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Synthesis and intracellular transport of protein by the colleterial gland of the cecropia silkmoth 总被引:1,自引:0,他引:1
The tubular portion of the colleterial gland of the cecropia silkmoth secretes protein which labels extensively with glycine and appears as a single peak when separated by acrylamide gel electrophoresis. The radioactive peak does not stain with protein stains or absorb at 280 nm, but is dispersed by Pronase digestion. Synthesis of the peak is sensitive to inhibition by puromycin and cycloheximide but is not sensitive to actinomycin or chloramphenicol inhibition. Amino acid analysis revealed a composition of 30% glycine, 33% aspartic acid, and insignificant amounts of aromatic amino acids; the molecular weight was calculated to be 160,000. Autoradiographic analysis revealed that nearly all the glycine incorporated is transported from the mature secretory cells, and the half-time of secretion is calculated to be 3.4 hr. The possible use of this product as a marker for biochemical differentiation is discussed. 相似文献
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The emergence of the adult Manduca sexta moth is followed by the loss of almost half of this insect's abdominal motoneurons and interneurons (Truman, 1983). This programmed cell death completes the transformation of the nervous system of the caterpillar into that of the moth. The death of these neurons has been previously shown to be a response to an endocrine signal: the decline in ecdysteroids that occurs at the end of metamorphosis (Truman and Schwartz, 1984). Our current research is focussed on the regulation of the fate of a pair of identified motoneurons, the MN-12 cells, in the third abdominal ganglion. Isolation of this ganglion from anterior parts of the nervous system can prevent the death of these cells at the time when they would normally die in response to the decline in ecdysteroids. Transection of the ventral nerve cord at various levels revealed that the source of this regulatory "death signal" is the fused pterothoracic ganglion and that it is transmitted via the interganglionic connectives. We hypothesize that the factors mediating this effect may act in concert with the ecdysteroid decline to specify the exact time of death for individual neurons. 相似文献
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The segmentally arranged Verson's glands are epidermal derivatives comprised of three cells: the duct, saccule, and secretory cells. The development of these glands was followed through the 5th instar and larval-pupal transition of Manduca sexta. The glands are relatively small during the feeding stage, begin to grow at wandering, and undergo about a 50-fold increase in size during the prepupal period. The increase in size is due mainly to the hypertrophy of the secretory cell which synthesizes a heterogeneous set of proteinaceous secretory products. Three prominent 11 to 12 kiloDalton (kD) polypeptides are made by the pharate fifth larval gland, whereas the pupal gland produces polypeptides ranging from 14 to 75 kD with a major complex at 30 to 34 kD. The secretory product is poured out onto the surface of the new cuticle at the time of ecdysis and contains all of the major proteins detected in extracts of the whole gland. The accumulation of secretory products by the gland occurs during the prepupal peak of ecdysteroid and is blocked if this rise is prevented by abdominal isolation. Infusion of 30 micrograms 20-hydroxyecdysone (20-HE) into such isolated abdomens caused synthesis of the pupal products. Treatment with the juvenile hormone mimic, methoprene, during the fifth instar showed that the commitment of the glands to produce the pupal proteins is independent of and occurs before the overlying epidermis becomes committed to make pupal cuticle. 相似文献
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James W. Truman Dorothy B. Rountree Shirley E. Reiss Lawrence M. Schwartz 《Journal of insect physiology》1983,29(12):895-900
The relationship between the ecdysteroid titre and eclosion hormone was explored for the pupal and adult ecdyses of Manduca sexta. Ecdysteroid treatment late during either moult caused a dosedependant delay in the time of ecdysis. Sensitivity to exogenous steroid treatment dropped off as the respective moults neared completion and in both cases coincided with the time of the low point in the endogenous ecdysteroid titre. It was concluded that an ecdysteroid decline is a normal prerequisite for the ecdyses of both stages. The steroid drop is important for two aspects of the eclosion hormone system: it causes target tissues to become sensitive to the peptide and it is a prerequisite for the subsequent release of eclosion hormone itself. Thus, the dual action of the declining ecdysteroid titre insures that when eclosion hormone is released, the tissues will be competent to respond to it. 相似文献
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A cytological and cytochemical examination of Saprolegnia terrestris Cookson was undertaken to resolve the conflict of opinion regarding the life cycle of fungi in the Saprolegniaceae, i.e., whether meiosis is zygotic or gametic. Microspectrophotometric analysis reveals that pre-divisional oogonial and antheridial nuclei are 4C; a subsequent reduction division in the sex organs produces 1C male and female gametes. These division products coexist in the developing oospore and presumably fuse to produce the diploid zygote. Analysis of the somatic hyphal nuclei shows a 2C DNA content, with the characteristic skew and bimodal character indicative of predominantly G1, and some S-period and G2 types. The evidence is completely consistent with gametic meiosis, i.e., a predominant diploid vegetative phase and short haploid phase, and refutes the notion of zygotic meiosis. Further ramifications of the work are discussed along with preliminary results of related work in progress. 相似文献
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Developmental control of xylem hydraulic resistances and vulnerability to embolism in Fraxinus excelsior L.: impacts on water relations 总被引:3,自引:0,他引:3
Welbaum G.E.; Bian D.; Hill D.R.; Grayson R.L.; Gunatilaka M.K. 《Journal of experimental botany》1997,48(3):655-663
The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 104M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar Alaska, an ABA-deficientmutant wil, and its wildtype. Freezinginjury was determined graphically as the temperature that caused50% injury (T50) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T50 for root tissuewas 1C for all genotypes, regardless of treatment. For Alaskashoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by 1.5 to 4.0C, while coldtreatment increased the freezing tolerance by 7.5 to14.8C. Cold treatment increased the freezing toleranceof shoot tips by 9 and 15C for wiland wild-type, respectively. Cold acclimationincreased endogenous ABA concentrations in Alaskashoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of Alaska, respectively.In Alaska shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in wiland wild-type shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization 相似文献
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N-Acetylaspartylglutamate Stimulates Metabotropic Glutamate Receptor 3 to Regulate Expression of the GABAA α6 Subunit in Cerebellar Granule Cells 总被引:1,自引:1,他引:0
Subroto Ghose Barbara Wroblewska Lorenzo Corsi †Dennis R. Grayson ‡Angel L. De Blas Stefano Vicini Joseph H. Neale 《Journal of neurochemistry》1997,69(6):2326-2335
Abstract: We have shown that the vertebrate neuropeptide N-acetylaspartylglutamate (NAAG) meets the criteria for a neurotransmitter, including function as a selective metabotropic glutamate receptor (mGluR) 3 agonist. Short-term treatment of cerebellar granule cells with NAAG (30 µM) results in the transient increase in content of GABAAα6 subunit mRNA. Using quantitative PCR, this increase was determined to be up to 170% of control values. Similar effects are seen following treatment with trans-1-aminocyclopentane-1,3-dicarboxylate and glutamate and are blocked by the mGluR antagonists (2S,3S,4S)-2-methyl-2-(carboxycyclopropyl)glycine and (2S)-α-ethylglutamic acid. The effect is pertussis toxin-sensitive. The increase in α6 subunit mRNA level can be simulated by activation of other receptors negatively linked to adenylate cyclase activity, such as adenosine A1, α2-adrenergic, muscarinic, and GABAB receptors. Forskolin stimulation of cyclic AMP (cAMP) levels abolished the effect of NAAG. The change in α6 levels induced by 30 µM NAAG can be inhibited in a dose-dependent manner by simultaneous application of increasing doses of the β-adrenergic receptor agonist isoproterenol. The increase in α6 mRNA content is followed by a fourfold increase in α6 protein level 6 h posttreatment. Under voltage-clamped conditions, NAAG-treated granule cells demonstrate an increase in the furosemide-induced inhibition of GABA-gated currents in a concentration-dependent manner, indicating an increase in functional α6-containing GABAA receptors. These data support the hypothesis that NAAG, acting through mGluR3, regulates expression of the GABAAα6 subunit via a cAMP-mediated pathway and that cAMP-coupled receptors for other neurotransmitters may similarly influence GABAA receptor subunit composition. 相似文献