Stimulus-response coupling in marine sponge cell aggregation: lipid metabolism and the function of exogenously added arachidonic and docosahexaenoic acids.
Cells of the marine sponge, Microciona prolifera, the most ancient of the animal cells which clump on recognition, resemble neutrophils and platelets in undergoing stimulus-response coupling when exposed to Ca2+ ionophores and phorbol esters. We have studied lipid content and remodelling in sponge cells by thin-layer, gas-liquid, and high-performance liquid chromatography (HPLC) analyses supplemented by ultraviolet and mass spectroscopy. Phosphatidylcholine (PC) (35.6%), phosphatidylethanolamine (PE) (27.4%) and phosphatidylserine (PS) (21.4%) constituted the bulk of phospholipids detected. The major fatty acids were all polyenoic; 22:6 (22%), 26:2 (17%) and 26:3 (15%). Arachidonic acid (20:4), present as 2.7% of total phospholipid, and docosahexanoic acid (22:6) were found to elicit aggregation of sponge cells when added (10 microM) in synergy with ionomycin (1 microM), resembling in their effects those of phorbol esters (but not phorbol) and 1-oleyl-2-acetylglycerol (OAG). Moreover, 20:4 and 22:6, as well as phorbol ester and OAG, overcame the block to aggregation imposed by colchicine and vinblastine. Kinetic studies of lipid remodelling showed that aggregating cells diverted [14C]22:6 or [14C]20:4 from triacylglycerol into diacylglycerol and phospholipids; appearance of label in phosphatidic acid and phosphatidylinositol (PI) anteceded labeling of phosphatidylcholine. In unstimulated cells, [14C]22:6 was rapidly incorporated into phosphatidylcholine with little accumulation in phosphatidate. Although 22:6 and 20:4 resembled OAG and phorbol esters in overcoming the effects of colchicine and vinblastine (which had no effects on overall lipid metabolism), they did not reverse the block to aggregation of nordihydroguaiaretic acid (NDGA) (which inhibited lipid metabolism). Under none of these circumstances was 22:6 or 20:4 converted to cyclooxygenase or lipoxygenase products in the course of aggregation: all labeled acyl groups remained present as unmodified fatty acids on alkaline hydrolysis. These data not only extend the observations of Muller et al. (J. Biol. Chem. 262 (1987) 9850-9858) on the role of phosphoinositides and C kinase in marine sponge cell aggregation, but also demonstrate that sponges form diacylglycerols in the process. We suggest that exogenous 22:6 and 20:4 (like phorbol esters or OAG) can substitute for endogenous diacylglycerol in the activation of protein kinase C.
Weissmann
G
G
Marine Biological Laboratory, Woods Hole, MA.
Riesen
W
W
Davidson
S
S
Waite
M
M
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002449:Cell Aggregation / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003078:Colchicine / Q000494:pharmacology; D004281:Docosahexaenoic Acids / Q000494:pharmacology*; D004987:Ethers / Q000494:pharmacology; D005227:Fatty Acids / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry; D015759:Ionomycin; D050356:Lipid Metabolism; D009637:Masoprocol / Q000494:pharmacology; D010703:Phorbol Esters / Q000494:pharmacology; D010743:Phospholipids / Q000032:analysis; D011161:Porifera / Q000187:drug effects / Q000502:physiology*; D014747:Vinblastine / Q000494:pharmacology
D001095:Arachidonic Acids; D004987:Ethers; D005227:Fatty Acids; D010703:Phorbol Esters; D010743:Phospholipids; D004281:Docosahexaenoic Acids; D016718:Arachidonic Acid; D015759:Ionomycin; D014747:Vinblastine; D009637:Masoprocol; D003078:Colchicine; D002118:Calcium