Second messengers and mechanisms of ion transport in the cells
Main researcher
: BANFIĆ, HRVOJE (96771) Assistants
ANDREIS, ANA (505)
CRNOGORAC-JURČEVIĆ, TATJANA (80782)
DEMBIĆ, ZLATKO (81634)
GABRIĆ, DORA (12751)
GATALICA, ZORAN (138016)
HADŽIĆ, AMELA (160755)
KUKOLJA, SUNČANA (112980)
MAČKOVIĆ-BAŠIĆ, MARIJA (54783)
POKRAJAC, NIKŠA (37826)
RIES, NIKOLA (41016)
SALIHAGIĆ, AIDA (140090)
SOLJAČIĆ, MARIJA (44082)
ZIMOLO, ZORAN (160766)
BEARA-LASIĆ, LADA (900071)
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
Communication
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. Other information about the project.