Visualization of lymphatic vessels by Prox1-promoter directed GFP reporter in a bacterial artificial chromosome-based transgenic mouse

Choi, Inho; Chung, Hee Kyoung; Ramu, Swapnika; Lee, Ha Neul; Kim, Kyu Eui; Lee, Sun-Ju; Yoo, Jae Gyu; Choi, Dongwon; Lee, Yong Suk; Aguilar, Berenice; Hong, Young‐Kwon

doi:10.1182/blood-2010-07-298562

Cited by 214 publications

(244 citation statements)

References 26 publications

Supporting

Mentioning

239

Contrasting

Order By: Relevance

“…This down-regulation may account for the low expression levels of the EGFPLuc reporter in adult mice. By contrast, in previously described transgenic reporter models in which fluorescent protein expression is driven by Prox1 transcriptional control elements, reporter expression is sustained from embryos to adults (16,17). This may represent an advantage of these models for fluorescence visualization of individual lymphatics in adults.…”

Section: Discussionmentioning

confidence: 74%

“…Several transgenic mouse lines for fluorescent visualization of lymphatic vessels have recently been reported. All these lines are based on gene-targeted BAC transgenic constructs to express either GFP (16), mOrange (17), or Tomato (18) fluorescent proteins under Prox-1 transcriptional control. Positron emission tomography (PET) combined with radiolabeled anti-LYVE-1 antibodies have also been used for in vivo imaging of lymphatics in mice (19).…”

mentioning

confidence: 99%

See 1 more Smart Citation

In vivo imaging of lymphatic vessels in development, wound healing, inflammation, and tumor metastasis

Martínez-Corral

Olmeda

Diéguez-Hurtado

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

107

106

View full text Add to dashboard Cite

Lymphatic vessel growth or lymphangiogenesis occurs during embryonic development and wound healing and plays an important role in tumor metastasis and inflammatory diseases. However, the possibility of noninvasive detection and quantification of lymphangiogenesis has been lacking. Here, we present the Vegfr3 EGFPLuc mouse model, where an EGFP-luciferase fusion protein, expressed under the endogenous transcriptional control of the Vegfr3 gene, allows the monitoring of physiological and pathological lymphangiogenesis in vivo. We show tracking of lymphatic vessel development during embryogenesis as well as lymphangiogenesis induced by specific growth factors, during wound healing and in contact hypersensitivity (CHS) -induced inflammation where we also monitor down-regulation of lymphangiogenesis by the glucocorticoid dexamethasone. Importantly, the Vegfr3-reporter allowed us to tracking tumor-induced lymphangiogenesis at the tumor periphery and in lymph nodes in association with the metastatic process. This is the first reporter mouse model for luminescence imaging of lymphangiogenesis. It should provide an important tool for studying the involvement of lymphangiogenesis in pathological processes.bioluminescence | fluorescence | knockin mouse

show abstract

Section: Discussionmentioning

confidence: 74%

mentioning

confidence: 99%

In vivo imaging of lymphatic vessels in development, wound healing, inflammation, and tumor metastasis

Martínez-Corral

Olmeda

Diéguez-Hurtado

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

107

106

View full text Add to dashboard Cite

show abstract

“…We first examined whether SC is detectable with lymphatic and blood vascular markers in the whole-mounted and cross-sectioned corneas of Prox1-GFP reporter mice (17). We confirmed that Prox1 was highly expressed in the CD31 + SC and limbal LVs and moderately expressed in aqueous veins (AVs), which act as collecting channels between SC and episcleral blood vessels (BVs) (Figure 1, A-D, and Supplemental Video 1; supplemental material available online with this article; doi:10.1172/JCI75392DS1).…”

Section: Resultsmentioning

confidence: 99%

“…Specific pathogen-free C57BL/6J mice, Tie2-GFP (FVB/N) mice, and R26 mTmG/+ mice were purchased from the Jackson Laboratory. Prox1-GFP mice (17), Vegfc +/LacZ mice (34), and Prox1-Cre ERT2 mice (58) were transferred and bred in our pathogen-free animal facilities. Prox1-GFP mice were backcrossed for 6 or more generations to C57BL/6J.…”

Section: Methodsmentioning

confidence: 99%

Lymphatic regulator PROX1 determines Schlemm’s canal integrity and identity

et al. 2014

Self Cite

View full text Add to dashboard Cite

“…Gene-targeted mice expressing a Cre-recombinase or a fluorophore under an LEC-specific promoter have recently been generated, [32][33][34][35][36] but only 2 IVM studies performed in such mice have been reported so far. 35,36 Therefore, to date, LVs have mainly been visualized in IVM by in vivo labeling with intracutaneously injected fluorescent antibodies.…”

Section: Discussionmentioning

confidence: 99%

Differential requirement for ROCK in dendritic cell migration within lymphatic capillaries in steady-state and inflammation

Nitschké

Aebischer

Abadier

et al. 2012

Blood

135

View full text Add to dashboard Cite

Dendritic cell (DC) migration via lymphatic vessels to draining lymph nodes (dLNs) is crucial for the initiation of adaptive immunity. We imaged this process by intravital microscopy (IVM) in the ear skin of transgenic mice bearing redfluorescent vasculature and yellowfluorescent DCs. DCs within lymphatic capillaries were rarely transported by flow, but actively migrated within lymphatics and were significantly faster than in the interstitium. Pharmacologic blockade of the Rho-associated protein kinase (ROCK), which mediates nuclear contraction and de-adhesion from integrin ligands, significantly reduced DC migration from skin to dLNs in steady-state. IVM revealed that ROCK blockade strongly reduced the velocity of interstitial DC migration, but only marginally affected intralymphatic DC migration. By contrast, during tissue inflammation, ROCK blockade profoundly decreased both interstitial and intralymphatic DC migration. Inhibition of intralymphatic migration was paralleled by a strong up-regulation of ICAM-1 in lymphatic endothelium, suggesting that during inflammation ROCK mediates de-adhesion of DC-expressed integrins from lymphatic-expressed ICAM-1. Flow chamber assays confirmed an involvement of lymphatic-expressed ICAM-1 and DC-expressed ROCK in DC crawling on lymphatic endothelium. Overall, our findings further define the role of ROCK in DC migration to dLNs and reveal a differential requirement for ROCK in intralymphatic DC crawling during steadystate and inflammation. (Blood. 2012; 120(11):2249-2258) IntroductionDendritic cells (DCs) are important in the initiation of adaptive immune responses. In peripheral tissues, such as the skin, DCs take up antigen, mature, and migrate via lymphatic vessels (LVs) to draining lymph nodes (dLNs), where they present antigen to resting T cells. Although this migration pattern has been known for more than 20 years, 1 DC migration into LVs is only now starting to be unraveled at the cellular level using live imaging technologies. [2][3][4] Before transmigrating across the lymphatic endothelium, DCs first need to squeeze through preformed, narrow pores present in the lymphatic basement membrane (BM). 3 DC entry into LVs is thought to occur at the level of primary lymphatic capillaries, which feature discontinuous cell junctions with "button"-like accumulations of cell adhesion molecules. 5 At the sites of such buttons, lymphatic endothelial cells (LECs) partially overlap and generate open flaps, through which leukocytes enter into lymphatics. 3,5 The most prominent mediator of DC migration into afferent lymphatics is CCL21, a chemokine constitutively expressed in LVs. 6,7 More recently, also other LEC-expressed molecules that mediate DC migration via LVs to dLNs have been identified. [8][9][10][11] Notably, ICAM-1 and VCAM-1, which are up-regulated in LVs during inflammation, 8,12 have been implicated in this process. 8 Intriguingly, experiments performed with pan-integrin knockout DCs revealed that DC migration to dLNs in steady-state was integrin-independent. 2 This ...

show abstract

Visualization of lymphatic vessels by Prox1-promoter directed GFP reporter in a bacterial artificial chromosome-based transgenic mouse

Cited by 214 publications

References 26 publications

In vivo imaging of lymphatic vessels in development, wound healing, inflammation, and tumor metastasis

In vivo imaging of lymphatic vessels in development, wound healing, inflammation, and tumor metastasis

Lymphatic regulator PROX1 determines Schlemm’s canal integrity and identity

Differential requirement for ROCK in dendritic cell migration within lymphatic capillaries in steady-state and inflammation

Contact Info

Product

Resources

About