Fatty acids and phospholipids of adult and newborn rat hearts and of cultured, neonatal rat heart cells were determined by gas liquid and thin layer chromatographies. In adult heart, the proportion of linoleic acid was higher and that of palmitic acid lower than in newborn hearts or in cultured cells. The relative amounts of linoleic and arachidonic acids in adult heart were affected by the source and amount of dietary fat. In heart cells, after 3 days in culture, the proportion of arachidonic acid resembled that in the newborn and adult rat hearts but showed a gradual and significant decline with age. The gradual shift in fatty acid composition as the cells aged in culture was attributed to outgrowth of mesenchymal cells (fibroblasts and endothelioid cells) characterized by a low relative proportion of arachidonic acid. The amounts of phospholipids in heart cells after 3 days in culture differed from those in the newborn or adult rat hearts. Phosphatidylethanolamine was highest in adult heart (34% of lipid phosphorus) and lowest in cells (26%); lecithin was higher in newborn heart (43%) than in adult heart (37%) or in cells (39%), while sphingomyelin was higher in cells (8%) than in newborn (5%) or adult heart (3%). Phospholipid levels in cultured heart cells were unrelated to those of serum in the growth medium. The absence of a significant change in phospholipid composition after continued incubation of the heart cell cultures for periods up to 3 weeks reflected the major structural role of these lipid components in cell membranes.
High-affinity binding sites (apparent KD 2.87 nM) for [3H]desmethylimipramine ([3H]DMI), have been demonstrated and characterized in membrane preparations of bovine adrenal medulla. The binding of [3H]DMI improved upon pretreatment of the membrane with KCl and was saturable, sodium dependent, and potently inhibited by nisoxetine and imipramine. [3H]DMI binding was also inhibited by various phencyclidine (PCP)- and (or) sigma-receptor ligands, with the following order of potency: haloperidol > rimcazole > (-)-butaclamol > dextromethorphan > MK-801 > (+)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine ((+)-3-PPP) > PCP > N-(2-thienyl)cyclohexyl-3,4-piperidine (TCP) > (+)-SKF-10047 > (-)-SKF-10047. The inhibition produced by sigma ligands was not attributed to stimulation of either sigma 1- or sigma 2-receptors, owing to inactivity of the selective sigma-receptor ligands (+)-pentazocine and 1,3-di(2-tolyl)guanidine (DTG). The inhibition of [3H]DMI binding by sigma- and PCP-receptor ligands was not attributed to PCP1- or PCP2-receptor stimulation, owing to the decreased potency (100-fold) of these ligands in [3H]DMI assays compared with the affinity for brain PCP1 sites, and the ineffectiveness of the PCP2-ligand N-(1-(2-benzo(b)thiophenyl)cyclohexyl)piperidine (BTCP). Scatchard analysis of the inhibition by the sigma-ligands haloperidol and (+)-3-PPP, as well as the PCP1 receptor ligand MK-801, demonstrated noncompetitive interaction with the site bound by [3H]DMI. These studies indicate that bovine adrenomedullary membranes possess a specific receptor for the noradrenaline uptake inhibitor [3H]DMI, which is sensitive to allosteric modulation produced by PCP and sigma-ligands.
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