Evolution of neovascularization in mice with overexpression of vascular endothelial growth factor in photoreceptors

Takao Tobe, Naoyuki Okamoto, Melissa A. Vinores, Nancy L. Derevjanik, Stanley A. Vinores, Donald J Zack, Peter A Campochiaro

Research output: Contribution to journalArticle

Abstract

PURPOSE. To determine the earliest changes that occur in the retina after the onset of ectopic expression of vascular endothelial growth factor (VEGF) by photoreceptors in transgenic mice, to characterize the development of neovascularization (NV), and to determine the feasibility of using these mice to test the efficacy of antiangiogenic agents. METHODS. The time course of expression of VEGF transgene mRNA was determined by reverse transcription- polymerase chain reaction (RT-PCR). Histopathologic changes in the retina were investigated by light and electron microscopy and immunocytochemistry. Standard and confocal fluorescence microscopy and image analysis were used to evaluate NV in retinal whole mounts. RESULTS. VEGF transgene mRNA was first detected in the retina by RT-PCR on postnatal day 6 (P6) and increased over the next several days to reach a constant steady-state level between P14 and P21. Abnormal cells were seen in the outer nuclear layer on P10 and among photoreceptors on P14; by P18 there were cell aggregates in the subretinal space with evidence of lumen formation. The invading cells were demonstrated to be endothelial cells by staining with an endothelial cell-specific lectin. Whole mounts of retinas perfused with fluorescein-labeled dextran showed a similar sequence of events, with sprouts from retinal vessels in the deep capillary bed seen on P14 and vessels reaching the subretinal space by P18. Confocal and standard fluorescence microscopy and changes in the number and area of neovascular lesions in the subretinal space over time measured by image analysis suggest gradual enlargement and coalescence of vascular complexes. The subretinal NV was progressively engulfed by the retinal pigmented epithelium. Invasion of blood vessels from the choroid was not identified in any specimen. CONCLUSIONS. These data support the feasibility of using rhodopsin-VEGF transgenic mice to study tissue-specific aspects of NV in the retina and to test antiangiogenic agents for inhibition of intraretinal and subretinal NV.

Original languageEnglish (US)
Pages (from-to)180-188
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume39
Issue number1
StatePublished - Jan 1998

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Vascular Endothelial Growth Factor A
Retina
Angiogenesis Inhibitors
Transgenes
Fluorescence Microscopy
Transgenic Mice
Reverse Transcription
Blood Vessels
Endothelial Cells
Retinal Neovascularization
Retinal Vessels
Polymerase Chain Reaction
Messenger RNA
Rhodopsin
Choroid
Lectins
Confocal Microscopy
Electron Microscopy
Epithelium
Immunohistochemistry

ASJC Scopus subject areas

  • Ophthalmology

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Evolution of neovascularization in mice with overexpression of vascular endothelial growth factor in photoreceptors. / Tobe, Takao; Okamoto, Naoyuki; Vinores, Melissa A.; Derevjanik, Nancy L.; Vinores, Stanley A.; Zack, Donald J; Campochiaro, Peter A.

In: Investigative Ophthalmology and Visual Science, Vol. 39, No. 1, 01.1998, p. 180-188.

Research output: Contribution to journalArticle

Tobe, Takao ; Okamoto, Naoyuki ; Vinores, Melissa A. ; Derevjanik, Nancy L. ; Vinores, Stanley A. ; Zack, Donald J ; Campochiaro, Peter A. / Evolution of neovascularization in mice with overexpression of vascular endothelial growth factor in photoreceptors. In: Investigative Ophthalmology and Visual Science. 1998 ; Vol. 39, No. 1. pp. 180-188.
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AU - Vinores, Melissa A.

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AU - Zack, Donald J

AU - Campochiaro, Peter A

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N2 - PURPOSE. To determine the earliest changes that occur in the retina after the onset of ectopic expression of vascular endothelial growth factor (VEGF) by photoreceptors in transgenic mice, to characterize the development of neovascularization (NV), and to determine the feasibility of using these mice to test the efficacy of antiangiogenic agents. METHODS. The time course of expression of VEGF transgene mRNA was determined by reverse transcription- polymerase chain reaction (RT-PCR). Histopathologic changes in the retina were investigated by light and electron microscopy and immunocytochemistry. Standard and confocal fluorescence microscopy and image analysis were used to evaluate NV in retinal whole mounts. RESULTS. VEGF transgene mRNA was first detected in the retina by RT-PCR on postnatal day 6 (P6) and increased over the next several days to reach a constant steady-state level between P14 and P21. Abnormal cells were seen in the outer nuclear layer on P10 and among photoreceptors on P14; by P18 there were cell aggregates in the subretinal space with evidence of lumen formation. The invading cells were demonstrated to be endothelial cells by staining with an endothelial cell-specific lectin. Whole mounts of retinas perfused with fluorescein-labeled dextran showed a similar sequence of events, with sprouts from retinal vessels in the deep capillary bed seen on P14 and vessels reaching the subretinal space by P18. Confocal and standard fluorescence microscopy and changes in the number and area of neovascular lesions in the subretinal space over time measured by image analysis suggest gradual enlargement and coalescence of vascular complexes. The subretinal NV was progressively engulfed by the retinal pigmented epithelium. Invasion of blood vessels from the choroid was not identified in any specimen. CONCLUSIONS. These data support the feasibility of using rhodopsin-VEGF transgenic mice to study tissue-specific aspects of NV in the retina and to test antiangiogenic agents for inhibition of intraretinal and subretinal NV.

AB - PURPOSE. To determine the earliest changes that occur in the retina after the onset of ectopic expression of vascular endothelial growth factor (VEGF) by photoreceptors in transgenic mice, to characterize the development of neovascularization (NV), and to determine the feasibility of using these mice to test the efficacy of antiangiogenic agents. METHODS. The time course of expression of VEGF transgene mRNA was determined by reverse transcription- polymerase chain reaction (RT-PCR). Histopathologic changes in the retina were investigated by light and electron microscopy and immunocytochemistry. Standard and confocal fluorescence microscopy and image analysis were used to evaluate NV in retinal whole mounts. RESULTS. VEGF transgene mRNA was first detected in the retina by RT-PCR on postnatal day 6 (P6) and increased over the next several days to reach a constant steady-state level between P14 and P21. Abnormal cells were seen in the outer nuclear layer on P10 and among photoreceptors on P14; by P18 there were cell aggregates in the subretinal space with evidence of lumen formation. The invading cells were demonstrated to be endothelial cells by staining with an endothelial cell-specific lectin. Whole mounts of retinas perfused with fluorescein-labeled dextran showed a similar sequence of events, with sprouts from retinal vessels in the deep capillary bed seen on P14 and vessels reaching the subretinal space by P18. Confocal and standard fluorescence microscopy and changes in the number and area of neovascular lesions in the subretinal space over time measured by image analysis suggest gradual enlargement and coalescence of vascular complexes. The subretinal NV was progressively engulfed by the retinal pigmented epithelium. Invasion of blood vessels from the choroid was not identified in any specimen. CONCLUSIONS. These data support the feasibility of using rhodopsin-VEGF transgenic mice to study tissue-specific aspects of NV in the retina and to test antiangiogenic agents for inhibition of intraretinal and subretinal NV.

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