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GFP Mice (gfp + mouse)

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Medical Sciences	100%

Selected Abstracts

In vivo application of mAb directed against the ,, TCR does not deplete but generates "invisible" ,, T cells

EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 2 2009
Christian Koenecke
Abstract mAb targeting the ,, TCR have been used for ,, T-cell depletion with varying success. Although the depletion-capacity of the anti-,, TCR mAb clone GL3 has been disputed repeatedly, many groups continue to use ,, T-cell depletion protocols involving the mAb clone UC7-13D5 and find significant biological effects. We show here that treatment with both GL3 and UC7-13D5 antibodies does not deplete ,, T cells in vivo, but rather leads to TCR internalization and thereby generates "invisible" ,, T cells. We addressed this issue using anti-,, TCR mAb injections into WT mice as well as into reporter TCR delta locus-histone 2B enhanced GFP knock-in mice, in which ,, T cells can be detected based on an intrinsic green fluorescence. Importantly, the use of TCR delta locus-histone 2B enhanced GFP mice provided here for the first time direct evidence that the "depleted" ,, T cells were actually still present. Our results show further that GL3 and UC7-13D5 mAb are in part cross-competing for the same epitope. Assessed by activation markers, we observed in vitro and in vivo activation of ,, T cells through mAb. We conclude that ,, T-cell depletion experiments must be evaluated with caution and discuss the implications for future studies on the physiological functions of ,, T cells. [source]

Transplanted XY germ cells produce spermatozoa in testes of XXY mice,

INTERNATIONAL JOURNAL OF ANDROLOGY, Issue 4 2010
Y. Lue
Summary XXY mouse has been characterized as an experimental model for men with Klinefelter's syndrome (XXY male phenotype). To test whether donor XY germ cells could proliferate and differentiate in the XXY testicular environment, donor testicular cells from adult (2,3 months old) and immature (10 days old) XY green fluorescence protein (GFP) transgenic mice were transplanted into the seminiferous tubules of adult (4,7 months old) and young (6 weeks old) XXY recipient mice respectively. Twelve weeks after transplantation, GFP positive spermatogonia were found in 21.74% (five out of 23) of adult XXY recipients who received adult donor cells. The GFP positive segments of seminiferous tubules were observed in 44.44% (four out of nine) young XXY recipients who received donor cells from 10 days old GFP mice. We found using immunohistochemistry and cell morphology that donor-derived GFP positive germ cells were spermatogonia, spermatocytes, round spermatids and spermatozoa in some of the seminiferous tubules of young XXY recipient mice. The results demonstrated that the donor XY germ cells were able to qualitatively complete spermatogenesis in some of the seminiferous tubules of XXY mice. [source]

Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging

MICROSCOPY RESEARCH AND TECHNIQUE, Issue 12 2006
Wen Lo
Abstract The aim of this work is to demonstrate that multiphoton microscopy is a preferred technique to investigate intact cornea structure without slicing and staining. At the micron resolution, multiphoton imaging can provide both large morphological features and detailed structure of epithelium, corneal collagen fibril bundles and keratocytes. A large area multiphoton cross-section across an intact eye excised from a GFP mouse was obtained by a homebuilt multiphoton microscope. The broadband multiphoton fluorescence (435,700 nm) and second harmonic generation (SHG, 360,400 nm) signals were generated by the 760 nm output of a femtosecond titanium-sapphire laser. A water immersion objective (Fluor Ô, 40X, NA 0.8; Nikon) was used to facilitate imaging the curve ocular surface. The multiphoton image over entire cornea provides morphological information of epithelial cells, keratocytes, and global collagen orientation. Specifically, our planar, large area multiphoton image reveals a concentric pattern of the stroma collagen, indicative of the laminar collagen organization throughout the stroma. In addition, the green fluorescence protein (GFP) labeling contributed to fluorescence contrast of cellular area and facilitated visualizing of inactive keratocytes. Our results show that multiphoton imaging of GFP labeled mouse cornea manifests both morphological significance and structural details. The second harmonic generation imaging reveals the collagen orientation, while the multiphoton fluorescence imaging indicates morphology and distribution of cells in cornea. Our results support that multiphoton microscopy is an appropriate technology for further in vivo investigation and diagnosis of cornea. Microsc. Res. Tech., 2006. © 2006 Wiley-Liss, Inc. [source]