engleski

Ganglioside expression in tissues of mice lacking the tumor necrosis factor receptor 1

The aim of this study was to investigate ganglioside expression in lymphoid (spleen and thymus) and other tissues (lungs, brain, muscle, liver) of the mice lacking tumor necrosis factor receptor 1(TNFRp55). Gangliosides were isolated from C57BL/6 mice homozygous (-/-) or heterozygous (-/+) for the TNFR gene knockout and separated by high performance thin layer chromatography (HPTLC). Individual gangliosides were visualized using resorcinol stain and HPTLC immunostaining overlay technique with polyclonal antibodies specific for GM3(Neu5Ac), GM3 (Neu5Gc), GalNAcGm1b and nLc4. GM1b-type gangliosides were detected after V. cholerae neuraminidase treatment with a specific anti-Gg4 antibody. Neuraminidase pretreatment was also used to detect gangliosides IV3nLc4 and VI3nLc6. Individual HPTLC bands were scanned with a CD60 scanner to quantify the differnce between -/- and -/+ mice. Immunostaining showed that GM3(Neu5Ac) was ten times and GM3(Neu5Gc) seven toimes more abundant oin the lungs of -/+ mice than of -/- mice. Two bands of GM1b were abundantly expressed in the lungs, spleen and thymus of -/+ mice, but not in the lungs of the -/- mice, whereas the amounts of this ganglioside in the spleen and thymus of -/- mice were at least ten times lower in -/+ mice . Two bands of GalNAcGM1b were expressed in the lungs, spleen and thymus of -/+ mice but not in the lungs anfd thymus of -/- mice; only traces of the lower GalNAc-GM1b band were observed in the spleen of -/- mice. Gangliosides of the neolacto-series were several times more abundant in the lungs, brain and muscle of -/- mice then of -/+ mice. Our study provides in vivo evidence that TNF signaling via TNFRp55 is involved in the acquisition of distinct ganglioside profiles of different mouse organs. TNFRp55 signaling seems to be especially important for the activation of the GM1b ganglioside biosynthetic pathway which is a unique characteristic of mouse immune tissues.

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