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Annexins as mediators in intracellular signalling pathways - are they intracellular GTP biosensors?

100%

Pikula S. , Danieluk M. , Golczak M. , Kirilenko A. , Bandorowicz-Pikula J.

nr Suppl.

PP2A controls methionine metabolism elicited by intracellular H2O2

84%

Trotta A. , Li S. , Mhamdi A. , Ravanel S. , Noctor G. , Kangasjarvi S.

nr 2

Mobilization of intracellular calcium by intracellular flash photolysis of caged dihydrosphingosine in cultured neonatal rat sensory neurones

84%

Ayar A. , Thatcher N. M. , Zehavi U. , Trentham D. R. , Scott R. H.

nr 2

The ability of dihydrosphingosine to release Ca2+ from intracellular stores in neurones was investigated by combining the whole cell patch clamp technique with intracellular flash photolysis of caged, N-(2-nitrobenzyl)dihydrosphingosine. The caged dihydrosphingosine (100 μM) was applied to the intracellular environment via the CsCl-based patch pipette solution which also contained 0.3% dimethylformamide and 2 μM dithiothreitol. Cultured dorsal root ganglion neurones from neonatal rats were voltage clamped at -90 mV and inward whole cell Ca2+-activated currents were recorded in response to intracellular photorelease of dihydrosphingosine. Intracellular photorelease of dihydrosphingosine (about 5 μM) was achieved using a Xenon flash lamp. Inward Ca2+-activated currents were evoked in 50 out of 57 neurones, the mean delay to current activation following photolysis was 82±13 s. The responses were variable with neurones showing transient, oscillating or sustained inward currents. High voltage-activated Ca2+ currents evoked by 100 ms voltage step commands to 0 mV were not attenuated by photorelease of dihydrosphingosine. Controls showed that alone a flash from the Xenon lamp did not activate currents, and that the unphotolysed caged dihydrosphingosine, and intracellular photolysis of 2-(2-nitrobenzylamino) propanediol also did not evoke responses. The dihydrosphingosine current had a reversal potential of -11±3 mV (n = 11), and was carried by two distinct Cl- and cation currents which were reduced by 85% and about 20% following replacement of monovalent cations with N-methyl-D-glucamine or application of the Cl- channel blocker niflumic acid (10 μM) respectively. The responses to photoreleased dihydrosphingosine were inhibited by intracellular application of 20 μM EGTA, 10 μM ryanodine or extracellular application of 10 μM dantrolene, but persisted when Ca2+ free saline was applied to the extracellular environment. Intracellular application of uncaged dihydrosphingosine evoked responses which were attenuated by photolysis of the caged Ca2+ chelator Diazo-2. Experiments also suggested that extracellular application of dihydrosphingosine can activate membrane conductances. We conclude that dihydrosphingosine directly or indirectly mobilises Ca2+ from ryanodine-sensitive intracellular stores in cultured sensory neurones.

Abnormal intracellular signalling in skin fibroblasts from LNS patients and HGPRT-deficient neuroblastoma

84%

Hussain S. P. , Alexander N. , Duley J. A. , Connolly G. P.

nr 3

Study of cell survival in non-adherent conditions

84%

Malecki M. , Sroczynska P. , Janik P.

tom 05

nr 2

Signalling via reactive oxygen species – why and how?

67%

Bartosz G.

nr 2

Effects of dietary fish oil on NF kappa B gene expression and related signaling in spleen of chickens stimulated with lipopolysaccharide

67%

Yang X. , He X. , Zhang B. , Yang Y. , Yuan J. , Guo Y.

nr 2

The study was conducted to investigate the effects of fish oil and maize oil on nuclear factor kappa B (NFκB) gene expression and the downstream pathways of intracellular signaling in spleen of chickens after lipopolysaccharide (LPS) stimulation. Two hundred eighty eight chickens were assigned in a 2×2 factorial design. Factors were dietary fat type (4.5% maize oil or 4.5% fish oil) and immunological stimulation (LPS or saline). LPS increased levels of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and arachidonic acid (AA) of spleen in chickens after the second LPS stimulation on 28 d of age. Fish oil alleviated the increase of EPA and DHA in spleen of chickens after LPS stimulation at 27 d of age. Fish oil supplementation decreased prostaglandin 2 (PGE2) production and the activity of cyclooxygenase 2 (COX2) after LPS stimulation. LPS stimulation increased the activity of phospholipase C (PLC) in spleen of chickens. And fish oil inhibited activity of PLC in spleen of chickens stimulated by LPS. Meanwhile fish oil decreased the production of inositol triphosphate (IP3) in spleen of chickens stimulated by LPS. Fish oil alleviated the mRNA abundance elevation of nuclear factor kappa B (NFκB) after LPS stimulation. These results showed that fish oil down-regulated the production of IP3 and PGE2 through inhibiting the activity of PLC and COX2 in spleen of chickens, respectively. The results of NFκB gene expression suggested fish oil might alleviate immune stress at the level of transcription.