Glycosyl phosphatidylinositol (GPI)/inositolphosphate glycan (IPG): an intracellular signalling system involved in the control of thyroid cell proliferation.

In porcine thyrocytes, TSH alone does not induce cell growth. Recently, it has been demonstrated that acute stimulation by TSH of porcine thyrocytes leads to release an inositolphosphate glycan (IPG) described as a putative second messenger for various growth factors in different cell types. IPG isolated from porcine thyrocytes induces proliferation of fibroblasts EGFR T17 and porcine thyrocytes. In porcine thyrocytes we have confirmed that cell growth requires the presence of both TSH and insulin. This effect is reproduced by 8-bromo cyclic AMP suggesting a mediation by intracellular cyclic AMP. Cooperative effects between 8-bromo cyclic AMP and IPG have also been evidenced and are in favour of a crosstalk between distinct signalling pathways.

Cyclic AMP regulation of annexins I, II, V synthesis and localization in cultured porcine thyroid cells.

Porcine thyroid cells cultured in the presence of TSH (0.1 mU/ml) or forskolin (10(-5) M) for 4 days display an increased annexin I, II, V biosynthesis when compared with unstimulated cells. Annexin I mostly accumulates in the cytosolic fraction and annexins II and V in the particulate fraction. TSH and forskolin affect in the same manner annexin biosynthesis and localization. In the TSH and forskolin treated cells PGE2 production is very low in comparison with the very high PLA2 activity observed in the dedifferentiated control cells. A strong inhibition of the PGE2 production is observed in control cells incubated with a purified annexin V preparation. These results suggest the existence, in porcine thyroid cells, of a cross-talk between the adenylyl cyclase system and the phospholipase A2 mediated pathways. Annexins' biosynthesis and localization are under the control of the adenylyl cyclase system and participate in this cross-talk.

Production of lipocortin-like proteins by cultured human tracheal submucosal gland cells.

Evidence is obtained for the presence of lipocortin-like proteins in human tracheal gland cells in culture. Using polyclonal antibodies to lipocortin I, indirect immunofluorescence studies demonstrate that lipocortin I is mainly confined to the tracheal gland cell surface. From cell membranes, four Ca2(+)-dependent proteins (35, 40, 45 and 67 kDa) were identified as lipocortin related proteins by using immunoblotting and fluorography following [35S]methionine metabolic labeling experiments. A strong immunoreactivity for the 35 kDa protein was observed. In addition, lipocortin-like proteins with apparent Mr33, 35, 37 and 67 kDa, respectively, were released in the apical culture medium by tracheal gland cells cultured on microporous membrane of a double chamber culture system.