Improved semiautomatic method for morphometry of angiogenesis and lymphangiogenesis in corneal flatmounts

Bock, Felix; Onderka, Jasmine; Hos, Deniz; Horn, Folkert K.; Martus, Peter; Cursiefen, Claus

doi:10.1016/j.exer.2008.08.007

Cited by 83 publications

(83 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The areas covered with vessels were then analyzed with an algorithm established in Cell^F software (Olympus, Hamburg, Germany) as previously described. 42 Briefly, gray-scale images of micrographs were generated to optimize vessel signal to noise ratio and the corneal area was subsequently defined along the limbus for quantitative analysis of blood and lymphatic vessels. The area covered by blood and lymphatic vessels in the cornea was correlated with the total area of the cornea (vessel ratio).…”

Section: Corneal Vessel Area Quantitationmentioning

confidence: 99%

Involvement of Corneal Lymphangiogenesis in a Mouse Model of Allergic Eye Disease

Lee

Hos

Blanco

et al. 2015

Invest. Ophthalmol. Vis. Sci.

View full text Add to dashboard Cite

PURPOSE. The contribution of lymphangiogenesis (LA) to allergy has received considerable attention and therapeutic inhibition of this process via targeting VEGF has been considered. Likewise, certain inflammatory settings affecting the ocular mucosa can trigger pathogenic LA in the naturally avascular cornea. Chronic inflammation in allergic eye disease (AED) impacts the conjunctiva and cornea, leading to sight threatening conditions. However, whether corneal LA is involved is completely unknown. We addressed this using a validated mouse model of AED.METHODS. Allergic eye disease was induced by ovalbumin (OVA) immunization and chronic OVA exposure. Confocal microscopy of LYVE-1-stained cornea allowed evaluation of corneal LA, and qRT-PCR was used to evaluate expression of VEGF-C, -D, and -R3 in these mice. Administration of VEGF receptor (R) inhibitor was incorporated to inhibit corneal LA in AED. Immune responses were evaluated by in vitro OVA recall responses of T cells, and IgE levels in the serum. RESULTS.Confocal microscopy of LYVE-1-stained cornea revealed the distinct presence of corneal LA in AED, and corroborated by increased corneal expression of VEGF-C, -D, and -R3. Importantly, prevention of corneal LA in AED via VEGFR inhibition was associated with decreased T helper two responses and IgE production. Furthermore, VEGFR inhibition led a significant reduction in clinical signs of AED.CONCLUSIONS. Collectively, these data reveal that there is a distinct involvement of corneal LA in AED. Furthermore, VEGFR inhibition prevents corneal LA and consequent immune responses in AED.

show abstract

Section: Corneal Vessel Area Quantitationmentioning

confidence: 99%

Involvement of Corneal Lymphangiogenesis in a Mouse Model of Allergic Eye Disease

Lee

Hos

Blanco

et al. 2015

Invest. Ophthalmol. Vis. Sci.

View full text Add to dashboard Cite

show abstract

“…Vascularized area and total corneal area were measured using ImageJ software with a method similar to that of Bock et al (2008). Filters were applied to subtract background, reduce noise, and enhance contrast.…”

Section: Dce/dcementioning

confidence: 99%

A Pathogenic Relationship Between a Regulator of the Actin Cytoskeleton and Serum Response Factor

et al. 2010

View full text Add to dashboard Cite

Cell hyperproliferation, inflammation, and angiogenesis are biological processes central to the pathogenesis of corneal disease, as well as other conditions including tumorigenesis and chronic inflammatory disorders. Due to the number of disease conditions that arise as a result of these abnormalities, identifying the molecular mechanisms underlying these processes is critical. The avascular and transparent cornea serves as a good in vivo model to study the pathogenesis of cell hyperproliferation, inflammation, and angiogenesis. Corneal disease 1 (Dstn corn1 ) mice are homozygous for a spontaneous null allele of the destrin (Dstn) gene, which is also known as actin depolymerizing factor (ADF). These mice exhibit abnormalities in the cornea including epithelial cell hyperproliferation, stromal inflammation, and neovascularization. We previously identified that the transcription factor, serum response factor (SRF) and a number of its target genes are upregulated in the cornea of these mice. In this study, we show that conditional ablation of Srf in the corneal epithelium of a diseased Dstn corn1 cornea results in the rescue of the epithelial cell hyperproliferation, inflammation, and neovascularization phenotypes, delineating an epithelial cell-specific role for SRF in the development of all of these abnormalities. Our study also demonstrates that Dstn is genetically upstream of Srf and defines a new functional role for SRF as the master regulator of a hyperproliferative, inflammatory phenotype accompanied by neovascularization.

show abstract

“…24 Lymphatic vessels were detected by threshold setting including the bright vessels and excluding the dark background as described previously. 24 …”

Section: Neovascularization Areamentioning

confidence: 99%

Genetic Heterogeneity of Lymphangiogenesis in Different Mouse Strains

Regenfuß

Onderka

Bock

et al. 2010

The American Journal of Pathology

View full text Add to dashboard Cite

Lymphangiogenesis plays an important role in tumor metastasis, wound healing, and immune reactions, such as after organ transplantation. Furthermore, novel antilymphangiogenic drugs are moving into clinical medicine, but so far nothing is known about a potential genetic heterogeneity influencing lymphangiogenesis. Using the mouse cornea micropocket assay (VEGF-C) and the suture-induced corneal neovascularization model in different inbred and wild-derived mouse strains (Balb/cAnNCrl, C57BL/6NCrl, 129S1/SvImJ, SJL/ JCrl, Cast/EiJ, FVB/NCrl), significant differences in the lymphangiogenic response were detected: the lymphvascularized area varied up to 1.9-fold in the micropocket assay and up to 1.7-fold in the suture-induced neovascularization model between the "low-responder" strain BALB/c and the "high-responder" strain FVB in response to the same stimulus. Furthermore, the number of physiological lymphatic vascular extensions into the marginal zone of the normally alymphatic cornea in untreated eyes again showed a difference of 1.6-fold between low-and high-responders. An anti-inflammatory (prednisolone acetate) and a specific anti(lymph)angiogenic therapy (blocking anti-VEGFR-3 antibody) had different effects on the lymphvascularized area in BALB/c mice and FVB mice, suggesting a different responsiveness to antilymphangiogenic treatments. These data for the first time demonstrate significant differences in the lymphangiogenic response of several mouse strains and suggest underlying genetic factors influencing the lymphangiogenic response. These considerations need to be taken into account when using different mouse strains to study lymphangiogenesis and may also explain different success of antilymphangiogenic treatments in tumor patients.

show abstract

Improved semiautomatic method for morphometry of angiogenesis and lymphangiogenesis in corneal flatmounts

Cited by 83 publications

References 14 publications

Involvement of Corneal Lymphangiogenesis in a Mouse Model of Allergic Eye Disease

Involvement of Corneal Lymphangiogenesis in a Mouse Model of Allergic Eye Disease

A Pathogenic Relationship Between a Regulator of the Actin Cytoskeleton and Serum Response Factor

Genetic Heterogeneity of Lymphangiogenesis in Different Mouse Strains

Contact Info

Product

Resources

About