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Besides neuro-imaging, the Thy1-YFP mouse could serve for visualizing experimental tumours, inflammation and wound-healing.

Jósvay K, Winter Z, Katona RL, Pecze L, Marton A, Buhala A, Szakonyi G, Oláh Z, Vizler C - Sci Rep (2014)

Bottom Line: The Thy1 (CD90) cell surface glycoprotein is present on many cell types besides neurons, and is known to be involved in cell adhesion, migration and signal transduction.We hypothesized that Thy1-activating conditions could probably activate the truncated Thy1 regulatory sequences used in the Thy1-YFP construct, resulting in YFP transgene expression outside the nervous system.In the tumour mass, the yellow-green fluorescent stromal cells were clearly distinguishable from 4T1 carcinoma cells stably transfected with red fluorescent protein.

View Article: PubMed Central - PubMed

Affiliation: 1] Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary [2] Institute of Pharmaceutical Analysis, Faculty of Pharmacy, University of Szeged, Szeged, Hungary.

ABSTRACT
The B6.Cg-Tg(Thy1-YFP)16Jrs/J transgenic mouse strain, widely used to study neuronal development and regeneration, expresses the yellow fluorescent protein (YFP) in the peripheral nerves and the central nervous system under the control of regulatory sequences of the Thy1 gene. The Thy1 (CD90) cell surface glycoprotein is present on many cell types besides neurons, and is known to be involved in cell adhesion, migration and signal transduction. We hypothesized that Thy1-activating conditions could probably activate the truncated Thy1 regulatory sequences used in the Thy1-YFP construct, resulting in YFP transgene expression outside the nervous system. We demonstrated that the stroma of subcutaneous tumours induced by the injection of 4T1 or MC26 carcinoma cells into BALB/c(Thy1-YFP) mice, carrying the same construct, indeed expressed the YFP transgene. In the tumour mass, the yellow-green fluorescent stromal cells were clearly distinguishable from 4T1 carcinoma cells stably transfected with red fluorescent protein. Local inflammation induced by subcutaneous injection of complete Freund's adjuvant, as well as the experimental wound-healing milieu, also triggered YFP fluorescence in both the BALB/c(Thy1-YFP) and B6.Cg-Tg(Thy1-YFP)16Jrs/J mice, pointing to eventual overlapping pathways of wound-healing, inflammation and tumour growth.

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Wound healing activates the Thy-1 promoter in Thy1-YFP mice.The ear pinnae of BALB/c(Thy-1-YFP) mice were cut with a scalpel under aseptic conditions, under anaesthesia, and the wound healing was then followed by fluorescent microscopy. The wound-healing process was accompanied by temporary upregulation of the YFP fluorescence, gradually fading after 2 weeks. Photographs of the mouse ears were made under light anaesthesia, using a Nikon Eclipse E600 fluorescent microscope (either 20× magnification objective lens or 10× magnification objective lens). Some of the pictures were generated by the combination of multiple images to depict the whole wound surface.
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f4: Wound healing activates the Thy-1 promoter in Thy1-YFP mice.The ear pinnae of BALB/c(Thy-1-YFP) mice were cut with a scalpel under aseptic conditions, under anaesthesia, and the wound healing was then followed by fluorescent microscopy. The wound-healing process was accompanied by temporary upregulation of the YFP fluorescence, gradually fading after 2 weeks. Photographs of the mouse ears were made under light anaesthesia, using a Nikon Eclipse E600 fluorescent microscope (either 20× magnification objective lens or 10× magnification objective lens). Some of the pictures were generated by the combination of multiple images to depict the whole wound surface.

Mentions: In response to tissue injury, and during the subsequent wound-healing process, sequential and overlapping events take place. First, the active bleeding must be stopped, then inflammatory, proliferative and tissue remodelling phases occur, which include the activation and migration of distinct sets of immune cells, stem cells, fibroblasts and keratinocytes. Since Thy1 is known to affect cell migration into the wound area, we followed the changes in YFP fluorescence during the healing process. One day after sharp superficial cuts were made in the ear of either B6.Cg-Tg(Thy1-YFP)16Jrs/J or BALB/c(Thy1-YFP) mice by scalpel in anesthesia, a faint fluorescent halo appeared along the wound, which expanded until day 14. At the end of day 3, a more intense fluorescent margin also emerged around the wound, which remained significantly brighter until the end of the experiment. The fluorescence area around the wound gradually decreased from day 21, and had almost entirely disappeared by the end of day 36 (Figure 4). As in the case of the local inflammation model, the fluorescence intensity of the YFP transgene was much above the autofluorescence of non-transgenic mice receiving the same treatment (Supplementary Figure 2).


Besides neuro-imaging, the Thy1-YFP mouse could serve for visualizing experimental tumours, inflammation and wound-healing.

Jósvay K, Winter Z, Katona RL, Pecze L, Marton A, Buhala A, Szakonyi G, Oláh Z, Vizler C - Sci Rep (2014)

Wound healing activates the Thy-1 promoter in Thy1-YFP mice.The ear pinnae of BALB/c(Thy-1-YFP) mice were cut with a scalpel under aseptic conditions, under anaesthesia, and the wound healing was then followed by fluorescent microscopy. The wound-healing process was accompanied by temporary upregulation of the YFP fluorescence, gradually fading after 2 weeks. Photographs of the mouse ears were made under light anaesthesia, using a Nikon Eclipse E600 fluorescent microscope (either 20× magnification objective lens or 10× magnification objective lens). Some of the pictures were generated by the combination of multiple images to depict the whole wound surface.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4209462&req=5

f4: Wound healing activates the Thy-1 promoter in Thy1-YFP mice.The ear pinnae of BALB/c(Thy-1-YFP) mice were cut with a scalpel under aseptic conditions, under anaesthesia, and the wound healing was then followed by fluorescent microscopy. The wound-healing process was accompanied by temporary upregulation of the YFP fluorescence, gradually fading after 2 weeks. Photographs of the mouse ears were made under light anaesthesia, using a Nikon Eclipse E600 fluorescent microscope (either 20× magnification objective lens or 10× magnification objective lens). Some of the pictures were generated by the combination of multiple images to depict the whole wound surface.
Mentions: In response to tissue injury, and during the subsequent wound-healing process, sequential and overlapping events take place. First, the active bleeding must be stopped, then inflammatory, proliferative and tissue remodelling phases occur, which include the activation and migration of distinct sets of immune cells, stem cells, fibroblasts and keratinocytes. Since Thy1 is known to affect cell migration into the wound area, we followed the changes in YFP fluorescence during the healing process. One day after sharp superficial cuts were made in the ear of either B6.Cg-Tg(Thy1-YFP)16Jrs/J or BALB/c(Thy1-YFP) mice by scalpel in anesthesia, a faint fluorescent halo appeared along the wound, which expanded until day 14. At the end of day 3, a more intense fluorescent margin also emerged around the wound, which remained significantly brighter until the end of the experiment. The fluorescence area around the wound gradually decreased from day 21, and had almost entirely disappeared by the end of day 36 (Figure 4). As in the case of the local inflammation model, the fluorescence intensity of the YFP transgene was much above the autofluorescence of non-transgenic mice receiving the same treatment (Supplementary Figure 2).

Bottom Line: The Thy1 (CD90) cell surface glycoprotein is present on many cell types besides neurons, and is known to be involved in cell adhesion, migration and signal transduction.We hypothesized that Thy1-activating conditions could probably activate the truncated Thy1 regulatory sequences used in the Thy1-YFP construct, resulting in YFP transgene expression outside the nervous system.In the tumour mass, the yellow-green fluorescent stromal cells were clearly distinguishable from 4T1 carcinoma cells stably transfected with red fluorescent protein.

View Article: PubMed Central - PubMed

Affiliation: 1] Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary [2] Institute of Pharmaceutical Analysis, Faculty of Pharmacy, University of Szeged, Szeged, Hungary.

ABSTRACT
The B6.Cg-Tg(Thy1-YFP)16Jrs/J transgenic mouse strain, widely used to study neuronal development and regeneration, expresses the yellow fluorescent protein (YFP) in the peripheral nerves and the central nervous system under the control of regulatory sequences of the Thy1 gene. The Thy1 (CD90) cell surface glycoprotein is present on many cell types besides neurons, and is known to be involved in cell adhesion, migration and signal transduction. We hypothesized that Thy1-activating conditions could probably activate the truncated Thy1 regulatory sequences used in the Thy1-YFP construct, resulting in YFP transgene expression outside the nervous system. We demonstrated that the stroma of subcutaneous tumours induced by the injection of 4T1 or MC26 carcinoma cells into BALB/c(Thy1-YFP) mice, carrying the same construct, indeed expressed the YFP transgene. In the tumour mass, the yellow-green fluorescent stromal cells were clearly distinguishable from 4T1 carcinoma cells stably transfected with red fluorescent protein. Local inflammation induced by subcutaneous injection of complete Freund's adjuvant, as well as the experimental wound-healing milieu, also triggered YFP fluorescence in both the BALB/c(Thy1-YFP) and B6.Cg-Tg(Thy1-YFP)16Jrs/J mice, pointing to eventual overlapping pathways of wound-healing, inflammation and tumour growth.

Show MeSH
Related in: MedlinePlus