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91.
Helmut Krüger Werner Schr?der Klaus Buchner Ferdinand Hucho 《Journal of Protein Chemistry》1990,9(4):467-473
Inhibition of protein kinase C (PKC) by calmodulin is investigated and we describe the localization of inhibitory sequences within the calmodulin molecule. We present evidence that calmodulin inhibits PKC through an inhibition of the activation of PKC associated with lipid membranes: Binding of PKC to lipid vesicles is not affected, but activation is abolished. The potent calmodulin antagonist R24571 (calmidazol) did not affect the inhibition of PKC by calmodulin at concentrations up to 10–5 M. Two tryptic fragments of calmodulin were isolated which inhibited PKC. They were only slightly less potent than intact calmodulin with an IC50 of 6 µ M compared to 1 µ M of intact calmodulin. They were identified as Ser38-Arg74 and His107-Lys148. Each of the inhibiting fragments contains an intact Ca2+-binding domain with complete helix-loop-helix structure (EF hand). Other calmodulin peptides showed only weak inhibitory activity. Both fragments did not stimulate cAMP phosphodiesterase even at concentrations 100-fold higher than the calmodulin concentration needed for maximal stimulation. None of the fragments acted as a calmodulin antagonist. 相似文献
92.
Summary The ultrastructural and transmitter development of lumbar sympathetic ganglia was studied in embryonic day-6 through-18 chick embryos. At embryonic day 6, ganglia are populated by two morphologically distinct types of neuronal cells and Schwann cell precursors. The neuronal populations basically comprise a granule-containing cell and a developing principal neuron. Granule-containing cells have, an irregularly shaped or oval nucleus with small clumps of chromatin attached to the inner nuclear membrane and numerous large (up to 300 nm) membrane-limited granules. Developing principal neurons display a more rounded vesicular nucleus with evenly distributed chromatin, prominent nucleoli, more developed areas of Golgi complexes, and rough endoplasmic reticulum and large dense-core vesicles up to 120 nm in diameter. There are granule-containing cells with fewer and smaller granules which still display the nucleus typical for granule-containing cells. These granule-containing cells may develop toward developing principal neurons or the resting state of granule-containing cells found in older ganglia. Both granule-containing cells and developing principal neurons proliferate and can undergo degeneration. At embryonic day 9 there are far more developing principal neurons than granule-containing cells. Most granule-containing cells have very few granules. Mitotic figures and signs of cell degeneration are still apparent. Synapse-like terminals are found on both developing principal neurons and granule-containing cells. Ganglionic development from embryonic day 11 through 18 comprises extensive maturation of developing principal neurons and a numerical decline of granule-containing cells. Some granule-containing cells with very few and small granules still persist at embryonic day 18. The mean catecholamine content per neuron increases from 0.044 femtomol at embryonic day 7 to 0.22 femtomol at embryonic day 15. Concomitantly, there is a more than 6-fold increase in tyrosine hydroxylase activity. Adrenaline has a 14% share in total catecholamines at embryonic day 15. Somatostatin levels are relatively high at embryonic day 7 (1.82 attomol per neuron) and are 10-fold reduced by embryonic day 15. Our results suggest the presence of two morphologically distinct sympathetic neuronal precursors at embryonic day 6: one with a binary choice to become a principal neuron or to die, the other one, a granule-containing cell, which alternatively may develop into a principal neuron, acquire a resting state or die. 相似文献
93.
The 76 kD cell-adhesion factor from crayfish haemocytes promotes encapsulation in vitro 总被引:2,自引:0,他引:2
Summary Semigranular cells from the crayfish, Pacifastacus leniusculus, were separated by Percoll gradient centrifugation and were used to study the encapsulation of foreign particles. The semigranular cells were found strongly to encapsulate glass beads coated with haemocyte lysate in which the prophenoloxidase-activating system had been activated with laminarin or with a low concentration of calcium ions. The granular cells only weakly encapsulated these particles. The encapsulationpromoting factor was purified from haemocyte lysates and found to be a 76 kD protein which was recognized by an antiserum to the previously described 76 kD cell-adhesion factor. After the last step in purification (Con A-Sepharose chromatography), the flowthrough consisted of several proteins, which had some, but less, encapsulation-promoting activity and contained a 30 kD band that was also recognized by the antiserum to the 76 kD cell-adhesion factor. If the haemocyte lysate prepared in low [Ca2+] was incubated with a -1,3-glucan prior to purification, no 76 kD protein could be isolated but only a 30 kD protein. The 30 kD protein thus seems to be a degradation product of the 76 kD cell-adhesion factor. We conclude that the 76 kD protein which is released from degranulating haemocytes, and to a lesser extent its 30 kD fragment, can promote encapsulation. Phenoloxidase did not have any encapsulation-promoting activity. 相似文献
94.
Päivi Heikkilä 《Cell and tissue research》1990,259(3):421-427
Summary The present study was undertaken to define the effects of lipoprotein-derived cholesterol and endogenous, de novo synthesized cholesterol on the ultrastructure and function of undifferentiated rat adrenocortical cells [lipoprotein (HDL3 and LDL) receptor-negative, zona glomerulosa-like adrenocortical cells] in primary culture. For this purpose human plasma high density lipoprotein (HDL3) or low density lipoprotein (LDL) was added to culture medium devoid of cholesterol. Steroid secretion remained at the low basal level even after addition of lipoproteins, and the amount of intracellular lipid droplets did not increase. When mevinolin (0.96 µg/ml), an inhibitor of cholesterol synthesis, was added to the culture medium, a low secretion of corticosterone was measured both in serum-free and serum-containing media. Ultrastructurally, lipid droplets disappeared after treatment with mevinolin in both media used. At this concentration of mevinolin cell proliferation was similar to that in the controls, but at higher concentrations (4.8 or 9.6 µg/ml) proliferation was inhibited to 42% and 26% in serum-free medium, and 20% and 12% in serum-supplemented medium, respectively. This study demonstrates that cell proliferation and synthesis of corticosterone by undifferentiated rat adrenocortical cells is identical in the absence or presence of exogenous lipoprotein cholesterol. Inhibition of de novo cholesterol synthesis by mevinolin over a period of 7 days does not inhibit corticosterone secretion or proliferation of cells but decreases the amount of intracellular lipid droplets, thus suggesting utilization of intracellular cholesterol esters. However, higher concentrations of mevinolin inhibit proliferation of cells both in serum-free and serum-containing media. 相似文献
95.
Peter Bräunig 《Cell and tissue research》1990,260(1):95-108
Summary The nervus corporis cardiaci III (NCC III) of the locust Locust migratoria was investigated with intracellular and extracellular cobalt staining techniques in order to elucidate the morphology of neurons within the suboesophageal ganglion, which send axons into this nerve. Six neurons have many features in common with the dorsal, unpaired, median (DUM) neurons of thoracic and abdominal ganglia. Three other cells have cell bodies contralateral to their axons (contralateral neuron 1–3; CN 1–3). Two of these neurons (CN2 and CN3) appear to degenerate after imaginal ecdysis. CN3 innervates pharyngeal dilator muscles via its anterior axon in the NCC III, and a neck muscle via an additional posterior axon within the intersegmental nerve between the suboesophageal and prothoracic ganglia. A large cell with a ventral posterior cell body is located close to the sagittal plane of the ganglion (ventral, posterior, median neuron; VPMN). Staining of the NCC III towards the periphery reveals that the branching pattern of this nerve is extremely variable. It innervates the retrocerebral glandular complex, the antennal heart and pharyngeal dilator muscles, and has a connection to the frontal ganglion.Abbreviations
AH
antennal heart
-
AN
antennal nerves
-
AO
aorta
-
AV
antennal vessel
-
CA
corpus allatum
-
CC
corpus cardiacum
-
CN1, CN2, CN3
contralateral neuron 1–3
-
DIT
dorsal intermediate tract
-
DMT
dorsal median tract
-
DUM
dorsal, unpaired, median
-
FC
frontal connective
-
FG
frontal ganglion
-
HG
hypocerebral ganglion
-
LDT
lateral dorsal tract
-
LMN, LSN
labral motor and sensory nerves
-
LN+FC
common root of labral nerves and frontal connective
-
LO
lateral ocellus
-
MDT
median dorsal tract
-
MDVR
ventral root of mandibular nerve
-
MVT
median ventral tract
-
NCA I, II
nervus corporis allati I, II
-
NCC I, II, III
nervus corporis cardiaci I, III
-
NR
nervus recurrens
-
NTD
nervus tegumentarius dorsalis
-
N8
nerve 8 of SOG
-
OE
oesophagus
-
OEN
oesophageal nerve
-
PH
pharynx
-
SOG
suboesophageal ganglion
-
T
tentorium
-
TVN
tritocerebral ventral nerve
-
VLT
ventral lateral tract
-
VIT
ventral intermediate tract
-
VMT
ventral median tract
-
VPMN
ventral, posterior, median neuron
-
1–7
peripheral nerves of the SOG
-
36, 37, 40–45
pharyngeal dilator muscles 相似文献
96.
Günter Kämper Hans-Ulrich Kleindienst 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1990,167(2):193-200
1. | Filiform hairs of various lengths on the cerci of adult crickets vibrate in a sound field. These movements were measured with a photodetector for sound frequencies from 10 Hz to 200 Hz in the species Acheta domestica, Gryllus bimaculatus and Phaeophilacris spectrum. |
2. | With low air-particle velocities, the hair shafts were deflected sinusoidally from their resting position, without bending or secondary oscillations (Figs. 2 A, 3 A). At higher velocities (from ca. 80 mm/s peak velocity, depending on the properties of the individual hairs), the shaft struck the cuticular rim of the socket in which the base of the hair is seated (Fig. 2B). This contact was made at an average angular displacement from the resting position of 5.16°±1.0°. |
3. | The best frequencies of the hairs were found to be between 40 Hz and 100 Hz (Fig. 5A). The slope of the amplitude curve for constant peak air-particle velocity at frequencies below the best frequencies was between 0 and 6 dB/octave. Long hairs had smaller slope values than short hairs (Fig. 5C). |
4. | At its best frequency the ratio of maximal tip displacement of a hair to the displacement of the air particles in the sound field was between 0.2 and 2. Only a small number of hairs (2 out of 36) showed tip displacements exceeding twice the air-particle displacement. The values of maximal angular displacement were not correlated to hair length (Fig. 5 B). |
5. | The angular displacement of the hairs was phase shifted with respect to the air-particle velocity by 0° to +45° (phase lead) at sound frequencies around 10 Hz and by -45° to -120° (phase lag) at 200 Hz (Figs. 3C, 4B). At a particular frequency long hairs tended to have larger phase lags than shorter hairs (Fig. 5D). |
97.
Dr. Kauko K. Mäkinen Eva Söderling Salam A. Syed Pirkko-Liisa Mäkinen 《Current microbiology》1990,20(1):5-11
The cell extracts of two human oral strains (FN2 and FN3) ofFusobacterium nucleatum displayed exceptionally high-glutamylpeptidase activity as determined withN--l-glutamyl-2-naphthylamine as substrate. This activity was so dominant that the hydrolysis of otherN-aminoacyl-2-naphthylamines progressed at a rate <10% of the former. Two major enzymes (I and II) were partially purified from FN2. I had a molecular weight of 115,000 and did not hydrolyze-glutamylcysteinylglycine (glutathione). II had a molecular weight of 70,000 and rapidly liberated only glutamic acid from glutathione. Strain FN3 contained several enzymes hydrolyzing-glu-2NA. Direct anion exchange chromatography of FN3 cell extracts separated one enzyme that liberated both glutamic acid and glycine from glutathione, one that was inactive against glutathione (but hydrolyzed-glu-2NA), and one that liberated only glutamic acid. Although-glu-2NA was a good synthetic substrate, glutathione was hydrolyzed at least 500 times faster by an enzyme present in both strains. These results indicate that the presence of-glutamylpeptidase activity is very characteristic of theseF. nucleatum strains. 相似文献
98.
The effects of , , and receptor-agonists on forskolin stimulated cyclic adenosine-3, 5-monophosphate (cAMP) formation were examined in astroglial enriched primary cultures from the cerebral cortex of newborn rats. Intracellular cAMP accumulation was quantified by radioimmunoassay. Morphine was used as a -receptor agonist, D-Ala-D-Leu-Enkephalin (DADLE) as a -receptor agonist and dynorphine 1–13 (Dyn) as a -receptor agonist. Basal cAMP levels were unaffected by either the opiate agonists or the antagonists used. In the presence of the cAMP stimulator forskolin, morphine had no significant effect on the cytoplasmic cAMP levels. DADLE caused a dose related inhibition of the forskolin stimulated cAMP accumulation. The effects of this receptor stimulation was blocked with the selective antagonist ICI 174.864. In the presence of Dyn, the forskolin stimulated cAMP accumulation was inhibited in a dose related manner. This receptor stimulation was blocked with the selective antagonist MR 2266. Co-administration of DADLE and Dyn resulted in a non additive inhibition of the forskolin stimulated accumulation of cAMP. These findings indicate that astroglial enriched cultures from the cerebral cortex of rats express and -receptors co-localized ont he same population of cells, and that these receptors are inhibitory coupled to adenylate cyclase. 相似文献
99.
Reinhard Lakes Klaus Kalmring Karl-Heinz Engelhard 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1990,166(4):553-563
Deafferentation experiments during postembryonic development show morphological and/or physiological changes of receptor fibers and of identified auditory interneurons in the CNS of the locusts Locusta migratoria and Schistocerca gregaria after unilateral ablation of one tympanic organ either in the larva or the adult animal.
相似文献
1. | In Locusta migratoria, 5 days after deafferentation, intact, contralateral receptor fibers had sprouted collaterals in the frontal acoustic neuropil of the metathoracic ganglion (Figs. 1, 2). Collateral sprouts were only rarely found in Schistocerca gregaria. |
2. | After about 20 days the deafferented auditory interneurons receive new inputs from the contralateral receptors (Figs. 3, 5, 7, 10). This largely restores their thresholds and intensity/response functions. Collaterals from the first order interneurons cross the midline to the contralateral neuropil (BSN1 neuron, Fig. 4), which is never seen in intact animals. By contrast, in the TN1 neuron no consistent morphological change due to the deafferentation could be found (Fig. 6). |
3. | Interneurons of higher order (AN1, TN3 neuron in locusts) regain their response pattern (Fig. 7) without morphological changes (Fig. 9). Bilateral recordings show that the deafferented interneurons respond more weakly to auditory stimuli than the intact neuron, but the response to vibration stimuli remains unchanged (TN3 neuron, Fig. 8). |
100.
Summary We raised monoclonal antibodies against a membrane fraction ofXenopus neurulae in order to detect tissue-specific cell-surface markers. Here we describe a monoclonal antibody that recognizes an epithelial membrane-associated antigen (EMA) in immunohistological stainings. The tissue-specific and membrane-associated antigen detected in immunohistological stainings could serve as useful marker in epithelium differentiation and membrane organization of the early embryo. In tadpoles and adults EMA was found in specific epithelial tissues derived from different germ layers such as kidney, skin, gut, pancreas, epiphysis and choroid plexus. In the cleaving embryo this antibody stained newly formed membranes between blastomeres from the two-cell stage onwards. Cytoplasmic staining in large oocytes and early embryos was also observed. The possibility that the cytoplasmic signal represents a maternal store of membrane material is discussed. 相似文献