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1.
Summary The superior cervical ganglia (SCG), celiac superior mesenteric ganglia (CMG), and splanchnic nerve of unoperated guinea pigs, as well as both proximal and distal stumps of a previously transected branch of the postganglionic plexus of the CMG, were immunostained for somatostatin (SS). In addition, the PAP technique was adapted for fine-structural visualization of SS. A greater proportion of cells were labeled for SS in the CMG than in the SCG. PAP molecules were present in one type of intraganglionic axons. Only two labeled axons were found in the splanchnic nerve. Neither proximal nor the distal stump of the transected CMG postganglionic nerve contained labeled axons. The present results support the hypothesis that the intraganglionic axons labeled for SS arise from SS-containing intraganglionic neurons.  相似文献   

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[125I]-Nerve growth factor (NGF) binding was studied in superior cervical ganglia from mice 4-7 months and 24-27 months of age. Scatchard analysis demonstrated losses of both high and low affinity components of NGF receptor. These results indicate loss of NGF receptors may lead to the diminished responsiveness to NGF in aged sympathetic ganglion neurons.  相似文献   

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The nerve growth factor (NGF) receptor from microsomes of adult rabbit superior cervical ganglia has been solubilized with Triton X-100 and sodium deoxycholate. The physical properties of the detergent-extracted NGF receptor were assessed by Sepharose 6B chromatography and sucrose density gradient ultracentrifugation studies in H2O and D2O. The predominant form of the NGF receptor has a Stokes radius of 71 A, a partial specific volume of 0.74 ml/g, a sedimentation coefficient of 4.3 S, and a frictional ratio of 1.8. From these parameters, it can be calculated that the NGF receptor in Triton X-100 is a minimally hydrophobic, highly asymmetric, intrinsic membrane protein with a molecular weight of approximately 135,000. A form of the receptor with a sedimentation coefficient of 10.4 S was occasionally seen which appears to represent an aggregated form of the 4.3 S moiety.  相似文献   

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Ribonuclease activity at pH 7.1 (alkaline ribonuclease) was determined in homogenates of rat superior cervical ganglion up to 5 days after postganglionic nerve injury under optimal conditions of assay. Measurements were performed in the presence and absence of the sulfhydryl blocking agent, N-ethylmaleimide, to assess the proportion of alkaline ribonuclease apparently bound to endogenous inhibitor. Total ribonuclease activity per ganglion was stimulated 1.3 fold by 1 day after injury and remained elevated over the 5 day period. Free ribonuclease activity accounted for about 60% of the observed increase in total activity at day 1, but had returned to control level by day 3. At day 3 the entire 90% increase in total activity was attributable to ribonuclease bound to endogenous inhibitor (i.e. latent activity). These changes are occurring at times after nerve injury when marked alterations in RNA turnover have been observed, implicating alkaline ribonucleases in the control of RNA metabolism during nerve regeneration.  相似文献   

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Under study was the influence of the nerve growth factor on morpho-functional and cytometrical characteristics of sympathetic neurons after cutting their axons. The work was carried out in albino rats which were subjected to dissection of the external and internal carotid nerves of the cranial cervical ganglion. This operation resulted in death of about 50% of the neurons of the node in question, in 30 and 60% in reduction of the intensity of fluorescence of the neuron body catecholamines within 7 and 14 days. The cause of these changes might be the cessation of entrance of the nerve growth factor from target tissues by means of retrograde transport. After 6-10 exogenous administrations of the latter (10-20 mkg/g of the body weight) 14 days after operation the amount of neurons in the cranial cervical ganglion did not practically differ from that in the group of intact rats. Seven and fourteen days later specific luminescence of the neuron body catecholamines was 24-22% greater than the values found in the operated animals given the isotonic solution of sodium chloride. Parallel stimulation of the formation of a powerful network of collateral branches by injured axons was performed resulting in more considerable (3-fold) increase of the ganglion mass than in the operated animals (2-fold) not treated with a neuro-growth protein. The newly formed fibers had not only an intercellular but also subcapsular arrangement which spoke of higher reparative--regenerative potencies of the neuron.  相似文献   

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The predominant source of nerve growth factor (NGF) used by mature sympathetic neurons originates in their target organs (Heumann, R., Korsching, S., Scott, J., and Thoenen, H. (1984), EMBO J. 3, 3183-3189; Korsching, S., and Thoenen, H. (1985), J. Neurosci. 5, 1058-1061). We have determined the NGF content of two sympathetically innervated mouse organs, submandibular gland and heart ventricle, and of sympathetic ganglia from mouse and rat between embryonic Day 12 (E12) and adulthood. NGF levels were measured by a two-site enzyme immunassay (Korsching, S., and Thoenen, H. (1983), Proc. Natl. Acad. Sci. USA 80, 3513-3516). In heart ventricle and submandibular gland, NGF first became detectable around the time of initial innervation by sympathetic neurons (E12 and E13, respectively) and increased respectively 14- and 7-fold in the following 2 days, to reach adult levels already at E14 for heart ventricle (1.4 +/- 0.2 ng NGF/g wet wt). NGF in the superior cervical ganglion (SCG) was first detected at the same time as in its target organ, the submandibular gland. NGF content in the SCG then increased 6-fold during the next 2 days and continued to increase until the end of the third postnatal week, when adult levels were reached. Although the levels of NGF in the adult mouse submandibular gland are sexually dimorphic and six orders of magnitude higher than those in other sympathetic target organs, no sex difference in the NGF content was found in either developing submandibular gland or SCG until the end of the third postnatal week. Moreover, the steep NGF increase observed in the male submandibular gland after postnatal Day 18 (250-fold within the following 3 days and up to the 55,000-fold in the next 7 days) was not reflected in a corresponding increase in the NGF content of the male SCG. These data indicate that, in accordance with earlier findings (see Levi-Montalcini, R., and Angeletti, P. U. (1968), Physiol. Rev. 48, 534-569), SCG neurons do not have access to the large amounts of NGF synthesized during and after adolescence in the mouse submandibular gland. Our results support the concept that initial fiber outgrowth of sympathetic neurons is neither dependent on NGF nor mediated by it. The time course of NGF levels in the SCG is consistent with the concept that sympathetic neurons are provided with NGF by means of retrograde axonal transport from the innervated organs already early in development.  相似文献   

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Summary The fine structure of mitochondrial accumulations in axonal swellings of human sympathetic ganglia is described. A typical swelling contained, in addition to regularly organized mitochondria, bundles of neurofilaments and vesicles as well as large dense-cored vesicles and myelin figures. Synaptic contacts between axonal swellings with mitochondrial accumulations and ganglion cells were not found.A three-dimensional model of the mitochondrial accumulation based on serial sectioning is presented. The possible degenerative and regenerative features of these accumulations are discussed. It is possible that mitochondrial accumulations are functionally active energy producers rather than results of degenerative processes.  相似文献   

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The embryonic mouse superior cervical ganglion (SCG) in culture was employed to define the role of ongoing metabolic processes in morphological and biochemical development. The 14 gestational day SCG does not require added nerve growth factor (NGF) for differentiation in vitro. Consequently, its use allows study of intraganglionic regulation of neuronal growth in the absence of complicating, exogenous growth factors. Ganglia were cultured without added NGF, in medium containing various metabolic inhibitors; neurite elaboration and development of tyrosine hydroxylase (T-OH) activity, a biochemical marker of adrenergic maturation, were evaluated. Neurite elaboration proceeded normally with inhibition of RNA synthesis by actinomycin D, or of protein synthesis by cycloheximide or puromycin. In contrast, inhibition of RNA or protein synthesis prevented normal development of T-OH activity. However, neurites and T-OH developed normally in the presence of DNA synthesis inhibition by cytosine arabinoside, which markedly reduced the nonneuronal cell population. These observations suggest that neurite elaboration and the ontogenetic increase in T-OH activity are regulated differently in ganglia cultured in the absence of exogenous NGF. Moreover, the initial outgrowth of neurites and increase in T-OH activity may occur independent of peripherally migrating support cells in this embryonic ganglion.  相似文献   

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Presley  R.  Santer  R. M.  Lu  K.-S.  Lever  J. D. 《Cell and tissue research》1977,178(1):49-60
Summary Chicken embryo sympathetic ganglion fragments, grown in culture for three days, were incubated for short periods in appropriate experimental and control media to provide information on axonal uptake of exogenous noradrenaline in the presence and absence of the uptake-blocking drugs, protriptyline and phenoxybenzamine. The cultures were dried, gassed in formaldehyde vapour and examined by fluorescence microscopy. Spot readings were taken on axon-bundles using a microspectrophotometer. Readings were corrected for the amount of tissue present in each field, using graticule and interferometric methods. Brightness was uniform along individual axon-bundles, but varied from bundle to bundle within the same culture. Despite this range in brightness the readings fell into a normal distribution about the mean. Dose-dependent increases in brightness were obtained following incubation with noradrenaline. A clear indication of the effect of the blocking drugs was observed but the wide range in brightness prevented reliable quantitative data on these effects from being obtained. It was concluded that the wide range of brightness within each culture reflected a lack of equivalence in the intrinsic noradrenaline-uptake properties of the axon-bundles developing in such cultures.This work was supported by a grant from the Medical Research Council. We wish to thank Mrs. G. O'Shea for valuable technical assistance, and Messrs. P.F. Hire and K. Twohigg for help with the illustrations. Phenoxybenzamine was a gift from Smith, Kline and French Ltd.  相似文献   

14.
Sympathetic neurons depend on nerve growth factor (NGF) for their survival both in vivo and in vitro. In culture, the neurons die after NGF withdrawal by an autonomous cell death program but whether these neurons die by apoptosis is under debate. Using vital DNA stains and in situ nick translation, we show here that extensive chromatin condensation and DNA fragmentation occur before plasma membrane breakdown during the death of NGF-deprived rat sympathetic neurons in culture. Furthermore, kinetic analysis of chromatin condensation events within the cell population is consistent with a model which postulates that after NGF deprivation nearly all of the neurons die in this manner. Although the dying neurons display membrane blebbing, cell fragmentation into apoptotic bodies does not occur. Apoptotic events proceed rapidly at around the time neurons become committed to die, regardless of neuronal culture age. However the duration of NGF deprivation required to commit neurons to die, and the rate at which apoptosis occurs, increase with culture age. Thus, within the first week of culture, apoptosis is the predominant form of cell death in sympathetic neurons.  相似文献   

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Summary The effects of chronic lesions of rat lumbar spinal or sciatic nerves on the binding of Glycine max (soybean) agglutinin to galacto-conjugates, in small-and medium-size primary sensory neurons of the L4 and L5 dorsal root ganglia, were examined over a 580-day period. Spinal nerve section resulted in a marked decrease in the population of stained neurons within 7 days. However, despite some retrograde morphological changes triggered by axonal injury, the proportion of stained nerve cells was normalized 180 days postoperatively. This temporary decrease in perikaryal lectin reactivity was initially associated with a marked accumulation of stained material in the nerve, proximal and distal to the site of section, with similar accumulations also being noticeable at each level of injury in sciatic nerves subjected to double ligature. This may reflect the presence of glycocompounds linked to the autolysis of nerve fibers during the phase of retrograde dying-back and Wallerian degeneration. At later stages, stained deposits could be seen scattered along central and peripheral axonal processes of the dorsal root ganglion neurons in the vicinity of the cell body. They may indicate a disturbance in the peripheral turnover of glycoproteins in chronically-transected nerves, with piling up of neuronal products. Sciatic nerve injury caused similar but less severe effects which, except for the L4 ganglion cells, were rapidly reversible.  相似文献   

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