Molecular cloning of two new human paralogs of 85-kDa cytosolic phospholipase A2

Two new cloned human cDNAs encode paralogs of the 85-kDa cytosolic phospholipase A2 (cPLA2). We propose to call these cPLA2beta (114 kDa) and cPLA2gamma (61 kDa), giving the name cPLA2alpha to the well known 85-kDa enzyme. cPLA2beta mRNA is expressed more highly in cerebellum and pancreas and cPLA2gamma more highly in cardiac and skeletal muscle. Sequence-tagged site mapping places cPLA2beta on chromosome 15 in a region near a phosphoinositol bisphosphate phosphatase. The mRNA for cPLA2beta is spliced only at a very low level, and Northern blots in 24 tissues show exclusively the unspliced form. cPLA2beta has much lower activity on 2-arachidonoyl-phosphatidylcholine liposomes than either of the other two enzymes. Its sequence contains a histidine motif characteristic of the catalytic center of caspase proteases of the apoptotic cascade but no region characteristic of the catalytic cysteine. Sequence-tagged site mapping places cPLA2gamma on chromosome 19 near calmodulin. cPLA2gamma lacks the C2 domain, which gives cPLA2alpha its Ca2+ sensitivity, and accordingly cPLA2gamma has no dependence upon calcium, although cPLA2beta does. cPLA2gamma contains a prenyl group-binding site motif and appears to be largely membrane-bound. cPLA2alpha residues activated by phosphorylation do not appear to be well conserved in either new enzyme. In contrast, all three previously known catalytic residues, as well as one additional essential arginine, Arg-566 in cPLA2alpha, are conserved in both new enzyme sequences. Mutagenesis shows strong dependence on these residues for catalytic activity of all three enzymes.     

Prefrontal cortex lesions impair object-spatial integration

How and where object and spatial information are perceptually integrated in the brain is a central question in visual cognition. Single-unit physiology, scalp EEG, and fMRI research suggests that the prefrontal cortex (PFC) is a critical locus for object-spatial integration. To test the causal participation of the PFC in an object-spatial integration network, we studied ten patients with unilateral PFC damage performing a lateralized object-spatial integration task. Consistent with single-unit and neuroimaging studies, we found that PFC lesions result in a significant behavioral impairment in object-spatial integration. Furthermore, by manipulating inter-hemispheric transfer of object-spatial information, we found that masking of visual transfer impairs performance in the contralesional visual field in the PFC patients. Our results provide the first evidence that the PFC plays a key, causal role in an object-spatial integration network. Patient performance is also discussed within the context of compensation by the non-lesioned PFC.     

Four guaianolides,a eudesmanolide and a germacranolide from Ursinia saxatilis

An investigation of Ursinia saxatilis afforded in addition to known compounds a new eudesmanolide derived from ursialpinolide, a germacranolide rel     

Extremely low frequency resistance fluctuations of chara membrane

The membrane resistivity of resting characean cells was measured and found to fluctuate unpredictably about its resting level. The amplitude of these fluctuations relative to the mean resistivity was normally somewhat larger than the same ratio for the vacuolar voltage; further the resistivity and voltage fluctuations seemed not to be strongly correlated.     

Spontaneous electrical activity in shoots of Ipomoea,Pisum and Xanthium

Summary Extracellular recordings from light-grown shoots of Ipomoea and Xanthium contain trains of spontaneous fluctuations which resemble action potentials. The rise time of the putative action potentials is usually in the range from 1 to 50 ms. Their duration is most frequently between 100 and 400 ms, but they may be either briefer or, in the presence of unusual shoulders, much longer. Their separation interval is typically between 0.1 and 10 s. The trains may last from 1 s to over 2 h.Spontaneous fluctuations also occur individually in shoots of Ipomoea, Xanthium, and Pisum. Although the shapes, rise times, and durations of the individual fluctuations fall within the same range as do those of repetitive fluctuations, a greater number of the individual spikes have a relatively slow time course and a higher apparent amplitude. At times, recordings have periods of activity which appear to result from the superposition of numerous individual fluctuations.There is no evidence that any of the fluctuations propagate, but propagation cannot be excluded on the basis of the extracellular recordings. The fluctuations occur in both dark and light. There is no strong evidence for the occurrence of spontaneous fluctuations in roots.     

Properties of action potentials in Drosera tentacles

Summary Action potentials of Drosera tentacles resemble those of vertebrate peripheral nerves in that they appear to be comprised of relatively uniform spikes, variable shoulders or negative after-potentials, and variable positive after-potentials. The peaking of the spike corresponds to a period of great refractoriness, while action potentials of low amplitude may be fired readily during the negative after-potential. The action potentials fired during the negative after-potential appear to be unlike those of peripheral nerves in that they are of abnormally brief duration. Also apparently different from the case in peripheral nerves is the dependence of the duration of an action potential on the interval separating it from the preceding action potential.Action potentials propagate from the neck of the stalk to its base at about 5 mm s^-1 at room temperature. Propagation may be reversed artificially, consistent with the possibility that the neuroid cells are electrically coupled.     

Influence of phospholipase C on muscarinic acetylcholine receptor binding in rat brain

Treatment of neural membranes from rat cerebral cortex with phospholipase C (phosphatidylcholine cholinephosphohydrolase) inhibited the binding of radiolabelled antagonists to muscarinic acetylcholine receptors. This inhibition was incomplete, was not competitive, and did not appear to be related to the production of inhibitory products. The affinity of carbamylcholine for cortex muscarinic receptors was increased by phospholipase C action. The distribution of receptors between states of high and low affinity was not affected by phospholipase C; rather, the affinity for carbamylcholine of the lowest affinity receptors was selectively increased. This suggests that membrane lipids influence the interaction of the receptor binding subunit with other structures in the synaptic membrane.     

Arabinogalactan protein and wall-associated kinase in a plasmalemmal reticulum with specialized vertices

Summary Arabinogalactan protein and wall-associated kinase (WAK) are suspected to be regulatory players at the interface between cytoplasm and cell wall. Both WAK(s) and arabinogalactan shown likely to represent arabinogalactan protein(s) have been visualized there with computational optical-sectioning microscopy. The arabinogalactan occurs in a polyhedral array at the external face of the cell membrane. WAK, and other proteins as yet unidentified, appear to fasten the membrane to the wall at vertices of the array. Evidence is presented that the array bears an important part of the mechanical stress experienced by the membrane, and it is speculated that the architectural organization of arabinogalactan protein, WAK, and other components of the array is critical for coordination of endomembrane activities, growth, and differentiation. The array has been named the plasmalemmal reticulum.Abbreviations AGP arabinogalactan protein - (-D-Glc)₃ -D-glucosyl Yariv phenylglycoside - (-D-Man)₃ -D-mannosyl Yariv phenylglycoside - SDS-AGE sodium dodecyl sulfate-agarose gel electrophoresis - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - WAK wall-associated kinase