In Chlamydomonas eugametos gametes, phosphatidylinositol 4-phosphate (PtdInsP) and phosphatidylinositol 4,5-bisphosphate (PtdInsP2) comprised 0.4 and 0.3% of the whole-cell phospholipids. They were concentrated in the plasma membrane around the cell body and were present in low concentrations in the flagellar membrane. When gametes were fed (32)PO 4 (-) , the label was rapidly incorporated into PtdInsP and PtdInsP2 and only slowly incorporated into structural lipids such as phosphatidylethanolamine and phosphatidylglycerol. Similarly, when a pulse of (32)PO 4 (-) was chased with PO 4 (-) , the label was rapidly lost from the polyphosphoinositol lipids but not from the structural lipids. The major fatty acids in the polyphosphoinositides were C-22 carbon polyenoic acids (70%). The significance of these results in relationship to intracellular signalling via inositol phosphates and Ca(2+) is discussed.
For mating Chlamydomonas eugametos gametes to fuse with their partners, they must first lyse part of the anterior cell wall and protrude their mating structures. These responses can be artificially induced by compounds that raise the Cai level, viz. InsP3, A23187, TFP and ethanol. We conclude that calcium should be considered with cAMP to be involved in signal transduction during C. eugametos mating.
In Chlamydomonas eugametos gametes, phosphatidylinositol 4-phosphate (PtdInsP) and phosphatidylinositol 4,5-bisphosphate (PtdInsP2) comprised 0.4 and 0.3% of the whole-cell phospholipids. They were concentrated in the plasma membrane around the cell body and were present in low concentrations in the flagellar membrane. When gametes were fed (32)PO 4 (-) , the label was rapidly incorporated into PtdInsP and PtdInsP2 and only slowly incorporated into structural lipids such as phosphatidylethanolamine and phosphatidylglycerol. Similarly, when a pulse of (32)PO 4 (-) was chased with PO 4 (-) , the label was rapidly lost from the polyphosphoinositol lipids but not from the structural lipids. The major fatty acids in the polyphosphoinositides were C-22 carbon polyenoic acids (70%). The significance of these results in relationship to intracellular signalling via inositol phosphates and Ca(2+) is discussed.
den Ende. H. 1994. Aclin in mating structures of Chtamvdomtmas eugametos gametes. -Phy.siol. Plant. 91: 657-664.The presence of aetin in Chlamydmmmas eugameuis mating structures was studied using monoclonal anii-aetin antibodies. Immunoftuorescent labelling of mating gametes clearly stained their mating structures and this was confirmed al the electron mieroscope level by immunogold labelling of sections. Anti-actin labelling also strongly stained the flagella at the flagellar collar regions and weakEy stained the rest of the flagella. Treatment of gametes wiifi 6-8% ethanol induced mating structures which protruded as large 'balloons'. Balloons stained brilliantly with anti-aetin antibodies and weakly with FITC-phalloidin, a fluorescent reagent Ihat stains F-actin. Isolated mating structure balloons and flagella were analysed using western blouing. A prominent 43 kDa band, eo-migrating vvith actin in erythrocyte ghosts, reacted «ith anti-actin antibodies. The results indicate that aetin is present in mating stiuctures and flagella of both maling types of C. eugametos.
The presence of actin in Chlamydomonas eugametos mating structures was studied using monoclonal anti‐actin antibodies. Immunofluorescent labelling of mating gametes clearly stained their mating structures and this was confirmed at the electron microscope level by immunogold labelling of sections. Anti‐actin labelling also strongly stained the flagella at the flagellar collar regions and weakly stained the rest of the flagella. Treatment of gametes with 6–8% ethanol induced mating structures which protruded as large ‘balloons’. Balloons stained brilliantly with anti‐actin antibodies and weakly with FITC‐phalloidin, a fluorescent reagent that stains F‐actin. Isolated mating structure balloons and flagella were analyzed using western blotting. A prominent 43 kDa band, co‐migrating with actin in erythrocyte ghosts, reacted with anti‐actin antibodies. The results indicate that actin is present in mating structures and flagella of both mating types of C, eugametos.
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