共查询到20条相似文献,搜索用时 15 毫秒
1.
Yaffa Mizrachi Peter I. Lelkes Richard L. Ornberg Gertrude Goping Harvey B. Pollard 《Cell and tissue research》1989,256(2):365-372
Summary Chromaffin cells in the adrenal medulla are found in close proximity to capillary endothelial cells, thereby forming the classical endocrine complex. To examine the possible chemical basis of their interaction in more detail, we have grown bovine adrenal medullary endothelial (BAME) cells in monolayer cultures and added to them pheochromocytoma (PC12) cells, a chromaffin tumor cell line of rats. The PC12 cells were chosen because of the similarities they share with adrenal medullary chromaffin cells. PC12 cells rapidly attached to BAME cells cultures, their rate of adhesion being significantly enhanced over binding of PC12 cells to either uncoated plates or to monolayers of unrelated cell cultures. Consistent with this observation, we noted that the extracellular matrix (ECM) derived from the BAME cells did not enhance PC12 cell adhesion and did not promote neurite sprouting as previously described for ECM derived from corneal endothelial cells. The specific adhesion between PC12 and BAME cells could be abolished by cell surface extracts derived from these two cells but not by extracts derived from unrelated cell types. This activity was heat-labile, sensitive to trypsin and, to a lesser extent, to neuraminidase. We therefore conclude that PC12 cells may interact with BAME cells by specific proteinaceous adhesive factors associated with their plasma membranes. These interactions might represent the formative role of cell-cell contacts in the organization of the developing adrenal gland.Abbreviations
BAME
bovine adrenal medullary endothelial cells
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DMEM
Dulbecco's modified essential medium
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ECM
extracellular matrix
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EMEM
Eagle's modified essential medium
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FCS
fetal calf serum
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PBS
phosphate-buffered saline
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PC12
rat pheochromocytoma cells 相似文献
2.
Summary An autoradiographic study was performed on the effects of hydrocortisone, reserpine, propranolol and phentolamine on the uptake of tritiated amines by adrenal medullary cells of the mouse. Oral feeding of hydrocortisone had no significant effect on the normal uptake pattern of dopamine, noradrenaline or adrenaline by medullary cells of different type (A cells or NA cells) or location (marginal or central), although the overall amounts taken up were markedly reduced. Handling the animals led to similar reductions in the uptake of all three amines and was thus clearly shown to be the important factor in this effect. Reserpine reduced the uptake of [3H] noradrenaline to 25 % of the control value although the relative distribution remained unchanged. Propranolol and phentolamine had no observed effect on [3H] noradrenaline uptake. These results are discussed in the light of the previously reported action of ACTH in reversing the effects of hypophysectomy on medullary amine uptake (Hirano and Kobayashi 1978), and it is concluded that ACTH must exert this effect directly on the adrenal medulla rather than through the secretion of adrenal corticosteroids. It is also suggested that reserpine acts, as in neurons, by blocking amine uptake into intracellular granules rather than by blocking uptake into the cell itself. 相似文献
3.
Summary Studies on isolated adrenal chromaffin cells in primary cultures may be seriously hampered by the presence of non-chromaffin, mainly fibroblast-like cells, which always occur in dissociates of adrenal medullary tissue and often outnumber the chromaffin cells by the end of the first week of culture, when no measures are taken to control their proliferation. The present study offers a new means to inhibit effectively the proliferation of these accessory cells by treating the cultures with dibutyrylic cyclic AMP (dbcAMP, 0.1 or 0.01 mM) and equimolar amounts of the phosphodiesterase inhibitor theophylline. With this treatment cultures of young rat adrenal chromaffin cells remain virtually free of accessory cells for two weeks of culture. Cultures of bovine adrenomedullary cells retain their initial amounts of non-chromaffin cells, which largely depends upon whether the primary cell suspensions have undergone differential plating prior to seeding. Suppression of accessory cell proliferation with dbcAMP and theophylline is partly due to maintaining differentiation of cortical cells, which otherwise dedifferentiate into rapidly dividing fibroblast-like elements. However, a more direct action of dbcAMP on accessory cells in terms of growth control is also conceivable. DbcAMP and theophylline in the doses applied do not impair the viability, ultrastructure and catecholamine-storing capacity of cultured chromaffin cells. 相似文献
4.
Summary A method of cultivation involving both repeated trypsinisations (at room temperature) and explantation of the tissue fragments on polythene discs has been shown to be apt to the growth in vitro of rat adult decapsulated adreno-cortical tissue. This is the first time that the successful cultivation of such a tissue is reported. The technique and its applications are discussed.The effects of 1–24 corticotrophin (ACTH1–24) on the rat adult adrenal cultures have been examined by both electron microscopy and autoradiography. Zona fasciculata and reticularis cells grown in the absence of ACTH for long terms (15–16 days) survive and proliferate as dedifferentiated elements. If ACTH1–24 is added to the cultures, adrenocortical cells will, within 2 days, simultaneously increase their proliferation rate and differentiate. After 7 days of treatment, cortical cells exhibit not only fully differentiated but even hypertrophic morphologic features. Significant stimulations of adrenal DNA, RNA and gross protein synthesis have been found to take place at different times after the starting of the ACTH1–24 treatment. These data are discussed in relation to the findings previously reported in literature.Rat adult adrenal gland tissue cultures are proposed as a non-previously available tool for investigations into the physiopathology of the adrenal cells to be carried out in a carefully controlled environment.A Preliminary report on part of this material was given at the annual meeting of the Société Française de Microscopie Elécronique, Nantes, May 1972.Authors wish to thank Drs. P. G. Andreis and A. S. Belloni for their skilful assistance in the autoradiographic experiments. Thanks are also due to Mr. G. Gottardo for his excellent technical help in electron microscopic work. This work was partly supported by a contract with the CNR-Italy (No 69.0172/115.3439). 相似文献
5.
Summary Transplantation of a small fragment of the ileum beneath the kidney capsule in syngenic mice results in the formation of a cyst lined with proliferating intestinal epithelium. The duration of the cell cycle in this epithelium was determined (using tritiated thymidine and the FLM method) as 14.5 h, as compared with 11.5 h in the intestinal epithelium in situ. We conclude that the intestinal content has little effect on the cell cycle of epithelial cells of the small intestine. 相似文献
6.
Katsuko Kataoka 《Cell and tissue research》1970,103(2):170-178
Summary The duodenal and colonic epithelia in mice were observed with electron microscopic autoradiography 2, 5 and 24 hours after a single injection of 3H-thymidine. After 2 hours, in the duodenum, silver grains are found in many undifferentiated cells, in a few young goblet cells, in some crystal-containing cells, and in some lymphocytes. In the colon after 2 hours silver grains are seen in some undifferentiated cells, and in many young goblet cells. Undifferentiated cells are characterized by a few short microvilli, poorly developed rough-surfaced endoplasmic reticulum, abundant free ribosomes, and a few apical moderately dense granules. In normal animals, absorptive cells seem to arise from undifferentiated cells, and goblet cells — from younger goblet cells. Undifferentiated cells could also become young goblet cells. Crystal-containing cells, which may not be of epithelial origin, proliferate in the epithelium in the adult animal. 相似文献
7.
Summary The adrenal medullae of rats were studied electron microscopically from day 13.5 of gestation. Synapses with thickening of pre- and post-synaptic membranes were first evidenced on the medullary cells of 15.5-day-old fetuses. They increased gradually in number with advancing age. In the medullary cells, a few membrane-limited granules were recognized on day 13.5 of gestation, and thereafter they increased in number. The appearance of the granules was not uniform; some of the granular contents consisted of fine or coarse particles and others contained homogeneous material of varying electron-densities. The population of these granules in the cytoplasm was different from cell to cell. Thus, the discrimination of cell types (NA- and A-cells) was not possible in the prenatal period. Granular discharge of the cells was totally absent in the normal untreated fetuses. However, upon the intraperitoneal administration of insulin to the fetus on day 16.5, 18.5, or 21.5 of gestation, granular discharge by reverse pinocytosis was first evidenced in the medullary cells of the 21.5-dayold fetus. 相似文献
8.
Muscarine stimulated a concentration-dependent accumulation of [3H]inositol phosphates in bovine adrenal medullary cells preloaded with [3H]inositol. This muscarinic activation of inositol phospholipid metabolism was fully inhibited by the -ligand 1,3-di-o-tolylguanidine (DTG) with an IC50 of approximately 45 M. Higher concentrations (100 M) of (+) N-allylnormetazocine (SKF-10047) also partially inhibited this response. A concentration of DTG sufficient to fully inhibit the muscarinic response also produced a significant partial inhibition of [3H]inositol phosphate accumulation in response to histamine but not to angiotensin II. These data demonstrate that -compounds inhibit agonist-stimulated inositol phospholipid metabolism in bovine adrenal medullary cells, with a degree of selectivity towards the muscarinic response. 相似文献
9.
Summary In a total of 120 rats the number of 3H-Tdr-labelled nuclei of the adrenocortex was counted during various phases of the estrous cycle. In two separate experiments cyclic variations in the number of DNA synthesizing cells were found. The labelling index of the zona glomerulosa showed a significant maximum in proestrus whereas in zona fasciculata a minimum was revealed in estrus. In the glomerulosa the maximum of labelled cells was followed by a mitotic peak in estrus. Due to very low numbers of labelled cells no rhythm was found in the reticularis. The possible endocrine regulations of this endogenous rhythm of adrenocortical cell replication are discussed.The authors wish to express their appreciation for the expert assistance in the statistical evaluation of data rendered by Dr. Trieb and for the reliable technical aid by Mr. Majer 相似文献
10.
Summary Light-microscopic autoradiographs of the adrenal medulla at various intervals after the intravenous injection of [3H] 5-HTP, [3H] 5-HT, [3H] noradrenaline and [3H] adrenaline have been studied. The distribution of silver grains following [3H] 5-HTP uptake was found to be uniform over each of the two main cell populations, adrenaline-storing (A) cells and noradrenaline-storing (NA) cells in the adrenal medulla, but A cells were twice as active as NA cells in incorporating the isotope, a situation very similar to that found after [3H] dopa uptake. 5-HT administration resulted in a pattern resembling the distribution of [3H] noradrenaline uptake, with A cells being 4 or 5 times more active than NA cells and a gradient of activity from the periphery of the medulla inwards. However, the time-course for the loss of radioactivity was not the same for both amines: levels of 5-HT activity were not significantly reduced after one week whereas the degree of [3H] noradrenaline labelling after one week was less than 10% of that at one hour. Thus 5-HT may be bound to sites in the adrenal medulla normally occupied by noradrenaline but it would appear that the release mechanism is different. There was no evidence of 5-HT uptake by adrenal nerve endings. 相似文献
11.
Heidar Toolee Tayebeh Rastegar Somayeh Solhjoo Keywan Mortezaee Mahshid Mohammadipour Iraj Regerdi Kashani Mohammad Akbari 《Journal of cellular biochemistry》2019,120(4):5042-5054
Kisspeptin (Kp) expression in testis has caused most of the recent research surveying its functional role in this organ. This peptide influences spermatogenesis and sperm capacitation, so it is considered as a regulator of reproduction. Kp roles exert through hypothalamic/pituitary/gonadal axis. We aimed to evaluate direct roles for Kp on proliferation and differentiation of spermatogonial cells (SCs) when the cells are cocultured with somatic cells. Somatic cells and SCs were isolated from adult azoospermic and newborn mice and then enriched using a differential attachment technique. After the evaluation of identity and colonization for SCs, the cells were cocultured with somatic cells, and three doses of Kp (10−8-10−6 M) was assessed on proliferation (through evaluation of MVH and ID4 markers) and differentiation (via evaluation of c-Kit and SCP3, TP1, TP2, and, Prm1 markers) of the coculture system. Investigations were continued for four succeeding weeks. At the end of each level of testosterone in the culture media was also evaluated. We found positive influence from Kp on proliferative and differentiative markers in SCs cocultured with somatic cells. These effects were dose-dependent. There was no effect for Kp on testosterone level. From our findings, we simply conclude that Kp as a neuropeptide for influencing central part of reproductive axis could also positively affect peripheral processes related to spermatogenesis without having an effect on steroidogenesis. 相似文献
12.
Adrenal medullary chromaffin cells are derivatives of the neural crest and are widely believed to share a common sympathoadrenal (SA) progenitor with sympathetic neurons. For decades, the adrenal cortical environment was assumed to be essential for channelling SA progenitors towards an endocrine chromaffin cell fate. Our recent analysis of steroidogenic factor 1(Sf1) −/− mice, which lack an adrenal cortex, has challenged this view: in Sf1 −/− mice chromaffin cells migrate to the correct “adrenal” location and undergo largely normal differentiation. In contrast to Sf1 homozygous mutants, heterozygous animals have an adrenal cortex, which, however, is smaller than in wildtype littermates. We show here that the Sf1 +/− adrenal cortical anlagen attract normal numbers of chromaffin progenitor cells into their vicinity by embryonic day 13.5 (E13.5). Two days later, however, only a few scattered cells with highly immature features have immigrated into the adrenal cortex, whereas the remainder form a coherent cell assembly ectopically located at the medial surface of the gland. These cells appear more mature than the scattered intracortical chromaffin progenitors and express the adrenaline synthesizing enzyme PNMT with a delay of 1 day in comparison with wildtype littermates. Nevertheless, chromaffin progenitor cells undergo a numerical reduction of approximately 30% by E17.5. Together, our data suggest that normal adrenocortical development is critical for the correct immigration of chromaffin progenitors into the cortical anlagen, for the timing of PNMT expression and for the regulation of chromaffin cell numbers.This work was supported by a grant from the Deutsche Forschungsgemeinschaft (SFB 488, TP A6). 相似文献
13.
In newborn rats serotonin was found to bind the membranes of adrenal medullary cells, to the wall of their vesicles and could be demonstrated also in the nucleus. The observations allowed conclusions concerning the cellular mechanisms of serotonin action. 相似文献
14.
Alan K. Percy John F. Moore Gordon A. Plishker Jack C. Waymire 《Neurochemical research》1991,16(4):505-511
Cultured adrenal chromaffin cells, representing a virtually homogeneous population of neuronai elements, have been utilized to examine the final enzymes in the formation of phosphatidylcholine (PC) and phosphatidylethanolamine (PE), namely, choline phosphotransferase, ethanolaminephosphotransferase, and the N-methyltransferases in the sequential methylation of PE to PC. Each enzyme has been characterized extensively in terms of substrate requirements, pH optima, detergent and cation effects, and response to inhibitors revealing properties very similar to those in other neural preparations. The respective activities are stable for up to two weeks of adrenal chromaffin cell culture suggesting that this system is a suitable model for examining the relative roles and the regulation of each pathway in PC formation.Abbreviations EPT
ethanolaminephosphotransferase
- CPT
cholinephosphotransferase
- NMT
N-methyltransferase
This work supported by funds provided to the Section of Pediatric Neurology by Texas Children's Hospital. 相似文献
15.
Ghanim Almahbobi 《Cell and tissue research》1995,281(2):387-390
High-resolution field emission scanning electron microscopy was used to study the organisation of intermediate filaments around lipid droplets and their binding to these droplets, in primary culture of bovine adrenal cells. Whole-mount preparations of intermediate filaments and bound lipid droplets were prepared from cells grown on Formvar-coated grids and processed by freeze-drying. Intermediate filaments were seen as an interconnected network enveloping the entire droplet. The bound filaments appear to be directly adherent to the surface of the droplet and hence take on its curved contour. The binding of the filaments to the droplets was determined by means of tilting. This study provides a new approach to investigate the cytoskeleton and its associated structures with high-resolution three-dimensional images. 相似文献
16.
Jun-Tao Feng Xiu-Ming Wu Xiao-Zhao Li Ye-Qiang Zou Ling Qin Cheng-Ping Hu 《Respiratory research》2012,13(1):1-9
Background
Studies have shown that epinephrine release is impaired in patients with asthma. The pregnancy of female rats (dams) with asthma promotes in their pups the differentiation of adrenal medulla chromaffin cells (AMCCs) into sympathetic neurons, mediated by nerve growth factor, which leads to a reduction in epinephrine secretion. However, the relatedness between the alteration of AMCCs and increased asthma susceptibility in such offspring has not been established.Methods
In this study, we observed the effects of allergization via ovalbumin on rat pups born of asthmatic dams.Results
Compared to the offspring of untreated controls, bronchial hyperreactivity and airway inflammation were more severe in the pups from sensitized (asthmatic) dams. In pups exposed to nerve growth factor (NGF) in utero these effects were aggravated further, but the effects were blocked in pups whose dams had been treated with anti-NGF. Furthermore, alterations in AMCC phenotype corresponded to the degree of bronchial hyperreactivity and lung lesions of the different treatment groups. Such AMCC alterations included degranulation of chromaffin granules, reduction of epinephrine and phenylethanolamine-n-methyl transferase, and elevation of NGF and peripherin levels.Conclusions
Our results present evidence that asthma during the pregnancy of rat dams promotes asthma susceptibility in their offspring, and that the transformation of AMCCs to neurons induced by NGF plays an important role in this process. 相似文献17.
Dr. Gastone Giovanni Nussdorfer 《Cell and tissue research》1970,103(3):398-409
Summary The structural changes of the zona juxtamedullaris of the rat adrenal cortex at birth, have been examined by the light and the electron microscope. In this zone clusters of medullary cells lying among the strands of cortical tissue were observed. In the inner portion of the zona juxtamedullaris two types of adrenocortical cells were found: light and very-dark cells. The latter are smaller than the light cells and are always in close connection with the medullary tissue. The ultrastructural features of the very-dark cells suggest that these elements are in degeneration. This finding supports the hypothesis that at birth there is a partial degeneration of the rat zona juxtamedullaris, i.e. the zone corresponding to the fetal zone of some mammalian species.It is proposed that in all mammalian species at birth there is a partial regression of the zona juxtamedullaris and that the regression of the fetal zone is only the quantitative increase of this phenomenon. This hypothesis is discussed in relation to numerous data demonstrating that there are enzymatic conditions in the rat during fetal life, which permit a discrete hypertrophy of the adrenal cortex.The author wishes to express his appreciation to Dr. A. Gambino and to Mr. G. Gottardo for technical assistance. 相似文献
18.
Summary Expiants of adrenal medullary tissue taken from newborn guinea pigs were grown in culture for up to two weeks. The explants exhibited sparse outgrowth of neurite-like processes, in contrast to adrenal medullae taken from young postnatal rats or adult guinea pigs that were (i) grown under identical conditions (Unsicker and Chamley 1977) or (ii) transplanted to the anterior chamber of the eye (Unsicker et al. 1981), respectively. Nerve growth factor (10–100 ng/ml, 2.5S NGF) did not enhance formation of processes. However, electron-microscopic investigations revealed the presence of numerous processes within the explants, which extended from chromaffin cells and were characterized by longitudinally oriented cytoskeletal structures, various populations of clear and dense-cored vesicles, varicosities and growth cones. Chromaffin cell bodies largely resembled their in situ-counterparts, but had fewer and smaller storage vesicles than controls.The results are discussed in light of recent findings regarding the potency of NGF and NGF-like growth factors to induce neuronal transdifferentiation of adrenal chromaffin cells.Supported by grants from the Deutsche Forschungsgemeinschaft (SFB 103 and Un 34/6) 相似文献
19.
Dr. Carl E. Creutz 《Cell and tissue research》1977,178(1):17-38
Summary Myosin was isolated in high purity from the bovine adrenal medulla by gel filtration and ion exchange chromatography. The purified myosin was analyzed by electrophoresis in gels containing SDS and found to contain a 200,000 molecular weight heavy chain and major light chains of molecular weights 20,000 and 17,000 in a 111 molar ratio. At high ionic strength the myosin had high Ca-ATPase and K-EDTA-ATPase activities and low Mg-ATPase activity. At low ionic strength, the Mg-ATPase was activated to a low level by rabbit muscle actin. The myosin was found to decorate F-actin in the absence, but not the presence of ATP. In low ionic strength solutions, the myosin assembled into characteristic bipolar filaments.The distribution of this myosin in the adrenal medulla and of cross-reacting myosin in several other bovine tissues was determined with the use of antimedullary myosin immunoglobulin G as a specific stain that was detected by direct and indirect immunofluorescence. In the medulla strong staining was seen between the chords of chromaffin cells indicating the presence of a highly muscular vasculature that may perform functions analogous to those of the myoepithelium of exocrine glands. The chromaffin cells showed weak positive staining around the nuclei and in a pattern radiating toward adjacent blood vessels. Cells of the inner zone of the adrenal cortex showed strong staining in the peripheral cytoplasm while cells in the intermediate and outer zones did not stain. In a blood smear, platelets and the cytoplasm of leukocytes stained strongly while erythrocytes did not stain. In striated muscle and the gray and white matter of the cerebrum only the capillaries and larger vessels stained. In the liver the phagocytic cells bordering vascular sinuses stained strongly while the hepatocytes were separated from one another by a 2 micron trilaminar band possibly representing the microfilament web surrounding the bile canaliculi and associated with junctional complexes.The results suggest that myosin is present in several highly differentiated, non-motile tissue cells where it may play a role in secretion or other specialized functions.The author gratefully acknowledges the support and encouragement received from Francis D. Carlson (Johns Hopkins University) and Harvey B. Pollard (National Institutes of Health) in whose laboratories the majority of this work was performed, as well as additional advice and assistance from John Cebra, Richard Cone, William F. Harrington, Shin Lin, Robert Wyllie and the members of their laboratories 相似文献
20.
目的:原代培养新生小牛肾上腺髓质嗜铬细胞,观察离体条件下肾上腺髓质细胞的生长和儿茶酚胺的分泌特性。方法:密度梯度离心和差速贴壁法分离和纯化肾上腺髓质细胞,ELISA检测培养上清液中肾上腺素和去甲肾上腺素的浓度变化。结果:肾上腺髓质嗜铬细胞生长良好。4~6周起细胞出现终极分化。培养液中儿茶酚胺浓度在第2~8 d内稳定,第10 d开始明显下降。结论:应用单细胞培养法可以成功建立原代肾上腺髓质嗜铬细胞,并可在原代培养前8d内进行细胞行为学的研究。 相似文献