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1.
The immunostaining pattern for the peptide gastrin/cholecystokinin 8 (gastrin/CCK8) in the molluscan central nervous system has been considered. The changes in the distribution of gastrin/CCK8 immunoreactivity were analyzed in the neurons of different areas of the cerebral ganglia (mesocerebrum and metacerebrum) and in the buccal ganglia of the terrestrial snail Helix aspersa, during rest and active phases. During the period of inactivity and after one day of activity, there were several immunoreactive neurons in the mesocerebrum and metacerebrum of the snails and in the buccal ganglia, whereas after 7 days of activity the number of labeled neurons decreased. Data suggested a storage of gastrin/CCK8 in the neurons when behavioral activities in which the peptide is involved (such as feeding-related behavior) are suppressed or reduced. The different percentage of gastrin/CCK8 immunoreactive neurons in the left and right mesocerebrum provides information about the activities controlled by these neurons, which could be related to the adaptive evolution and plasticity of the brain in terrestrial pulmonates.  相似文献   

2.
  • 1.1. FMRFamide immunoreactive neurons were detected in the central nervous system of the snail, Achatina fulica.
  • 2.2. FMRFamide immunoreactive neurons were found in all the ganglia comprising the central nervous system. In particular, the immunoreactivity was recognized in both the ordinary and giant neurons of the visceral and right parietal ganglia.
  • 3.3. In the cerebral and pleural ganglia, FMRFamide immunoreactive neurons were found only in the ordinary neurons. The immunoreactivity was shown to have a tendency to form a group in the cerebral and pedal ganglia.
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3.
Both immunoreactive intact cholecystokinin (CCK33) and its COOH-terminal octapeptide (CCK8) are detected in brain and gut extracts of monkey, dog, and pig using an antiserum with equivalent sensitivities for detecting CCK8 in the free form or when incorporated in the intact molecule. The failure to detect intact cholecystokinin in extracts from monkey or dog by using an antiserum developed by immunization with porcine CCK33 is due to marked species differences in the NH2-terminal portion of the molecule. Immunohistochemical staining reveals the presence of CCK peptides in rabbit cerebral cortical tissue neurons. Subcellular fractionation of rat cerebral cortical tissue demonstrates that CCK immunoreactivity is concentrated in the pellet identified by electron microscopy to contain a high proportion of synaptic vesicles. A converting enzyme that differs from trypsin has been partially purified from canine and porcine cerebral cortical extracts. It converts porcine CCK to smaller immunoreactive forms, but fails to convert big gastrin to heptadecapeptide gastrin. This enzyme differs from trypsin not only in substrate specificity but also in several physicochemical properties. Cerebral cortical extracts from hyperphagic ob/ob mice have strikingly lower contents of CCK than those from their lean littermates and other normal mice. These studies taken together are consistent with a role for CCK as a neurotransmitter involved in the overall regulation of appetite.  相似文献   

4.
Summary Gastrin/cholecystokinin (gastrin/CCK)-like immunoreactivity has been detected in the brain, suboesophageal ganglion and corpora cardiaca of the larva of Aeschna cyanea by radioimmunoassay and immunohistochemistry, by use of two antisera raised against the sulfated (CCK-8S) and the unsulfated form (CCK-8NS) of the carboxyl terminal octapeptide. Numerous immunoreactive neurons were demonstrated in the protocerebrum (exclusive of optic lobes) and suboesophageal ganglion where 20 and 15 symmetrical clusters of reactive cells, respectively, were observed. Immunoreactive cells also occurred in the tritocerebrum, the optic lobes and the frontal ganglion. In the corpora cardiaca, gastrin/CCK-like material was found both within intrinsic cells and axon terminals. RIA measurements support the immunohistochemical results in so far as large amounts of gastrin/CCK-like material were detected in the brain, corpora cardiaca and suboesophageal ganglion complex. Both boiling water-acetic acid- and methanol-extraction procedures were performed. Comparisons of the results lead to the conclusion that a large part of the gastrin/CCK-like material occurs as small molecules. Immunohistochemical procedures performed on material fixed in a solution of picric acid-paraformaldehyde demonstrated differences in the immunoreactivity of the tested antisera. First, the immunohistochemical reaction was always more pronounced when the CCK-8NS antiserum was used instead of the CCK-8S antiserum, which may be interpreted by a lower affinity of the latter. In the second place, some neurons strongly stained by the CCK-8NS antiserum were only very faintly if at all stained by the CCK-8S antiserum, which may mean that different peptides or at least distinct forms of the same precursor are detected.  相似文献   

5.
Summary The localization of the vertebrate-like neuropeptides substance P (SP), neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), and cholecystokinin (CCK8) in the central nervous system of the freshwater snail Planorbarius corneus has been studied using specific antisera and single and double immunohistochemistry. A widespread but precise distribution of immunore-activity (IR) in neurons and fibers of almost all the ganglia is observed for each antiserum. A comparison of the IR with classical neurosecretory staining (AB/AY) shows a partial overlap only for CGRP and CCK8. Whereas CGRP-IR is found in some Yellow Cells in the left parietal ganglion, CCK8-IR is found in Yellow Green, Green and Brown Cells in the viscero-parietal complex. Studies employing double-sequential methods or simultaneous immunofluorescence have shown that, with regard to the tested antisera, CCK8- and NPY-IR are colocalized in a limited number of cells and fibers in the buccal and visceral ganglia, whereas CCK8- and SP-IR are colocalized only rarely in neurons in the left cerebral ganglion. The possible roles in P. corneus of the investigated neuropeptides and the contribution that molluscan models may offer to the knowledge of the basic properties of neuropeptides are discussed.  相似文献   

6.
Carbohydrate polymers with acid?Cbase properties of extracellular matrix (ECM) have a critical impact on guidance, migration and synaptic transmission. The present study showed acid?Cbase properties of ECM stained with histochemical techniques. The central nervous system (CNS) of mature terrestrial slug Semperula maculata and freshwater bivalve Lamellidens corrianus was selected to visualize sulphated and carboxylic strong acid groups. Protein and carbohydrate molecules were found scattered in three different regions, viz. periganglionic sheath, along with the basement lamina, interperikaryal space including the perineuronal net, and neuropil, in both the molluscan species. Sulphated proteoglycans were abundant in periganglionic sheath of S. maculata and also occurred in the neuropil of L. corrianus. The interperikaryal space contained carboxyl groups along with hyaluronic acid, and neuropil contained abundant glycogen moiety in both species. Comparing the SDS?CPAGE of the neural tissue extracts revealed protein molecules that were found in the same fraction. The ECM of neuronal cells from S. maculata and L. corrianus presented different kinds of glycoproteins and proteoglycans, which showed its morphological as well as habitual differentiation.  相似文献   

7.
Cholecystokinin (CCK) activates the myenteric neurons of adult rats. The goal of this work is to determine the ontogeny of this activation by CCK-8 in the myenteric plexus of the duodenum (2 cm immediately following the pyloric sphincter aborally) and compare it with that of the dorsal vagal complex (DVC) - which occurs in 1-day old pups. Despite the existence of both of the CCK receptors, CCK1 and CCK2, in 4, 14, 21 and 35 day old rats, CCK-8 (0, 5, 10, 20 and 40 μg/kg, i.p.) increased Fos-like immunoreactivity (Fos-LI, a marker for neuronal activation) in the myenteric neurons of 21- and 35-day old rats but in the DVC of all age groups. As such, this belated activation of myenteric neurons by CCK-8 compared to the DVC may reflect a delayed role for these neurons in CCK-related functions.  相似文献   

8.
Prior studies have shown that the cerebral cortex cholecystokinin (CCK) receptor can bind CCK and gastrin analogs with high affinity. In the present work the brain CCK receptor had approximately a three times greater affinity for CCK8 than its C-terminal tetrapeptide (CCK4) while the C-terminal tripeptide (CCK3) was 1000-fold less potent than CCK4. Thus the C-terminal tetrapeptide appears to be the minimal C-terminal CCK sequence required for high affinity binding. Since brain membranes degrade various peptides including CCK, we also evaluated the stability of CCK analogs under the conditions used to measure receptor binding by the following three methods: (1) Studies of degradation-resistant analogs in binding assays; (2) analysis of analog degradation by high performance liquid chromatography (HPLC); and (3) determination of the change in potency of CCK analogs in competitive binding studies subsequent to preincubation with brain membranes. These studies indicated that degradation of analogs by the brain membranes although significant did not account for the differences in potency of analogs in competitive binding studies. Therefore, the observed differences in potencies of the analogs tested are due to the receptor affinity and not sensitivity of the analog to degradation.  相似文献   

9.
Summary The co-existence of immunoreactivities to substance P (SP), calcitonin gene-related peptide (CGRP), cholecystokinin (CCK) and dynorphin (DYN) in neurons of the dorsal root ganglion (DRG) of guinea-pigs has been investigated with a double-labelling immunofluorescence procedure. Four main populations of neurons could be identified that contained different combinations of these peptides and had distinctive peripheral projections: (1) Neurons that contained immunoreactivity to SP, CGRP, CCK and DYN were distributed mainly to the skin. (2) Neurons with immunoreactivity to SP, CGPR and CCK, but not DYN, were distributed mainly to the small blood vessels of skeletal muscles. (3) Neurons with immunoreactivity to SP, CGRP and DYN, but not CCK, were distributed mainly to pelvic viscera and airways. (4) Neurons containing immunoreactivity to SP and CGRP, but not CCK and DYN, were distributed mainly to the heart, systemic blood vessels, blood vessels of the abdominal viscera, airways and sympathetic ganglia. Other small populations of DRG neurons containing SP, CGRP or CCK alone also were detected. Perikarya containing these combinations of neuropeptides were not found in autonomic ganglia. The peripheral axons of neurons containing immunoreactivity to at least SP and CGRP were damaged by chronic treatment with capsaicin. However, some sensory neurons containing CCK alone were not affected morphologically by capsaicin.These results clearly show that individual DRG neurons can contain many different neuropeptides. Furthermore, the combination of neuropeptides found in any particular neuron is related to its peripheral projection.  相似文献   

10.
Endocrine cells containing gastrin/cholecystokinin (CCK)-like immunoreactivity were localized to the islet tissue in the pancreas of the spiny dogfish. Most of these cells were located in the 'intestinal' lobe of the pancreas; only occasional cells were observed in the 'splenic' lobe. The gastrin/CCK-like immunoreactive cells were often co-localized with the 'classical' pancreas hormones (insulin, glucagon and somatostatin). Radioimmunoassay of water extracts with a C-terminally directed antiserum revealed high levels of immunoreactive material in the intestinal part (48.6 +/- 19.9 pmol/g) and lower levels (4.5 +/- 0.6 pmol/g) in the splenic part. Acetic acid extracts of the intestinal lobe contained low levels (6.8 +/- 3.3 pmol/g) of gastrin/CCK-like immunoreactivity, whereas corresponding extracts of the splenic part showed no immunoreactivity. When the extracts were subjected to DEAE ion-exchange chromatography the gastrin/CCK-like peptides eluted as a major peak. After Sephadex gel filtration, pooled immunoreactive material from the main DEAE chromatographic peak eluted at a position close to that of CCK4. Further characterization by ion-exchange and reversed-phase HPLC showed that, in general, the immunoreactive material behaved like the shorter forms of the gastrin/CCK family (CCK4/G5 and CCK8/Cae 3-10).  相似文献   

11.
Enzyme histochemistry and immunocytochemistry were used to determine the distribution of neurons in the snail Helix aspersa which exhibited nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity and/or immunoreactivity to nitric oxide synthase (NOS). NADPH diaphorase-positive cells and fibres were distributed extensively throughout the central and peripheral nervous system. NADPH diaphorase-positive fibres were present in all neuropil regions of the central and peripheral ganglia, in the major interganglionic connectives and in peripheral nerve roots. NADPH diaphorase-positive cell bodies were found consistently in the eyes, the lips, the tentacular ganglia and the procerebral lobes of the cerebral ganglia; staining of cell bodies elsewhere in the nervous system was capricious. The distribution of NOS-like immunoreactivity differed markedly from that of NADPH diaphorase activity. Small clusters of cells which exhibited NOS-like immunoreactivity were present in the cerebral and pedal ganglia; fibres which exhibited NOS-like immunoreactivity were present in restricted regions of the neuropil of the central ganglia. The disjunct distributions of NADPH diaphorase activity and NOS-like immunoreactivity in the neurvous system of Helix suggest that the properties of neuronal NOS in molluscs may differ sigificantly from those described previously for vertebrate animals.  相似文献   

12.
The gastrodermal cell types of Microstomum lineare (Turbellaria, Macrostomida) were studied by electron microscopy. Their immunoreactivity (IR) to bovine pancreatic peptide (BPP), FMRF-amide and vasotocin, somatostatin, neurotensin, ACTH, CCK, bombesin, secretin, gastrin/CCK and insulin antisera was tested by light microscopic immunocytochemical methods. In addition to granular club cells and phagocytic cells, neurons and neoblasts occur in the gastroderm of this turbellarian species. This is the first observation of neurons in the gastroderm of a flatworm. Dense-core vesicles (70–100 nm diameter), electron lucent cytoplasm and numerous Golgi complexes characterize the neurons. Unpolarized two-way synapses, neuromuscular junctions and polarized chemical synapses can be observed in the gastroderm. Neoblasts with large nuclei and scanty cytoplasm and differentiating cells containing clusters of basal bodies occur next to the basal lamina of the gastroderm. BPP-like, FMRF-amide-like and vasotocin-like immunoreactivity is demonstrated in the gastroderm. Both BPP and FMRF-amide IR is restricted to the basal cytoplasm of the granular club cells, while a different location for IR to vasotocin antiserum is observed. The status of the neuronal cell in the gastroderm of M. lineare is discussed in relation to endocrine (paracrine) cells and neurons in the gastroderm of invertebrates.  相似文献   

13.
Summary The peptides cholecystokinin (CCK), neuropeptide Y (NPY), somatostatin (SOM), substance P (SP) and vasoactive intestinal peptide (VIP), and the synthesizing enzyme for acetylcholine, choline acetyltransferase (ChAT) were localized immunohistochemically in nerve cell bodies of the submucous ganglia in the small intestine of the guinea-pig. VIP-like immunoreactivity was found in 45% of submucous neurons. ChAT immunoreactivity was observed in a separate group of nerve cells, which made up 54% of the total population. There were three subsets of neurons immunoreactive for ChAT: (1) ChAT neurons that also contained immunoreactivity for each of the peptides CCK, SOM and NPY, representing 29% of all submucous neurons; (2) ChAT neurons that also contained SP-like immunoreactivity, representing 11% of all submucous neurons, and (3) ChAT cells that did not contain any detectable amount of the peptides that were localized in this study.  相似文献   

14.
We compared the mechanism of action of micronuclei (MN), unstable chromosome aberrations, and 8-hydroxydeoxyguanosine (8-OHdG) levels to evaluate the genotoxicity of methyl mercuric chloride (CH3HgCl) and mercuric chloride (HgCl2) in human peripheral lymphocytes. The chromosome aberrations in human peripheral lymphocytes exposed to various concentrations of CH3HgCl or HgCl2 increased in a concentration-dependent manner and were significantly higher than the control when the cells were incubated with 1 × 10−5 M (HgCl2) or 2 × 10−6 M (CH3HgCl). The increase in the incidence of micronucleated lymphocytes was significant among the exposed groups, being 2 × 10−5 M (HgCl2) and 5 × 10−6 M (CH3HgCl) compared with the control. CH3HgCl was about 4-fold more potent than HgCl2. We determined the 8-OHdG levels in human peripheral blood mononuclear cells(PBMC) and found that they were significantly higher in the exposed groups at 1 × 10−5 M (HgCl2) and 5 × 10−6 M (CH3HgCl) compared with the control. A detectable (p < 0.05) increase in the level of 8-OHdG was induced by CH3HgCl at a concentration that was about 50% of the amount of HgCl2 required to produce a similar response. The data confirmed the value of the MN and/or chromosome aberration assays for assessing of HgCl2- and/or CH3HgCl-induced genotoxicity, and indicated that they are about the same concentration as the 8-OHdG assay. The presence of genotoxic effects in peripheral blood lymphocytes exposed to the mercuric compounds indicated by the chromosome aberrations and the MN assays could be partly due either to the disturbance of the spindle mechanism, or to the elevated level of 8-OHdG brought by the generation of reactive oxygen species.  相似文献   

15.
Summary Using immunocytochemistry, gastrin/CCK-like immunoreactivity is found in sensory nerve cells in the ectoderm of the mouth region of hydra and in nerve cells in the endoderm of all body regions of the sea anemone tealia. These results are corroborated by radioimmunoassay: One hydra contains at least 5 fmole and one tealia at least 2 nmole gastrin/CCK-like immunoreactivity. Reactivities towards gastrin and CCK antisera with different specificities suggest that the coelenterate gastrin/CCK-like peptide contains the C-terminal amino-acid sequence common to mammalian gastrin and CCK. In addition the radioimmunochemical data indicate that the coelenterate peptide also contains an amino-acid sequence that resembles the sequence 20–30 of porcine CCK-33, but that no other sequences of gastrin are present. Thus, it is probably more CCK-like than gastrin-like.  相似文献   

16.
The distribution of neurones immunoreactive to antisera raised against the undecapeptide C-terminal fragment of drosulfakinin II (DrmSKII), Asp-Gln-Phe-Asp-Asp-Tyr(SO3H)-Gly-His-Met-Arg-Phe-NH2, has been studied in the blowfly Calliphora vomitoria. Antisera were preabsorbed with combinations of the parent antigen, the tetrapeptide Phe-Met-Arg-Phe-NH2 and cholecystokinin, the vertebrate sulfated octapeptide (CCK-8), Asp-Tyr(SO3H)-Met-Gly-Trp-Met-Asp-Phe-NH2, in order to ensure specificity for the sulfakinin peptides of C. vomitoria (the nonapeptide callisulfakinin I is identical to drosulfakinin I and callisulfakinin II differs from DrmSK II only by the presence of -Glu3-Glu4- in place of -Asp3-Asp4-). Only four pairs of sulfakinin-immunoreactive neurones have been visualised in the entire nervous system. These occur in the brain: two pairs of cells situated medially in the caudo-dorsal region close to the roots of the ocellar nerve and two other pairs at the same level but positioned more laterally. Despite the small number of sulfakinin-immunoreactive cells, there are extensive projections to many areas of neuropile in the brain and the thoracic ganglion. The pathway of the medial sulfakinin cells extends into each of the three thoracic ganglia and a metameric arrangement of sulfakinin neuronal projections is also seen in the abdominal ganglia. Neither the dorsal neural sheath of the thoracic ganglion, nor the abdominal nerves contain sulfakinin-immunoreactive material. These observations suggest that the sulfakinins of the blowfly function as neurotransmitters or neuromodulators. They do not appear to have a direct role in gut physiology, as has been shown by in vitro bioassays for the sulfakinins of orthopterans and blattodeans. In addition to the neurones that display specific sulfakinin immunoreactivity, other cells within the brain and thoracic ganglion are immunoreactive to cholecystokinin/gastrin antisera. There are, therefore, at least two types of dipteran neuropeptides with amino acid sequences that are similar to the vertebrate molecules cholecystokinin and gastrin.  相似文献   

17.
In the gastrin and/or cholecystokinin-like immunoreactivity (G/CCK-LI) elution patterns of blood cells in human adults, erythrocyte (RBC) elution pattern has three peaks which are coeluted with gastrin-34 (G34), gastrin-17 (G17) and Vt, and polymorphonuclear leukocyte (PMN) and mononuclear cell (MNC) elution patterns have four peaks which are coeluted with Vo, G34, G17 and Vt. The content of G/CCK-LI in RBC is 1.20±0.54 fmole/108 cells (means±SD). Than in PMN and MNC is 1.44±0.67 p mole/108 cells and 1.67±0.76 p mole/108 cells, respectively.  相似文献   

18.
Dibutyryl cyclic GMP has been reported to interact with antisera specific for C-terminal tetrapeptide amide common for cholecystokinin (CCK) and gastrin. Moreover, cyclic nucleotides elute by gel chromatography in the same position as the free CCK/gastrin tetrapeptide. Therefore, we have examined the reactivity of 25 mononucleotides with eight CCK and gastrin antisera. The results show that the nucleotides all bind poorly to the antisera (nucleotide concentration required 1 mM). Hence, endogenous cyclic nucleotides, which are present in biological extracts in pM to nM concentrations, do not interfere with immunochemical CCK or gastrin measurements. The antisera displayed highly individual patterns of reactivity without preferential binding of di- or monobutyryl cyclic nucleotides (AMP, GMP or IMP). Thus, the present results do not support the idea of structural resemblance between the C-terminus of CCK/gastrin peptides and butyryl derivatives of cyclic GMP. Enzymatic treatment of the antral tetrapeptide-like immunoreactivity showed that nucleotides do not contribute to this material, which appears exclusively peptidergic.  相似文献   

19.
The presence of high concentrations of both dopamine and cholecystokinin (CCK) in the striatum and in various limbic structures suggests that the CCK may not only influence dopaminergic transmission, but it also may be relevant to the psychopathology of schizophrenia and to the therapeutic effects of neuroleptics. By using a synaptosomal fraction isolated from the mouse cerebral cortex and [propionyl-3H]CCK8-sulphate ([3H]CCK8S) as a ligand, a single binding site for [3H]CCK8 with aK d value of 1.04 nM and aB max value of 42.9 fmol/mg protein was identified. The competitive inhibition of [3H]CCK8S binding by related peptides produced an order of potency of CCK8-sulphated (IC50=5.4 nM)>CCK8-unsulfated (IC50=40 nM) and >CCK4 (IC50=125 nM). The regional distribution of [3H]CCK8S binding in the mouse brain was highest in the olfactory bulb (34.3±5.6 fmol/mg protein) > cerebral cortex > cerebellum > olfactory tubercle > striatum > pons-medulla > mid brain > hippocampus > hypothalamus (12.4±2.1 fmol/mg protein). The repeated administration of haloperidol (2.5 mg/kg/tid) increased the binding of [3H]CCK8S in cerebral cortex from 31.8±1.7 to 38.9±5.2 fmol/mg protein. The varied distribution of CCK8S receptors may signify nonuniform functions for the octapeptide in the brain.  相似文献   

20.
To elucidate the role of aquaporins in the control of the root pressure, we tested the effects of HgCl2 (aquaporin blocker) at concentrations from 10?8 to 10?2 M on the exudation rate (J w). Experiments were performed with detached roots of 5–7-day-old etiolated maize (Zea mays L.) seedlings. HgCl2 suppressed exudation by 50–70% at the concentration of 2 × 10?5 M. At the concentration of 2.5 × 10?4 M, HgCl2 reduced J w during first 20–40 min, but in 2 h, it activated exudation by ten and more times. In this case, the root and osmotic pressures of the exudates increased by 1.5 times. The hydraulic conductance reduced approximately by 30% during first 30 min and increased severalfold in 2 h. The temperature coefficient (Q10) of J w attained 14 in 2 h. At the concentration of 10?2 M, HgCl2-induced acceleration of exudation was replaced by its inhibition essentially immediately. We suggested that a driving force for HgCl2-induced stimulation of the J w might be an increase in the osmotic component of the root pressure or the involvement of its nonosmotic (so-called metabolic) component. The results allow a supposition that aquaporins are involved in the control of water transport in the root.  相似文献   

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