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Dissecting the pathways of doom - application of spectroscopy in the study of cell death and survival

100%

Godlewski M. , Gorka M. , Gajkowska B. , Wojewodzka U.

tom Vol. 36, nr 2-3

257-270

Biological studies rely on measurements performed on large populations, which allow the statistic evaluation of gathered data. On the other hand, the need to precisely localize observed processes and the facts occurring in the microscale results in the need of high resolution qualitative imaging. These two approaches are complementary and without anyone of them no valuable experimental data can be gathered from studies of fluorescent dyes. Both approaches are based on common origin of laser-induced fluorescence, but due to the method of analysis they split into the quantitative (laser scanning cytometry) and qualitative (confocal microscopy) fields of research.

75%

Zarzynska J. , Motyl T.

nr 3

181-193

In mammary epithelial cells (MEC) TGF-ß1 is the auto-/paracrine growth inhibitor and inducer of apoptosis and therefore is considered as an important local regulator of mammary tissue involution. However, the mechanisms of controlled TGF-ß1 expression in the course of bovine mammary gland remodelling are still unclear. Recent study performed in this laboratory support the evidence that TGF-ß1 expression in bovine MEC is regulated by hormones of somatotropic axis (GH, IGF-I and somatostatin). Present study was focused on the contribution of IGF-I-induced signaling pathways in anti-TGF-ß1 and anti-apoptotic effects of IGF-I. Laser scanning cytometry was applied for the measurement of TGF-ß1 content and apoptotic cell number in bovine BME-UV1 MEC. Involution of the bovine mammary gland in vitro was modeled by decreasing the availability of FBS for bovine MEC. Reducing FBS content in the medium from 10% to 0.5% evoked highly significant increase of TGF-ß1 expression and increase of apoptotic cell number. IGF-I (50 ng/ml) completely abrogated FBS deficiency-induced TGF-ß1 expression and apoptosis in bovine MEC. In order to establish which of the IGF-I signaling pathways contributed to anti-TGF-ß1 and anti-apoptotic effects, the inhibitors of PI3-kinase - (LY 294002) and MEK- (MAPKK for ERK) (PD 098059) mediated signaling pathways were applied to our model. The results clearly showed that inhibition of PI3-K reverses the ability of IGF-I to suppress TGF-ß1 expression and apoptosis. An inhibition of ERK1/2 pathway even potentiated inhibitory effect of IGF-I on TGF-ß1 expression, but partially abrogated anti-apoptotic effect of IGF-I. In conclusion, the results of the study indicate that PI3-K/Akt pathway contributed significantly to the inhibition of TGF-ß1 expression by IGF-I, whereas both PI3-K/Akt and ERK1/2 pathways are involved in the anti-apoptotic effect of IGF-I in bovine MEC.