NFAT2 regulates COX-2 expression and modulates the integrin repertoire in endothelial cells at the crossroads of angiogenesis and inflammation.

The mechanisms controlling the switch between the pro-angiogenic and pro-inflammatory states of endothelial cells are still poorly understood. In this paper, we show that: (a) COX-2 expression induced by VEGF-A is NFAT2-dependent; and (b) the integrin profile in endothelial cells induced by the pro-angiogenic VEGF-A is distinct from that brought on by the inflammatory cytokine TNF-α. Two groups of integrin subunits specifically upregulated over time by both cytokines were identified using RT-PCR and Western Immunoblotting.

Bacterial expression, purification and angiogenesis-promoting activity of human thymosin β4.

Thymosin β4 (Tβ4) is an actin-binding peptide involved in tissue regeneration and angiogenesis. This 43-amino acid peptide is chemically synthesized for research or clinical trials. To overcome the high costs of solid phase synthesis, we developed a genetic engineering procedure of Tβ4 expression in a protease-deficient host strain, Escherichia coli BL21(DE3), transformed with different expression vectors (pRSETA, pET-15b and pEcoli-Cterm6 × HN).

Gene expression pattern in PC12 cells with reduced PMCA2 or PMCA3 isoform: selective up-regulation of calmodulin and neuromodulin.

Cellular calcium homeostasis is controlled predominantly by the plasma membrane calcium pump (PMCA). From four PMCA isoforms, PMCA1 and PMCA4 are ubiquitous, while PMCA2 and PMCA3 are found in excitable cells. We have previously shown that suppression of neuron-specific PMCAs in non-differentiated PC12 cells changed the cell morphology and triggered neuritogenesis.

Adaptation of microsomal glutathione transferase 1 in PC12 cells with modified PMCA isoforms composition.

Microsomal glutathione transferase 1 (MGST1) is an integral homo-trimeric membrane protein with transferase and peroxidase activities. With glutathione as a co-substrate, it scavenges toxic compounds and may exert anti-apoptotic effect. We examined the effect of suppression of plasma membrane Ca(2+)-ATPase isoforms--PMCA2 or PMCA3 on MGST1 in PC12 cells.

Functional characteristic of PC12 cells with reduced microsomal glutathione transferase 1.

Microsomal glutathione transferase 1 (MGST1) possesses glutathione transferase and peroxidase activities and is active in biotransformation of xenobiotics and in defense against oxidative stress. To assess MGST1 role in the development and functioning of PC12 cells, we constructed a cell line with reduced MGST1 (PC12_M). Real-time PCR and immunoblot assays showed MGST1 expression lowered to 60 % and immunocytochemical analyses demonstrated an altered concentration and distribution of the enzyme.

Fast action of neuroactive steroids on plasma membrane calcium pump in PC12 cells.

Calcium ions are essential to proper neurotransmission. Impairment in cytosolic Ca(2+) concentration and Ca(2+) signaling disturbs neuronal activity, leading to pathological consequences. In cells, a high-affinity plasma membrane calcium pump (PMCA) keeps free Ca(2+) in the nanomolar range.

Changes in erythrocyte glutathione and plasma membrane calcium pump in preterm newborns treated antenatally with MgSO4.

Background: Magnesium is required for the proper activity of many metabolic pathways in every cell type. Mg deficiency gives rise to preterm birth and low body weight that are associated with pathological circumstances, including disturbed ions homeostasis and insufficient antioxidant protection. Antenatal MgSO(4) treatment has been reported to exhibit a protective effect on the developing fetus; however, the molecular mechanism of this protection remains not fully understood.

Functional importance of PMCA isoforms in growth and development of PC12 cells.

Intracellular Ca2+ in neuronal cells is an essential regulatory ion responsible for excitability, synaptic plasticity, and neurite outgrowth. Plasma membrane calcium ATPase (PMCA) is the most sensitive enzyme in decreasing of the Ca2+ concentration. The diverse PMCA isoforms composition in the membranes suggests their specific function in the cell, and whereas PMCA1 and 4 appear to be ubiquitous, PMCA2 and 3 are characteristic isoforms for excitable cells.