engleski

Immunohistochemical expression of HNE-protein conjugates in rat experimental teratoma

4-hydroxynonenal (HNE) is a marker of oxidative stress and also seems to influence cell proliferation and differentiation. To investigate the distribution of HNE-protein conjugates in various embryonal tissues, we have used rat teratoma obtained by specific method of in vitro cultivation of the rat embryo. On 9, 5th day of pregnancy, embryos were isolated from the uterus of pregnant female rat. Extraembryonal part was discarded and only the embryo-proper containing three germ-layers was cultivated on a lens paper at the air-liquid interface supported by a steel grid for 14 days. Eagle's MEM was supplemented with 50% rat serum. After 14 days, teratomas developed and were fixed in St. Marie's fixative and processed for immunohistochemistry. Monoclonal antibody against 4-hydroxynonenalhistidine conjugates (Waeg et al, 1996) (HNE) was applied and detected by Dako LSAB Kit (Dako, K0609). HNE was found to be widely distributed in cytoplasm of cells in tissue derivates of all three germ-layers. Strong signal was present in epidermis and neuroblasts. Myoblasts and myotubes have shown a slightly weaker signal and in connective tissue the expression was seemingly related to the density of extracellular matrix. Gut-like tissue and respiratory-like epithelium were both strongly positive and only the cartilage was negative for HNE. The distribution of HNE in experimental rat teratoma seems to be tissue-specific and therefore possibly related to its role in cell proliferation and differentiation.

4-hydroxynonenal; HNE; oxidative stress; HNE-protein conjugates; rat embryo; rat teratoma; myoblasts; myotubes; neuroblasts

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