Elektronenmikroskopische Untersuchungen am Nebennierenmark von Maus, Meerschweinchen und Katze

Wetzstein, R.

doi:10.1007/bf00339807

Cited by 103 publications

(15 citation statements)

References 45 publications

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“…The methods used 478 RUTH V.BAKER in the present work do not give an answer to this problem. However, in view of the observations on the chromaffin granules of the adrenal medulla (Lever, 1955;Sj6strand & Wetzstein, 1956;Wetzstein, 1957) and on mast cell granules (Smith & Lewis, 1957), which have revealed the presence of both mitochondria and a second kind of granule, it seems likely that all types of amine-carrying granules are intracellular structures distinct from mitochondria.…”

Section: Ruhvbaementioning

confidence: 99%

Mitochondria and storage granules for 5‐hydroxytryptamine

Baker¹

1959

The Journal of Physiology

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A large fraction of the total 5-hydroxytryptamine (5-HT) in a homogenate of the dog's duodenal mucosa, prepared in isotonic sucrose solution, is present in an intracellular structure that is sedimented in the gravitational field used for the sedimentation of mitochondria (Baker, 1958a).The intracellular localization of 5-HT presents many analogies to the storage of both histamine and the catechol amines. These amines are also present in granules, and it has recently been shown that the amine-carrying granules are distinct from mitochondria, although both are sedimented together in isotonic sucrose (Blaschko, Hagen & Hagen, 1957; Barrnett, Hagen & Lee, 1958). This separation was effected by the use of the density gradient method of centrifugation. This method has now been applied to granules obtained from the dog's duodenal mucosa in order to obtain evidence on the localization of 5-HT in relation to the mitochondrial enzymes. Some of these results have already been briefly reported (Baker, 1958b). METHODSPreparation of the large-granule fraction. Homogenates of the dog's duodenal mucosa were prepared as previously described (Baker, 1958a), and centrifuged at low speed for 20 min at 900 g. This low-81eed supernatant was decanted and centrifuged for 30 min at 11,000 g to give a high8peed supernatant and a high-speed sediment. The high-speed sediment was resuspended in 0-3m sucrose to a final volume of 10 ml. This suspension is referred to as the large-granule fraction. Ultracentrifugations were carried out in the SPINCO ultracentrifuge, using the swinging bucket rotor SW 39L.Preparation of the density gradient. The gradient was prepared in the cellulose ultracentrifuge tubes by layering measured volumes of sucrose solutions of different concentrations one above the other in order of decreasing molarity. The gradient tubes were prepared 18 hr before use and kept in a refrigerator at 4V C. Immediately before centrifugation 1 ml. of the large-granule fraction was placed in each tube above the gradient and the tubes were centrifuged at 103,000 g for 1 hr. The positions of the opaque layers after centrifugation were noted and the tubes were cut using the cutter designed by Dr E. H. J. Schuster. This cutter is similar to the one described by Randolph & Ryan (1950). The volume of each fraction was noted and determinations were carried out immediately.

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Section: Ruhvbaementioning

confidence: 99%

Mitochondria and storage granules for 5‐hydroxytryptamine

Baker¹

1959

The Journal of Physiology

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“…This has extended rather than limited the range of possible mechanisms. It is clear, however, from the cell fractionation studies initiated by Blaschko & Welch (1953) and by Hillarp, Lagerstedt & Nilson (1953) and from electronmicroscopical evidence provided by Hillarp et al (1953), Lever (1955), Wetzstein (1957), De Robertis & Vaz Ferreira (1957), and many others, that most of the catecholamines in the chromaffin cell are present in membrane-limited subcellular structures ranging in size from about 500 to 2,000 A and known as chromaffin granules (see reviews by Weiner, 1964;Coupland, 1965a;Vane, 1959). But not all medullary catecholamines are recovered in chromaffin granules when the cells are disrupted.…”

Section: The Adrenal Chromaffin Cellmentioning

confidence: 99%

“…Microscopists prefer to believe that the chromaffin granules are more immediately involved and offer a variety of suggestions, supported by appropriate micrographs, about how release occurs. These suggestions range from leakage of amines from granules to cytoplasm and thence, presumably by diffusion, across the plasmalemma to the cell exterior (for example, Wetzstein, 1957;Hillarp, Hokfelt & Nilson, 1954;De Robertis & Vaz Ferreira, 1957) to expulsion of intact granules through the cell membrane (for example, Cramer, 1928;Smitten, 1965). One of the more obvious limitations of the electronmicroscopical approaches (apart from the impossibility of assessing the involvement of " free" amines) is that while a given image may suggest a means whereby the cell could release its contents, the image cannot answer the question whether this is the mechanism of release, or even an important one: the quantitative evidence and correlation with secretory state that would justify such a conclusion would be difficult to obtain and is certainly not available.…”

Section: The Adrenal Chromaffin Cellmentioning

confidence: 99%

Stimulus‐secretion coupling: the concept and clues from chromaffin and other cells

Douglas

1968

British J Pharmacology

1,365

414

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“…On the other hand, it has frequently been suggested, principally on evidence obtained with light or electron microscopy, that the chromaffin granules may be more directly involved and transfer their amines, in one way or another, to the cell exterior during the secretory response (e.g. Cramer, 1928;Hillarp, Hokfelt & Nilson, 1954;De Robertis & Vas Ferreira, 1957;Wetzstein, 1957Wetzstein, , 1961De Robertis & Sabatini, 1960;Coupland, 1965). These granules, according to Hillarp (1960b), are of two types: the classical 'heavy ' chromaffin granule in which catecholamine is stored in equivalent amounts with adenine nucleotides (principally ATP), and a less numerous 'light' chromaffin granule in which the catecholamines are held without such nucleotide.…”

Section: Introductionmentioning

confidence: 99%

Evidence that the secreting adrenal chromaffin cell releases catecholamines directly from ATP‐rich granules

Douglas¹,

Poisner²

1966

The Journal of Physiology

112

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SUMMARY1. Cats' adrenal glands were perfused with Locke's solution and stimulated through the splanchnic nerves or by acetylcholine.2. In response to such stimulation there appeared in the venous effluent, in addition to catecholamines, large amounts of AMP and adenosine and smaller amounts of ATP and ADP. Like the catecholamines, these substances had their origin in the chromaffin cells as was shown by their failure to appear when the splanchnic nerves were stimulated during perfusion with drugs blocking the adrenal synapses.3. During stimulation the ratio of catecholamines: ATP and metabolites in the venous effluent corresponded closely with the reported ratio of catecholamines: adenine nucleotides in the 'heavy' chromaffin granules.4. Adenine nucleotide appeared in the adrenal effluent pari passu with catecholamines within a second or two of beginning stimulation.5. It is concluded that the nucleotide-rich granules are the immediate source of catecholamines released from the stimulated adrenal chromaffin cell, and that the other two intracellular 'pools' that have been described, nucleotide-poor and 'free' cytoplasmic catecholamines, contribute little or not at all.

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Elektronenmikroskopische Untersuchungen am Nebennierenmark von Maus, Meerschweinchen und Katze

Cited by 103 publications

References 45 publications

Mitochondria and storage granules for 5‐hydroxytryptamine

Mitochondria and storage granules for 5‐hydroxytryptamine

Stimulus‐secretion coupling: the concept and clues from chromaffin and other cells

Evidence that the secreting adrenal chromaffin cell releases catecholamines directly from ATP‐rich granules

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