion cell layer, inner nuclear layer and damaged RPE cells were negative for TUNEL staining during all stages. The electron microscopic study also revealed the pyknotic nuclei of apoptotic photoreceptor cells. • Conclusion: An intravitreal injection of ornithine caused primary damage to the RPE, and subsequently some of the photoreceptor cells revealed apoptosis by TUNEL assay. These findings suggest the dysfunction of the RPE causes photoreceptor cell death according to the intrinsic program of an apoptotic mechanism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004170050066
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  • 6
    Electronic Resource
    Electronic Resource
    Central serous choroidopathy with bullous retinal detachment (1978)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 206 (1978), S. 169-178 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We report here a new type of secondary retinal detachment that has never been clearly defined. The characteristic features of the disease are: (1) prevalence in middle-aged males, (2) bilateral involvement, (3) frequent existence of prodromal lesions that over long periods resemble central serous retinopathy, (4) in the evolution stage, appearance of multiple yellowish white exudative flecks of one-half to one disc in diameter at or near the posterior pole of the fundus, (5) fluorescein studies revealing pronounced leakage of dye from the choroid into the subretinal space at the sites of exudates, (6) retinal detachment of various degrees with shifting subretinal fluid and without tears, (7) no evidence of intraocular inflammation, (8) no filling abnormalities seen in the choroidal fluorescence, (9) no response to medical therapy, including steroids and antibiotics, (10) photocoagulation to leakage sites leading to rapid resolution of retinal detachment; otherwise, spontaneous healing of detachment occurring within about 7–9 months, leaving fibroblastic macular scars and marked visual loss, and (11) no evidence of systemic findings that may be of etiologic significance. From this characteristic clinical picture, the idea of a new clinical entity must be considered. Our findings in 35 eyes from 18 Japanese patients are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00414743
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  • 7
    Electronic Resource
    Electronic Resource
    Immunolocalization of transforming growth factor β during wound repair in rat retina after laser photocoagulation (1997)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 236 (1997), S. 41-46 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract • Background: Scatter photocoagulation induces regression of retinal neovascularization, but the mechanism of its therapeutic effect is incompletely understood. To elucidate the mechanism of therapeutic effect of photocoagulation is the main focus of our research. We have already demonstrated basic fibroblast growth factor (bFGF) immunolocalization during retinal wound repair following laser photocoagulation. Transforming growth factor beta (TGF β) reportedly inhibits endothelial cell growth and bFGF-induced cell proliferation in vitro. In the present study, we evaluated the immunohistochemical localization of TGF-β1 and -β2 during wound repair in the rat retina following laser photocoagulation. • Methods: Krypton laser photocoagulation was performed on the eyes of pigmented rats. The eyes were then enucleated on day 1, 3, 7, 14, 28 or 56 following the photocoagulation and enrolled into the analysis of immunohistochemical localization of TGF-β1 and -β2. • Results: Immunoreactivity for TGF-β1 and -β2 was present in the ganglion cell layer and photoreceptor outer segments of the normal adult rat retina. The cytoplasm of RPE cells at the photocoagulated lesion showed intense TGF-β1 and -β2 immunoreactivity on day 3 after laser photocoagulation. Macrophages that migrated into the lesion lacked positive staining for TGF-β1 and -β2. TGF-β immunoreactivity in RPE cells continued to be upregulated for more than 1 month compared with that in normal RPE cells. Controls did not exhibit any positive staining. • Conclusion: An elevated expression of TGF-β immunoreactivity for a longer period of time than bFGF was observed in RPE cells at the photocoagulated lesion in vivo. In the late phase of retinal wound repair, TGF-β may inhibit cell proliferation induced by mitogens, introduce an end stage of cellular events, and induce extracellular matrix induction.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004170050040
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  • 8
    Electronic Resource
    Electronic Resource
    Retinal degeneration induced by N-methyl-N-nitrosourea in Syrian golden hamsters (1998)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 236 (1998), S. 688-695 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  · Background: The sequential retinal changes in Syrian golden hamsters induced by N-methyl-N-nitrosourea (MNU) have not been studied. · Methods: Female hamsters received a single intraperitoneal injection of 90 mg/kg MNU at 50 days of age, and the retina was examined light and electron microscopically, immunohistochemically and by the TdT-mediated dUTP-digoxigenin nick end labeling (TUNEL) method until 20 weeks after the treatment. · Results: The retinal changes were as follows: (1) Photoreceptor apoptosis occurred 1 day after the treatment and resulted in photoreceptor loss at day 7. During the degeneration, Müller cell proliferation was conspicuous at day 5. (2) After the photoreceptor cell loss, migration of the pigment epithelial cells in all layers of the retina which were in contact with blood vessels occurred. Due to the Müller cell proliferation, gliosis was prominent at the later stage. · Conclusions: The MNU injection caused photoreceptor apoptosis followed by pigment epithelial cell migration around the blood vessels, accompanied by gliosis. The primary event and the course of this disease closely resemble those of retinitis pigmentosa in humans.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004170050142
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  • 9
    Electronic Resource
    Electronic Resource
    Transfection of basic fibroblast growth factor (bFGF) gene or bFGF antisense fene into human fetinal pigment epithelial cells (1999)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 237 (1999), S. 678-684 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  · Background: Transplantation of RPE cells offers a potential of restoring retinal pigment epithelium (RPE) function and has been shown to be effective in the dystrophic RCS rat model. Recently, RPE transplantation was attempted in patients with age-related macular degeneration. Basic fibroblast growth factor (bFGF) plays important roles in maintaining normal retinal function. The purpose of this study was to introduce bFGF sense or antisense cDNA into human RPE cells to alter the expression of bFGF. · Methods: Human bFGF sense cDNA or antisense cDNA was inserted into the pBK-CMV vector. For stable gene expression, we introduced the plasmids into RPE cells using the electroporation method. Following electroporation, transfected RPE cells were cultured and resistant cells were selected in the presence of antibiotic G418. We analyzed the expression of the transfected genes in the cloned RPE cells by polymerase chain reaction (PCR) and by reverse transcription (RT)-PCR. · Results: Cloned RPE cells in which the bFGF sense or antisense cDNA had been efficiently transfected were established. PCR and RT-PCR analysis demonstrated not only the presence but also the expression of bFGF sense or antisense cDNA in the transfected RPE cells. · Conclusions: Human bFGF sense cDNA or antisense cDNA can be efficiently introduced into cultured RPE cells by the electroporation method. The successful expression of the genes into RPE cells demonstrated that this technique can be used to regulate bFGF expression and thus increase the scope of RPE transplantation for the treatment of retinal diseases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004170050296
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  • 10
    Electronic Resource
    Electronic Resource
    Expression of basic fibroblast growth factor and its receptor mRNA in retinal tissue following ischemic injury in the rat (1998)
    Springer
    Graefe's archive for clinical and experimental ophthalmology 236 (1998), S. 295-300 
    Details
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  · Background: Our purpose was to determine the time-dependent changes of expression of basic fibroblast growth factor (bFGF) and its receptor in pressure-induced retinal ischemia in rats. · Methods: Retinal ischemia was induced in Wistar rats by increasing the intraocular pressure to 110 mmHg for 45 min by cannulation into the eyes. At the end of the ischemic period, reperfusion of the retinal vasculature was confirmed. Localization of bFGF and FGF receptor-1 (FGF-R) mRNAs were evaluated by in situ hybridization at various times after reperfusion. The reverse-transcription polymerase chain reaction (RT-PCR) method was used to detect the expression of bFGF and FGF-R mRNA in the sensory retina. · Results: In normal sensory retina, bFGF and FGF-R mRNAs were observed in the ganglion cell layer and inner nuclear layer. bFGF gene expression in the sensory retina increased within 24 h, particularly at 6–12 h. FGF-R gene expression increased earlier than that of bFGF. By RT-PCR, expression of bFGF gene reached a peak at 6–24 h, and FGF-R reached a peak at 3–12 h. These RT-PCR results are comparable to those of in situ hybridization. · Conclusions: These results demonstrate that transient retinal ischemia leads to the induction of bFGF mRNA synthesis, and suggest that bFGF has a protective role, e.g., a defense mechanism for the sensory retina.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004170050081
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