Detection of T-Cadherin Expression in Mouse Embryos

Abstract

The aim of the present study was to evaluate T-cadherin expression at the early developmental stages of the mouse embryo. Using in situ hybridization and immunofluorescent staining of whole embryos in combination with confocal microscopy, we found that T-cadherin expression is detected in the developing brain, starting with the E8.75 stage, and in the heart, starting with the E11.5 stage. These data suggest a possible involvement of T-cadherin in the formation of blood vessels during embryogenesis.

Keywords: T-cadherin; angiogenesis; embryogenesis; in situ hybridization.

Fig. 1

In situ hybridization of mouse embryos at the of E8.75 stage. Expression of T-cadherin mRNA (T-cad): 1 – in the mesencephalon region; 2 – in the base of the developing optic vesicle; 3 – in the inner lining of the telencephalon; 4 – in the myelencephalon; 5, 6 – in the optic vesicles. No specific staining in the negative control (control). Magnification of 3.2, 5, and 6×

Fig. 2

In situ hybridization of mouse embryos at the E9.5 stage. Staining of brain regions corresponds to the localization of T-cadherin mRNA (T-cad). Expression is observed in the base of the developing optic vesicles, in the parietal and occipital bend regions. Small arrows indicate the base of the developing optic vesicle. Control – the negative control. Magnification of 3.2, 5, and 6×

Fig. 3

In situ hybridization of mouse embryos at the E10.5 stage. Intense expression of T-cadherin (T-cad) in the developing tectum and the lateral regions of the diencephalon. The arrow indicates specific staining of the choroid plexus in the telencephalon region. No specific staining in the negative control (control). Magnification of 3.2, 5, and 6×

Fig. 4

In situ hybridization of mouse embryos at the E10.5 stage. T-cad – specific staining of T-cadherin in the tectum (in the occipital bend region) and inner lining of the telencephalon; control – no specific staining in the negative control; Krox20 – staining of central nervous system structures in the positive control. Magnification of 3.2×

Fig. 5

Immunofluorescent staining of mouse embryos at the E9.5 (-1-) and E12.5 (-2-) stages. Specific staining (red fluorescence) corresponds to T-cadherin expression in the linings of the brain at both stages; expression of T-cadherin in the developing optic vesicle in the E9.5 embryo. Blue fluorescence corresponds to nuclei additionally stained with DAPI. Magnification of 5×

Fig. 6

Immunofluorescent staining of mouse embryos at the E11.5 stage. Specific staining (red fluorescence) corresponds to expression of the T-cadherin protein. Blue fluorescence corresponds to nuclei additionally stained with DAPI. 1 – specific staining in the diencephalon region, as well as in the region of the developing eyecup; 2 – specific staining in the mesencephalon and metencephalon region; 3 – the same region as in 2 – at another optical plane level. Magnification of 5×

Fig. 7

Immunofluorescent staining of mouse embryos at the E11.5 stage. Specific staining (red fluorescence) corresponds to expression of the T-cadherin protein (T-cad). Blue fluorescence corresponds to nuclei additionally stained with DAPI. 1 – specific staining reflecting T-cadherin expression in the heart region; 2 – control staining with antibodies to immunoglobulin G. Magnification of 5×

Fig. 8

Lack of T-cadherin mRNA in the developing heart of mouse embryos at the E8.75–E10.5 stages (1, 2, 3). Arrows and the selected area indicate the developing heart region. Magnification of 5× (1, 2) and 6× (3)