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1.
The ultrastructural properties of orexin 1-receptor-like immunoreactive (OX1R-LI) neurons in the dorsal horn of the rat spinal cord were examined using light and electron microscopy techniques. At the light microscopy level, the most heavily immunostained OX1R-LI neurons were found in the ventral horn of the spinal cord, while some immunostained profiles, including nerve fibers and small neurons, were also found in the dorsal horn. At the electron microscopy level, OX1R-LI perikarya were identified containing numerous dense-cored vesicles which were more heavily immunostained than any other organelles. Similar vesicles were also found within the axon terminals of the OX1R-LI neurons. The perikarya and dendrites of some of the OX1R-LI neurons could be seen receiving synapses from immunonegative axon terminals. These synapses were found mostly asymmetric in shape. Occasionally, some OX1R-LI axon terminals were found making synapses on dendrites that were OX1R-LI in some cases and immunonegative in others. The synapses made by OX1R-LI axon terminals were found both asymmetric and symmetric in appearance. The results provide solid morphological evidence that OX1R is transported in the dense-cored vesicles from the perikarya to axon terminals and that OX1R-LI neurons in the dorsal horn of the spinal cord have complex synaptic relationships both with other OX1R-LI neurons as well as other neuron types. 相似文献
2.
Guan JL Wang QP Kageyama H Kita T Takenoya F Hori T Shioda S 《Regulatory peptides》2005,129(1-3):17-23
The distribution of orexin A immunoreactivity and the synaptic relationships of orexin A-positive neurons in the rat area postrema were studied using both light and electron microscopy techniques. At the light microscope level, numerous orexin A-like immunoreactive fibers were found within the area postrema. Using electron microscopy, immunoreactivity within fibers was confined primarily to the axon terminals, most of which contained dense-cored vesicles. Both axo-somatic and axo-dendritic synapses made by orexin A-like immunoreactive axon terminals were found, with these synapses being both symmetric and asymmetric in form. Orexin A-like immunoreactive axon terminals could be found presynaptic to two different immunonegative profiles including the perikarya and dendrites. Occasionally, some orexin A-like immunoreactive profiles, most likely to be dendrites, could be seen receiving synaptic inputs from immunonegative or immunopositive axon terminals. The present results suggest that the physiological function of orexin A in the area postrema depends on synaptic relationships with other immunopositive and immunonegative neurons, with the action of orexin A mediated via a self-modulation feedback mechanism. 相似文献
3.
Dr. F. Hajós 《Cell and tissue research》1980,206(3):477-486
Summary Synaptic clefts of rat cerebral and cerebellar axodendritic spine synapses were studied after aldehyde-perfusion and subsequent immersion into osmic acid or after processing by the aldehyde-PTA technique. The threedimensional examination of aldehyde-perfused, osmic acid postfixed synapses revealed a double-layered intracleft lamina comparable in dimensions and position to the cleft density of non-osmicated, PTA-stained synapses. The relationship of this lamina to perisynaptic astroglial processes was pointed out. Densitometric analysis of the cleft area suggested the identity of the intracleft lamina of osmicated synapses with the cleft density of non-osmicated, PTA-stained synapses. 相似文献
4.
Mixed-rod cone bipolar (Mb) cells of goldfish retina have large synaptic terminals (10 mum in diameter) that make 60-90 ribbon synapses mostly onto amacrine cells and rarely onto ganglion cells and, in return, receive 300-400 synapses from gamma-aminobutyric acid (GABA)-ergic amacrine cells. Tissue viewed by electron microscopy revealed the presence of double-membrane-bound processes deep within Mb terminals. No membrane specializations were apparent on these invaginating processes, although rare vesicular fusion was observed. These invaginating dendrites were termed "InDents". Mb bipolar cells were identified by their immunoreactivity for protein kinase C. Double-label immunofluorescence with other cell-type-specific labels eliminated Müller cells, efferent fibers, other Mb bipolar cells, dopaminergic interplexiform cells, and somatostatin amacrine cells as a source of the InDents. Confocal analysis of double-labeled tissue clearly showed dendrites of GABA amacrine cells, backfilled ganglion cells, and dendrites containing PanNa immunoreactivity extending into and passing through Mb terminals. Nearly all Mb terminals showed evidence for the presence of InDents, indicating their common presence in goldfish retina. No PanNa immunoreactivity was found on GABA or ganglion cell InDents, suggesting that a subtype of glycine amacrine cell contained voltage-gated Na channels. Thus, potassium and calcium voltage-gated channels might be present on the InDents and on the Mb terminal membrane opposed to the InDents. In addition to synaptic signaling at ribbon and conventional synapses, Mb bipolar cells may exchange information with InDents by an alternative signaling mechanism. 相似文献
5.
Electron microscopic examination of the orexin immunoreactivity in the dorsal raphe nucleus 总被引:1,自引:0,他引:1
The ultrastructure and the synaptic relationships of the orexin-A-like immunoreactive fibers in the dorsal raphe nucleus were examined with an immunoelectron microscopic method. At the electron microscopic level, most of the immunoreactive fibers, a varicosity appearance at the light microscopic level, were found as axon terminals. The large dense-cored vesicles contained in the immunoreactive axon terminals were the most intensely immunostained organellae. These axon terminals were often found to make synapses. While the axo-dendritic synapses were usually asymmetric in appearance, the axo-somatic synapses were symmetric. Orexin-A-like immunoreactive processes with no synaptic vesicles were also found. These processes often received asymmetric synapses. With less frequency, the synapses were found between the orexin-like immunoreactive processes. The results suggest that the orexin peptides are stored in the large dense-cored vesicles; the orexin-containing fibers may have influences on the physiological activities of the dorsal raphe nucleus through direct synaptic relationships. 相似文献
6.
Ultrastructural localization of RFamide-like peptides in neuronal dense-cored vesicles in the peduncle of Hydra 总被引:2,自引:0,他引:2
O Koizumi J D Wilson C J Grimmelikhuijzen J A Westfall 《The Journal of experimental zoology》1989,249(1):17-22
The presence of Arg-Phe-amide (RFamide)-like peptides in dense-cored vesicles in neurons of the peduncle of Hydra was demonstrated by immunogold electron microscopy. Thin sections of Lowicryl-embedded tissue labeled with antisera to RFamide and 5-nm gold-conjugated, secondary antibody and of Epon-Araldite-embedded tissue labeled with 15-nm gold particles revealed a concentration of RFamide-like immunoreactivity over the granular cores of vesicles in epidermal ganglion cells. Gold-labeled, dense-cored vesicles were present in the perikaryon, long thin neurites, and axon terminals of these neurons. The aggregation of labeled dense-cored vesicles in an axon terminal on the myoneme of an epitheliomuscular cell suggests a possible function of RFamide-like peptides in neuromuscular transmission. Gold staining of dense-cored vesicles completely disappeared when the RFamide antiserum was preabsorbed with 10 micrograms/ml RFamide. These results are the first demonstration that the dense-cored vesicles of Hydra neurons contain a neuropeptide. 相似文献
7.
Summary Ultrastructural post-embedding immuno-gold techniques were applied to the supraoptic nucleus and the neurohypophysis of mice and rats. The primary antibodies were three different monoclonal antineurophysins, used in protein A-gold and immunoglobulin-gold procedures. Conventional plastic embedding as well as hydrophilic media (L.R. White) were used; non-osmicated and osmicated tissues were immunolabeled; sodium metaperiodate oxidation was used, but was not essential for immunolabeling.Vasopressinergic and oxytocinergic NSGs were identified by the specific immunoreactivity of their respective neurophysins on adjacent thin sections, and by sequential double labeling on the same thin section using two different antibodies associated with gold probes of different diameters. The immunoidentification indicates that vasopressin NSGs can additionally be differentiated as larger, with more electron-dense matrix, and susceptible to damage by sodium metaperiodate.The only organelles consistently labeled were neurosecretory granules (NSGs), either intact or within lysosomal configurations. Some lysosomal dense bodies were immunoreactive even when discrete NSGs were no longer morphologically recognisable within them. Labeled NSGs were located within neuronal cell bodies, along axonal shafts and within axonal swellings and endings; occasionally immunoreactive NSGs were observed within synaptic boutons. Labeling intensity was semi-quantitatively gauged by counting gold particles in relation to numbers of NSGs per axonal varicosity.The precise localisation achieved with particulate immunogold labeling surpasses that previously obtained with diffuse electron-dense immunoreaction products. 相似文献
8.
Guedes RP Marchi MI Viola GG Xavier LL Achaval M Partata WA 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2004,138(1):19-28
Using immunohistochemistry and optical densitometry, somatostatin (SOM), calcitonin gene-related peptide (CGRP), and gamma-aminobutyric acid (GABA) were investigated in the lumbosacral spinal cord of the frog Rana catesbeiana after sciatic nerve transection. In control animals, the densest network of the SOM-, CGRP- and GABA-like immunoreactive fibers was located in the dorsal part of the lateral funiculus. SOM and GABA-like fibers were found in the dorsal terminal field and in the mediolateral band. The latter region showed CGRP and SOM-like immunoreactive cell bodies. SOM- and GABA-like immunoreactive neurons also occurred around the cavity of the central canal, and other GABA-like fibers were found in the ventral terminal field. While the ventral horn showed scarce somatostatin-like fibers, the putative motoneurons were immunoreactive for the two peptides investigated and GABA, but only a few SOM- and GABA-like fibers occurred in the ventral funiculus. After axotomy, GABA-like immunoreactivity decreased in the dorsal part of the lateral funiculus on the same side of the lesion. The other regions remained labeled. These changes were observed at 3 days following axonal injury and persisted at 5, 8 and 15 days. There was no significant difference in the pattern of CGRP- and SOM- immunoreactivity between the axotomized and the control sides. These results are discussed in relation to the effects of the peripheral axotomy on GABA, SOM, and CGRP expression in vertebrates, emphasizing the use of frogs as a model to study the effects of peripheral nerve injury. 相似文献
9.
M M Silver S A Hearn L D Lines M Troster 《The journal of histochemistry and cytochemistry》1988,36(8):1031-1036
We used a post-embedding immunoelectron microscopy method, using protein A-gold, to detect calcitonin and chromogranin A immunoreactivity in three cases of human medullary thyroid carcinoma. Because the epoxy-embedded tissue had been fixed (glutaraldehyde or formaldehyde) and osmicated before embedment, the proteins were identified in optimally preserved tissue. Uranyl and lead staining was used after immunolabeling, so that the tissue was also optimally contrasted. The morphological advantage provided by osmication was tested by labeling rat thyroid gland C-cells for calcitonin. The protein A-gold technique allowed localization of both antigens to the contents of membrane-bound secretory granules in the tumor cells. In one case, labeling density for each antigen was measured over several intercellular compartments and the interstitium. Calcitonin, but not chromogranin A, reactivity was also identified in intracellular amyloid fibrils in two cases, showing that the constant region of calcitonin is preserved in amyloid deposits related to the tumor cells. 相似文献
10.
Guan, J.-L., Q.-P. Wang and Y. Nakai. Electron microscopic observation of delta-opioid receptor-1 in the rat area postrema. Peptides 18(10) 1623–1628, 1997.—The ultrastructural localization of delta-1-opioid-receptor in the rat area postrema was quantitatively studied by pre-embedding avidin-biotin-peroxidase-complex technique. Most of the immunoreactive profiles (67.4%) observed in the present study were axon terminals, whereas the immunopositive dendrites were less (28.3%). Within the axon terminals, the immunoreactivity was found stronger in the dense-cored vesicles than in the small, clear, and round vesicles. Almost 2/3 of the DOR-1 immunoreactive axon terminals had DAB reacted dense-cored vesicles. About half of the immunopositive axon terminals were found to make synapse to dendrites. The dendrites postsynaptic to DOR-1 immunoreactive axon terminals were identified as DOR-1 immunoreactive or not, mainly according to the immunoreactive appearance of the postsynaptic membrane. About half of the DOR-1 immunoreactive dendrites were observed to receive synapse; most of them have their immunoreactivity results at the postsynaptic membranes. 相似文献
11.
Scanning and transmission electron microscopy of the pharynx of the sea anemone Aiptasia pallida revealed a heavily ciliated epidermis and two types of gland cells not known previously to be innervated. By tracing serial cross sections of the pharynx, we located and characterized two types of neuroglandular synapses (i.e., those having clear vesicles and those with dense-cored vesicles). The diameters of the vesicles at each synapse were averaged; clear vesicles ranged from 70 to 103 nm in diameter and were observed at synapses to both mucous and zymogenic gland cells. Dense-cored vesicles ranged from 53 to 85 nm in diameter and were observed at synapses to two mucous gland cells. One mucous gland cell had three neuroglandular synapses, one with clear vesicles and two with dense-cored vesicles. The occurrence of either clear or dense-cored vesicles at neuroglandular synapses suggests that at least two types of neurotransmitter substances control the secretion of mucus in the sea anemone pharynx. To date, only clear vesicles have been observed at a neurozymogenic gland cell synapse in the pharynx. No evidence of immunoreactivity to phenylethanolamine-N-methyl transferase was observed at neuroglandular synapses, suggesting that adrenaline is not a transmitter in the pharynx of A. pallida. 相似文献
12.
N P Veselkin V O Adanina J P Rio J Repérant 《Rossi?skii fiziologicheski? zhurnal imeni I.M. Sechenova / Rossi?skaia akademiia nauk》1999,85(4):515-522
Using the electron microscopy immunocytochemistry, the GABA and glycine immunoreactivity was studied in presynaptic axon terminals of the spinal cord central gray in the lamprey Lampetra fluviatilis. All immunopositive presynaptic terminals contacting motoneurones or non-identified post-synaptic profiles were divided into only GABA- (44%), only glycine-immunopositive terminals (26%), and both GABA- and glycine-containing terminals (30%). The glycine-immunopositive axon terminals contained flattened synaptic vesicles. Large dense core vesicles were co-localised with conventional synaptic vesicles in 74% of GABA-containing presynaptic terminals. 相似文献
13.
Axonal agranular reticulum and synaptic vesicles in cultured embryonic chick sympathetic neurons 总被引:10,自引:6,他引:4
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Cultured chick embryonic sympathetic neurons contain an extensive axonal network of sacs and tubules of agranular reticulum. The reticulum is also seen branching into networks in axon terminals and varicosities. The axonal reticulum and perikaryal endoplasmic reticulum resemble one another in their content of cytochemically demonstrable enzyme activities (G6Pase and IDPase) and in their characteristic membrane thicknesses (narrower than plasma membrane or some Golgi membranes). From the reticulum, both along the axon and at terminals, there appear to form dense-cored vesicles ranging in size from 400 to 1,000 Å in diameter. These vesicles behave pharmacologically and cytochemically like the classes of large and small catecholamine storage vesicles found in several adrenergic systems; for example, they can accumulate exogenous 5-hydroxydopamine. In addition, dense-cored vesicles at the larger (1,000 Å) end of the size spectrum appear to arise within perikaryal membrane systems associated with the Golgi apparatus; this is true also of very large (800–3,500 Å) dense-cored vesicles found in some perikarya. 相似文献
14.
Dr. Y. Ibata Y. Nojyo T. Matsuura H. Yoshikawa Y. Sano 《Cell and tissue research》1975,160(2):139-153
The arcuate nucleus of normal cats and of cats treated with 5-hydroxydopamine (5-OHDA) was investigated by electron microscopy. The neurons of the arcuate nucleus were classified into three types, clear, intermediate and dark, according to their fine structure. The clear type contained numerous dense-cored vesicles and well developed cell organelles. All three types were frequently seen to be partially surrounded by glial processes. Many axo-somatic and axo-dendritic synapses mostly small in diameter were also observed around the neurons. Synaptic contacts were demonstrated between axon endings and axonal processes which contained elementary granules. After administration of 5-OHDA small and large dense-cored vesicles appeared in the nerve endings surrounding the neurons. The relationship between the dense-cored vesicles in the perikarya and dopamine was briefly discussed. 相似文献
15.
Our previously published studies are here reviewed detailing neuro-cnidocyte synapses, demonstrating putative neurotransmitter
substances, and identifying complex neural pathways in sea anemones. Synapses were traced to their contacts on nematocytes
and spirocytes by transmission electron microscopy of serial thin sections of tentacles. In five animals, cells containing
microbasic p-mastigophores had synapses with clear vesicles, whereas cells containing basitrichous isorhizas had synapses with dense-cored
vesicles, providing preliminary evidence for a selectivity of neurotransmitter types for different nematocysts. Either clear
or dense-cored synaptic vesicles were also present at neuro-spirocyte contacts. Antho-RFamide immunoreactivity occurred in
some anthozoan synaptic vesicles and immunogold labeling of serotonin was found at a neuro-spirocyte synapse. Neural pathways
included direct innervation of spirocytes by sensory cells, sequential neuro-neuro-spirocyte and neuro-neuro-nematocyte synapses
and reciprocal synapses involving axons of both sensory cells and ganglion cells. Such synaptic patterns resemble neuro-effector
pathways found in higher animals and lay to rest the independent effector hypothesis for cnidocyte discharge in tentacles
of sea anemones. 相似文献
16.
目的 观察鸡脊髓背角胶状质中calbindin-D28k(CB)阳性终末的超微结构及其与含有substance P(SP)中央末梢之间的联系.方法 应用免疫电镜技术观察鸡脊髓背角胶状质中CB阳性终末的超微结构,并应用激光共聚焦显微镜观察鸡脊髓背角胶状质中CB和SP阳性突触小球中央末梢之间的关系.结果 电镜下观察:1)突触小球中含有心小泡的中央末梢呈CB免疫阳性;2)突触小球内或外的部分含小泡的树突呈CB免疫阳性;以及3)突触小球外的部分轴突呈CB免疫阳性.在突触结构内,CB免疫阳性反应物主要分布于突触后膜上.免疫荧光双标记法显示,SP阳性的含有心小泡的中央末梢呈CB阳性.结论 突触小球的中央末梢中CB与SP共存,提示CB可能通过其钙离子缓冲作用,参与脊髓的痛觉调制. 相似文献
17.
Summary The ultrastructure of the nerves forming the submucous plexus of cervical and thoracic parts of the trachea was studied in the guinea-pig. Specimens were obtained from 6 animals perfused with warm fixative and from 6 animals in which pieces of trachea were incubated in buffer containing 5-hydroxydopamine before being immersed in cold fixative. Of the two types of axonal terminal identified in the nerves, one contained mainly large dense-cored vesicles, and the second contained numerous small vesicles. In specimens incubated in 5-hydroxydopamine, the small vesicles of the latter terminals exhibited the electron-dense cores which are characteristic of adrenergic axonal terminals. Counts made on perfused specimens showed that, in both the thoracic and cervical parts of the trachea, the density of adrenergic terminals was higher than that of terminals containing mainly large dense-cored vesicles. Overall terminal density was, however, higher in the thoracic than in the cervical part of the trachea, and estimates of nerve size showed that this was associated with the presence in the thoracic plexus of a substantially greater proportion of nerves with less than 6 axons. The possible function of the nerves in the control of the calibre of the submucous blood vessels was discussed. 相似文献
18.
Summary A preembedding dual immunolabeling technique and electron microscopy were utilized to demonstrate the localization of immunoreactive substance P and methionine-enkephalin-octapeptide (Enk-8) in ultrathin sections of the surface layer (laminae I and II) of rat spinal dorsal horn. The immunoreaction of Enk-8 was visualized as goldtoned silver particles and that of substance P as diaminobenzidine reaction products. Axonal terminals with immunoreactive substance P, and also unlabeled axonal terminals, formed synaptic junctions with the perikarya and dendritic processes of Enk-8-containing neurons. Dendritic profiles immunolabeled for substance P were synaptically linked with unlabeled axons but not with Enk-8-positive ones. Furthermore, it was found that Enk-8 axons and substance P axons terminated synaptically in juxtaposition to one another on the same immunonegative dendrites. Among the Enk-8-containing neurons axonal profiles also appeared to be synaptically associated with immunoreactive Enk-8 dendritic processes. 相似文献
19.
An electron microscope study of retrogradely labeled pyramidal neurons in layer VI of the primary auditory cortex (AI) after injecting horseradish peroxidase (HP) into the medial geniculate body was carried out in cats. Not less than 57.8±1.9% on average of the perimeter of perikaryon profiles of corticogeniculate neurons labeled with HP were found to be covered with astroglia processes. Between three and eight synapses occupying an average of 10.8±1.0% of the perimeter length were found on the perikaryon profiles of these neurons. Nearly all synapses (a total of 98.7%) at the soma of corticogeniculate neurons had symmetrical active zones, being made up of axonal terminals with flattened synaptic vesicles. Anterogradely HP-labeled axonal terminals of geniculocortical fibers were also found in the neuropil of layer VI in area AI, in addition to retrogradely labeled neurons. They contained large round synaptic vesicles and formed asymmetrical synapses. The potential role of axosomatic synapses in the shaping of corticogeniculate neuronal activity is discussed.A. A. Bogomolets Institute, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 2, pp. 171–178, March–April, 1990. 相似文献
20.
Immunocytochemistry of glutamate at the synaptic level 总被引:2,自引:0,他引:2
J Storm-Mathisen O P Ottersen 《The journal of histochemistry and cytochemistry》1990,38(12):1733-1743
High concentrations of glutaraldehyde (2-5%) were found optimal for fixation of glutamate. In the absence of glutaraldehyde, (para)formaldehyde does not permanently retain L-[3H]-glutamate or D-[3H]-aspartate previously taken up into brain slices. Rats were fixed by rapid transcardial perfusion with 2.5% glutaraldehyde/1% (para)formaldehyde, and brain samples osmicated, embedded in epoxy resin, sectioned, and exposed to specific antisera to glutamate (conjugated to carrier protein by glutaraldehyde), followed by colloidal gold-labeled second antibody. The gold particle density was higher over putative glutamatergic nerve terminals than over any other tissue elements (two to three times tissue average in cerebellum and hippocampus). Calibration by test conjugates containing known concentrations of fixed glutamate processed in the same fluid drops as the tissue sections indicated that the concentration of fixed glutamate in putative glutamatergic terminals in hippocampus CA1 was c. 20 mmol/liter. The grain density over the parent cell bodies was only slightly higher than the tissue average. (Grain densities over test conjugates of other amino acids, aldehyde-fixed to brain macromolecules, were similar to that over empty resin. Labeling was blocked by glutamate-glutaraldehyde but not by other glutaraldehyde-treated amino acids.) In other experiments, brain slices were incubated in oxygenated artificial cerebrospinal fluid (CSF) and then immersion-fixed and processed as above. Here, the ration of grain densities in putative glutamatergic terminals vs other tissue elements was greater than in perfusion-fixed material. Comparison of intra-terminal areas poor and rich in synaptic vesicles suggested that in this preparation vesicles contained at least three times the glutamate concentration of cytosol. In the glutamatergic synapses of the giant reticulospinal axons in lamprey the ratio was over 30. Prolonged K+ depolarization of hippocampal and cerebellar slices reduced the nerve terminal glutamate immunoreactivity in a Ca2(+)-dependent manner. The results suggest that glutamate is released by exocytosis at excitatory synapses and show that immunocytochemistry can be used to study the cellular processing of small molecules. 相似文献