SVIBOR - Project code: 3-01-006


Strossmayerov trg 4, HR - 10000 ZAGREB
tel.: +385 1 459 44 44, fax: +385 1 459 44 69


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Project code: 3-01-006

Second messengers and mechanisms of ion transport in the cells

Main researcher: BANFIĆ, HRVOJE (96771)

Type of research: basic
Duration from: 03/01/91. to 02/28/94.

Papers on project (total): 0
Institution name: Medicinski fakultet, Zagreb (108)
Department/Institute: Departement for physiology
Address: Šalata 3, Zagreb
Phone: 385 (0)466-920
Fax: 385 (0)424-001

Summary: In proposed project second messengers related to inositol lipid metabolism will be studied in isolated plasma membranes obtained from rat renal cortex upon the stimulation of cortical slice with different hormons or growth factors. Inositol lipid metabolism will also be studied in cell nuclei during compensatory renal and hepatic growth.Transport mechanisms for different ions (sodium, hydrogen, bicarbonate and calcium) will be studied in isolated cell membranes and whole cells. It will be also investigated mechanisms of activation of phospholipase A2 and protein kinase C isoformes by phorbol-esters and interferon gamma.

Keywords: second messengers, cell membrane, cell nuclei, ion transport, endothelins, phorbol esters, protein kinase C, phospholipase A2, interferon gamma

Research goals: 1. Development of technics for direct determination secondmessengers mass and their outrunners (1,2-diacil-glicerol,inositol 1,4,5-triphosphate, phosphatidil-inositol,phosphatidil-inositol 4-monophosphate and phosphatidil-inositol4,5-biphosphate). 2. Determination hormon-dependent and hormon-independentinositol lipid concentration, as well as measurement of secondmessenger production by action of different hormons and growthfactors in brush border and basal lateral membranes prepared fromrat kidney cortex. 3. Measurement of second messengers and arahidonic acidmetabolism in permanent cell cultures by action of differentlimfokines. 4. Measurement of inositol lipid concentration and secondmessengers in cell nuclei isolated from rat kidney and liverduring process of compensatory growth. 5. Determination of Na and H exchange activity, conductivity forH and K, as well as NEM-dependent ATP-ase activity in brushborder membrnes isolated from rat kidey cortex duringcompensatory kidney growth. 6. Demonstration of Na and HCO cotransportin basal lateralmembranes prepared frok rat kidney cortex, activity of potentialinhibitors (amilorid, bumetanid and etacrinic acid) and effectsof unilateral nephrectomy on cotransport activity. 7. Determination of protein kinase C activity, phosphorilation ofproteins in cell membranes during compensatory kidney growth andactions of different hormons. 8. Demonstrstion of phosphatidil-cholin specific phospholipase Cin cell membranes isolated from rat kidney cortex. 9. Isoaltion of renotropic factor which start compensatory kidneygrowth. 10. Investigation of gluconeogenetic ability of kidney cortex innormal and uninephectomized rats under high-protein andhigh-carbohydrate diet, as well as effect of gluco-corticoids ongluconeogenesis in this dietary conditions. 11. Determination of free intracellular calcium concentration andchange of intracellular pH in kidney epitelialcells by actions ofhormons and growth factors, as well as during compensatory kidneygrowth. 12. Effects of acidosis and alcalosis, diabetes, adrenalectomy,gluco-corticoid treated adrenalectomised animals andinsulin treated diabetic animals on Na and H exchange, conductionfor cations, activity of Na/K ATP-ase and H-ATPase in homogenate,luminal and contraluminal membranes isolated from proksimaltubuls of rat kidney. 13. Studing of transporter for sodium and hydrogen, calciumATP-ase and hydrogen pump in eritrocite "inside-out" and"right-side-out" membrane vesicles prepared from rat blood. 14. Studing of trnsport process activation with secondmessengers. 15. Studing the signalling pathway which involves phospholipase A2 activation Expected result will be finding out basic physiologicalmechanisms of production of second messengers which mobilizecalcium in cell membranes and cell nucleus, by action ofdifferent hormons and growth factors. Furthermore, importantwill be finding of transport process mechanisms for each ionin isolated cell membranes and cells, as well as cell metabolismby action of hormons, second messengers and differentphysiological factors.

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Last update: 10/09/95