Histoenzymatic characterization of sub-types of Type I fibres in fast muscles of rats

Summary The histochemical activities of succinic dehydrogenase (SDH), myofibrillar Adenosine triphosphatase (ATPase) and alpha glycerophosphate dehydrogenase were studied in serial sections of rat vastus lateralis (red) (RVL), gastrocnemius and diaphragm muscles. Three main fibre-types were distinguished. The Type I fibres of RVL and gastrocnemius muscles fell into two distinct groups: one category-Type IA showed very low ATPase activity. The second category of Type IB fibres displayed moderate ATPase reaction. The Type IA fibres were divisible into two sub-groups when tested for SDH reaction. Type IA₁ fibres possessed a homogenous distribution of diformazan·granules throughout the fibre: Type IA₂ fibres displayed characteristic moth-eaten pattern of diformazan localization. The diaphragm muscle did not show either Type IB or Type IA₂ varieties. The great majority of Type I fibres were sub-type IA₁ in the three fast muscles studied. It is also demonstrated here that an inherent heterogeneity exists between Type I fibres of diaphragm and leg muscles in regard to -GPD localization. This histochemical data emphasizes the fact that subdivision of Type I striated muscle fibres of mammalian animals into two sub-types is only approximate and that a further subcategorization is possible.     

Electron microscopic studies on the adenohypophysis of the White-Crowned sparrow,Zonotrichia leucophrys gambelii

Summary The pars distalis of the adenohypophysis of normal (78), thyroideotomized (6), adrenalectomized (6), and castrated (14) White-crowned Sparrows were observed with the electron microscope. Six types of glandular cells were identified and the ultrastructural characteristics of each have been described. To each has been assigned tentatively an endocrine function.STH cells are characterized by the presence of large, dense secretory granules ranging from 220–280 m, a poorly developed endoplasmic reticulum, and a fragmented Golgi apparatus; they occur only in the caudal lobe. They show no remarkable changes after adrenalectomy, castration, and thyroidectomy.Prolactin cells, whose identity is suggested by their responses to photostimulation and surgical experiments, are characterized by large, polymorphic, dense secretory granules; they have been found mainly in the cephalic lobe.ACTH cells, whose function is confirmed by their cytological responses to adrenalectomy, have a peculiar type of secretory granule (220 m) with high and low phases of electron density. They occur exclusively in the cephalic lobe and are transformed, after adrenalectomy to large, vacuolated adrenalectomy cells.TSH cells are so designated by their response to thyroidectomy. After thyroidectomy, they lose their specific fine secretory granules and are transformed into large, vacuolated thyroidectomy cells. We have found TSH cells and thyroidectomy cells only in the cephalic lobe.Two types considered to be gonadotropic cells from their responses to gonadectomy, occur in both the cephalic and caudal lobes. One of them contains spherical, dense secretory granules (180–220 m), prominent rough endoplasmic reticulum and a well developed Golgi apparatus; the other type contains dense secretory granules of variable size (150–350 m), a less extensively developed Golgi apparatus, and sac-like endoplasmic reticulum. Both types of gonadotropic cells show extreme enlargement and vacuolization after castration. However, they retain differences in appearance in the structure of cytoplasmic organelles and vacuolization.Dedicated to Professor Dr. H. Grau in honor of his 70th birthday.The investigation reported herein was supported by a research grant (HE 07240 NEUA) from the National Institutes of Health to Professor Vitums, by a research grant (5R01 NB 06187) from the National Institutes of Health to Professor Farner, and by a scientific research grant (No. 91049) from the Ministry of Education of Japan to Professor Mikami. The authors wish to thank Professor James R. King for his assistance in obtaining and maintaining the birds, and for his helpful advice concerning the experiments.     

Current chemical concepts of acids and bases and their application to anionic (“acid”) and cationic (“basic”) dyes

Summary In biomedical studies, dyes are divided into acid and basic dyes. This classification cannot be reconciled with current chemical definitions of acids and bases. Brönsted-Lowry acids are compounds that can donate protons; bases are proton acceptors. The definition of acids and bases is independent of the electric charge, i.e. acids and bases can be neutral, anionic or cationic. Reactions between acids and bases result in formation of new acid-base pairs. Lewis acids and bases do not depend on a particular element, but are characterized by their electronic configurations. Lewis bases are electron donors; Lewis acids are electron acceptors. This classification is also unrelated to the electric charge. Lewis acids and bases interact by formation of coordinate covalent bonds.In histochemistry and histology, dyes containing SO ₃ ^– , –COO^– and/or –O^– groups are classified as acid dyes. However, such compounds are electron pair donors and hence Brönsted-Lowry and Lewis anionic bases. Dyes carrying a positive charge are termed basic dyes. Chemically, many cationic dyes are Lewis acids because they can add a base, e.g. OH^–, acetate, halides. The hypothesis that transformation of –NH₂ into ammonium groups imparts basic properties to dyes is untenable; ammonium groups are proton donors and hence acids. Furthermore, conversion of an amino into an ammonium group blocks a lone electron pair and the color of the dye changes drastically, e.g. from violet to green and yellow. It appears therefore highly unlikely that ammonium groups are responsible for binding of cationic (basic) dyes. In histochemistry, it is usually not of critical importance whether anionic or cationic dyes are chemically acids or bases. It is therefore suggested to substitute the terms anionic for acid and cationic for basic dyes; this nomenclature will always be chemically correct.     

Photoreversion und Photoprotektion bei UV-induzierten Mutationen des nichtphotoreaktivierbaren Bacteriums E.coli phr?

Summary Stationary phase cells of strain phr^–/MC 2 ofE. coli are not photoreactivable but the frequency of UV-induced mutations to low Streptomycine-resistance (S 3, 3/ml) is decreased strongly by illumination with light of fluorescence tubes (310 to 500 nm) after UV-irradiation. Also dark-reversion (DRM) of these mutations due to keeping UV-irradiated cells in saline is observed. Illumination before UV-irradiation decreases the frequency of the mutations (photoprotection against mutation=PPM) to the same extent as the combined action of photoreversion (PRM) and DRM. The lag-phase of cell division is prolonged strongly by illumination from 80 min without light to 150 min by the light-dose of highest activity. The additional lag is nearly the same if the illumination is done before, after or without UV-irradiation; this lag is about additive to the small lag caused by UV. Pre-illumination of the stationary-phase cells does not cause photoprotection against killing (PP), it even decreases the survival after high UV-doses. The observations support the hypothesis that PRM in this strain may be indirect, i.e. caused by the light-induced additional division lag which enhances the dark repair of UV-premutations. Also spontaneous premutations which are apparently present in the stationary-phase cells seem to be influenced by the light in this way.     

Die Hypokotylfarbe als Markierungsfaktor von Genomstufen der Zuckerrübe

Zusammenfassung In zunehmendem Maße werden anisoploideBeta-Rübensorten angebaut, deren zytologische Kontrolle zwecks Feststellung der Genomstufenprozentanteile recht arbeitszeitaufwendig ist. Übereinstimmend mit polnischen Autoren wurde festgestellt, daß die Hypokotylfarbe ein geeigneter Markierungsfaktor für die einzelnen Genomstufen darstellt. Kreuzt man tetraploide Pflanzen, die ein grünes Hypokotyl besitzen, mit diploiden Pflanzen, die ein rosa Hypokotyl aufweisen, so erhält man von dem tetraploiden Partner tetraploide grüne und triploide hellbraune, von dem diploiden Partner diploide rosa und triploide hellbraune Nachkommenschaften. Die in bezug auf die Hypokotylfarbe heterozygoten Pflanzen kann man demnach von den homozygot grünen und homozygot rosa Individuen unterscheiden. Die Kreuzung diploid grünxtetraploid rosa ist für diese Zwecke nicht brauchbar, da sich die triploiden Heterozygoten mit einem grünen und zwei rosa Allelen in der Hypokotylfarbe nicht deutlich von den homozygoten rosa Pflanzen abheben. Auf die Bedeutung dieser Markierungsmöglichkeit für bestimmte Forschungsprobleme, die Züchtung und die Saatgutkontrolle wird hingewiesen.     

The cytology of phytophthora infestans

Summary Phytophthora infestans has three kinds of somatic nuclei: an oval shaped nucleus (approx. 3.1×2.7 ) which stains diffusely except for a crescent shaped Feulgen positive cap which stains intensely; a granular nucleus whose contents are organized into a fairly constant number of stained bodies, and, a deeply staining condensed nucleus. The capped nucleus is thought to be metabolic or resting and the granular nucleus is thought to be dividing as it is most commonly found in hyphal tips. Attenuated forms of all three kinds of nuclei are found.Nuclear division is mitotic and intranuclear. Eight—ten chromosomes are seen at metaphase.Sporangia have a mean of 6.3 nuclei which is constant for age and strain of culture. Sporangia become multinucleate as a result of nuclear migration and not by division in the developing sporangium. Zoospores are usually uninucleate.The nuclear cap is persistent throughout nuclear division when it also divides. It is associated with flagella production and nuclear migration and has some of the properties of a blepharoplast.     

Computational constraints that may have favoured the lamination of sensory cortex

At the transition from early reptilian ancestors to primordial mammals, the areas of sensory cortex that process topographic modalities acquire the laminar structure of isocortex. A prominent step in lamination is granulation, whereby the formerly unique principal layer of pyramidal cells is split by the insertion of a new layer of excitatory, but intrinsic, granule cells, layer IV. I consider the hypothesis that granulation, and the differentiation between supra- and infra-granular pyramidal layers, may be advantageous to support fine topography in their sensory maps. Fine topography implies a generic distinction between where information, explicitly mapped on the cortical sheet, and what information, represented in a distributed fashion as a distinct firing pattern across neurons. These patterns can be stored on recurrent collaterals in the cortex, and such memory can help substantially in the analysis of current sensory input. The simulation of a simplified network model demonstrates that a non-laminated patch of cortex must compromise between transmitting where information or retrieving what information. The simulation of a modified model including differentiation of a granular layer shows a modest but significant quantitative advantage, expressed as a less severe trade-off between what and where. The further connectivity differentiation between infra-granular and supra-granular pyramidal layers is shown to match the mix of what and where information optimal for their respective target structures.     

We Do Not Dream of the 3 R's: Implications for the Nature of Dreaming Mentation

This report examines the extent to which dream recall involves the 3 R's (reading, writing, and arithmetic). Two separate studies were done. In the first study, two scorers rated, on a blind basis, a total of 456 written dream reports, available from five previous studies. There was perfect agreement between the two scorers. They agreed that there were no instances of reading, no instances of writing, and one instance of probable calculating in the 456 dreams. The second study was a questionnaire survey. Complete responses were obtained from 240 frequent dreamers (who reported remembering a mean of seven dreams per week). The study examined in two ways the frequency of the 3 R's in their recalled dreams. First, in answer to direct questions as to how frequently they dreamt about each activity, roughly 90% of the respondents reported that they never or hardly ever dreamt about each of four activities: reading, writing, typing, and calculating. In answers to other questions, this group reported spending a mean of six hours per day engaged in these activities. Second, responses as to the relative prominence of six activities (walking, writing, talking with friends, reading, sexual activity, typing) in dreaming versus waking produced two clear groupings of activities. Walking, talking with friends, and sexual activity were each rated almost as prominent in dreaming as in waking whereas the second group consisting of writing, reading, and typing were rated as far more prominent in waking than in dreaming. The two activity groups differed at p < .0001. Thus, the 3 R's appear to occur very infrequently in dreams. These findings are placed in a theoretical frame which suggests that dreaming (compared to waking) deals very little with serial activities characterized by input—rapid-processing—output in which the neural nets function in a feed-forward mode. Rather, dreaming may be characterized by relatively broad or loose connection making in which the nets function more in an autoassociative mode.     

Methodological problems in evolutionary biology VIII. Biology and culture

Biology cannot accommodate all aspects of culture. Aspects of culture that a biological approach can take into account can be covered by the biological categories of phenotype and environment. There is no need to treat culture as a separate category. Attempts to elaborate biological explanations of cultural variation will meet with success only if biologists expand theories of development, and integrate them in evolutionary biology. The alternative — elaborating the idea of so-called cultural inheritance — makes little sense from a biological point of view.     

„Kolbenbildungen“ der Muskelfasern des menschlichen Kehlkopfs

Zusammenfassung Fasern der menschlichen Kehlkopfmuskulatur weisen seitliche Aussackungen auf, die unter dem Sarkolemm in lockerer Anordnung Kerne, Lysosomen, Lipofuscingranula und Mitochondrien mit oft konzentrisch angeordneten Cristae enthalten. Sie sind mit den lichtmikroskopisch bekannten End- bzw. Seitenkolben identisch. Diese Aussackungen werden von den gefäßnahen subsarkolemmalen Mitochondrienanhäufungen eines bestimmten, auch in der übrigen Skelettmuskulatur weitverbreiteten Muskelfasertyps abgeleitet. Es wurden keine Verbindungen mit Nervenfasern gefunden. Im Gegensatz zu Goerttler können wir diese Bildungen deshalb nicht als Mechanorezeptoren deuten. Die Endkolben werden als sekantial getroffene Seitenkolben aufgefaßt. Da die Aussackungen der Fasern auch bei gesunden jungen Männern in der Kehlkopfmuskulatur regelmäßig auftreten, handelt es sich hier nicht um eine krankhafte Bildung, wenn auch die Faser im Bereich der Aussackungen einen degenerativen Prozeß durchzumachen scheint.

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Summary Fibers of human larynx muscles show lateral dilatations of the sarcolemma containing loosely disposed mitochondria, lysosomes and eventually one or more nuclei. The cristae of the mitochondria often are arranged concentrically. Corresponding fiber dilatations have already been described by light microscopists as side- resp. end-bulbs.Our electron-microscopical observations lead to the conclusion that the dilatations are derived from accumulations of mitochondria beneath the sarcolemma which in similar form — but smaller — are characteristic of one type of the different muscle fibers, frequently to be found in other muscles of the body. Both subsarcolemma accumulations of mitochondria are close to the muscle capillaries.The fiber dilatations have no connections with nerve axons, and therefore cannot be interpreted as mechanoreceptors (Goerttler, 1960). The end-bulbs are supposed to be secantially (or obliquely) cut side-bulbs. We do not consider them pathologic since they are regularly found in fibers of healthy young men. On the other hand, they may represent a circumscribed degenerative alteration the origin of which remains to be explored.

     

Variant forms ofα-2-l-fucosyltransferase in human submaxillary glands from blood group ABH “secretor” and “non-secretor” individuals

The properties of the -2-l-fucosyltransferases in submaxillary gland preparations from blood group ABH secrefors and non-secretors were compared. The level of activity in the non-secretor gland homogenates amounted to about 5% only of that found in the secretor gland preparations. The enzymes from the two sources differed in solubility properties, charge and affinities for donor and acceptor substrates. The enzyme from secretor glands showed a preference for acceptors with Type 1 [d-galactosyl(1–3)-N-acetyl-d-glucosamine] structures whereas the enzyme from non-secretor glands had a preference for Type 2 [d-galactosyl(1–4)-N-acetyl-d-glucosamine] structures.These results demonstrate that expression of the secretor gene (Se) is associated with a molecular form of the -2-l-fucosyltransferase that is different from the species present in the same tissue when theSe gene is not expressed.     

Circadian changes in synaptic ribbons and spherules in pinealocytes of the Syrian hamster (Mesocricetus auratus)

Summary In the present study, synaptic ribbons were studied morphologically and quantitatively in hamster pineal gland. The number of ribbons and spherules of hamster pinealocytes was counted over a 24-h period. The 24-h variations in the quantity of synaptic ribbons were found to parallel fluctuations in pineal melatonin concentrations. No significant circadian changes were observed for synaptic spherules, indicating different roles for these two structures.     

Some observations on the fine structure of the myotendinous junction in myotomal muscles of the tadpole tail

Summary Myotendinous junctions in the myotomal tail muscles of the tadpole of Rana rugosa were examined by electron microscopy. At the site of the myotendinous junction, the sarcolemma is covered on its sarcoplasmic aspect by the connecting filament layer and the attachment layer, and on the extracellular aspect by the intermediary layer and the external lamina, with associated collagen fibrils. The intermediary layer consists of filamentous structures which closely resemble microfibrils (Hanak and Böck, 1971), spine-like or thread-like profiles (Korneliussen, 1973) and intermediary layer (Nakao, 1975a, b) in the myotendinous junctions of other vertebrate skeletal muscles.Particularly interesting is the fact that all the coverings and linings of the sarcolemma, including the external lamina, are completely absent in the terminal segment of the finger-like sarcolemmal invagination characteristic of the myotendinous junction. Furthermore, special types of coupling between a sac of sarcoplasmic reticulum and a part of the sarcolemmal invagination are frequently observed. These couplings always occur along the region of the sarcolemma where the external lamina is absent. The couplings show features similar to those of the triad, such as SR feet , scalloped SR membranes and granular content of the SR sac, suggesting that they are analogous and functionally similar to the triad and other equivalent structures.     

Die Richtungen des Nahrungsflusses im Darmtrakt der Kleinzikade Euscelidius variegatus KBM. (Jassidae)

Zusammenfassung Der Verlauf des Nahrungsflusses im Darmtrakt der Kleinzikade Euscelidius variegatus wird nach Verfütterung von farbstoffhaltiger Nährlösung ermittelt. Es wird der Beweis erbracht, daß die aufgenommene Nahrungsmenge in der Filterkammer geteilt wird und die beiden Anteile den Darmtrakt auf zwei verschiedenen Wegen in Richtung Rektalblase passieren. Ein Anteil der aufgenommenen Nährlösung wird über einen Kurzschlußweg in der Filterkammer sowohl über den Filterkammerdarm als auch über die Kryptonephridien direkt in den Enddarm gepumpt, während die in der Magentasche der Filterkammer verbleibenden Nahrungsanteile über einen langen Verdauungsweg zum After gelangen. Hierbei wird der Magentascheninhalt in den Magen gedrückt. Von dort aus passiert er den Mitteldarm und erreicht über den Enddarm den After. Der Kurzschlußweg und der Verdauungsweg können gleichzeitig benutzt werden. Der Kurzschlußweg wird von der Nahrung jedoch in viel kürzerer Zeit durchströmt als der längere Verdauungsweg.

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The directions of the flow of food in the alimentary trad of the leafhopper Euscelidius variegatus KBM. (Jassidae)

Summary The leafhopper Euscelidius variegatus is fed with synthetic food, coloured with 1% Azorubin-S. Its flow in the alimentary tract has been studied. It has been found that the sucked-in food is divided into two parts in the filter chamber, each taking different way in the alimentary tract for its flow. One part of the food is pumped into the hindgut via the short circuit way going through the filter chamber once over the Filterkammerdarm and also over the kryptonephries. That part of the food, which remains in the pocket of the filter chamber takes the long digestion way to the anus over stomach, midgut and hindgut. Both the ways could be used at the same time. But the food takes much shorter time for its passage through the short circuit way as compared to the time needed for the long digestion way.

     

Intra- and extraganglionic peripheral cholinergic neurons in the urogenital organs of the cat

Summary The distribution of cholinergic neurons in the urinary tract and male genital organs of the cat was studied by a histochemical method for acetylcholinesterase. In addition to cell clusters in autonomic ganglia (intraganglionic cells), isolated extraganglionic cholinergic cells were found within the innervated tissues, usually in association with nerve trunks and blood vessels. Smaller neural cells with multiple axonal processes, identical to Cajal's interstitial cells, were found in the meshes of the terminal nerve plexus in smooth muscle, lamina propria and vascular wall.It is concluded that peripheral cholinergic neurons, like their adrenergic analogues, are arranged as a short intraganglionic, a shorter extraganglionic, and a terminal system of neurons.Supported in part by grants 10465 and 11285 from the USPHS and the Henry C. Buswell Urology Research Fund.     

Ultrastructure of four types of striated muscle fibers in the atlantic hagfish (Myxine glutinosa,L.)

Summary Four types of striated muscle fibers with distinctive ultrastructure were defined in the Atlantic hagfish (Myxine glutinosa, L.): white, intermediate, and red fibers of m. parietalis, and red fibers of m. craniovelaris.White fibers are thick, contain very few mitochondria and fat vacuoles, and possess distinct and separate myofibrils with thin Z-disks and distinct M-lines. Intermediate fibers are thinner, possess largely similar myofibrils that often are even better separated due to a higher content of fat vacuoles and especially mitochondria and glycogen granules. Red fibers of m. parietalis contain large amounts of mitochondria, fat vacuoles, and glycogen granules. Their myofibrils possess M-lines, and although branching more, the myofibrils of red fibers conform with a Fibrillenstruktur pattern like those of white and intermediate fibers. Red fibers of m. craniovelaris are very thin, possess many smaller fat vacuoles, and large amounts of mitochondria and glycogen granules. The myofibrils are significantly thinner than in m. parietalis fibers, run as quite independent well separated units, possess thicker Z-disks, and lack M-lines. Large amounts of myosatellite cells are associated with these red fibers.Triads are located near A/I-junctions in all four fiber types and occur irregularly, the density of triads being different in the various fiber types.We are indebted to Dr. Finn Walvig, Biological Station, University of Oslo, Drøbak, for supply of hagfishes, and we also wish to thank Dr. Jan K. S. Jansen, Institute of Physiology, University of Oslo, for valuable suggestions during this study.     

Physicochemical and morphological characterization of two small polluted streams in southwest Germany

As a supporting component of the VALIMARproject, physicochemical investigations wereconducted monthly from 1995 to 1999 at theKrähenbach/Aich stream system (two samplingsites) and at the Körsch stream (sixsampling sites). Several physicochemicalparameters were analysed continuously bydataloggers during the entire sampling period.Moreover, a selection of the most importantmorphological parameters of the stream wererecorded in 1998.The results of these physicochemicalinvestigations demonstrated that theKrähenbach/Aich system could becharacterized as a natural rhithral submontanecarbonate stream system that is welloxygenated. Low temporal variations in pH,conductivity and chloride occurred at allsampling sites in the Krähenbach whichcoincided with the geological conditions atthis test stream. Because of relatively lowconcentrations of organic and inorganiccompounds during the entire study period thesites AB and AC in the Krähenbach/Aichsystem were classified as slightly polluted.The morphological assessment of the streamindicated a moderately affected status atboth of these streams.Physicochemical and morphologicalinvestigations along the Körsch streamsystem (KA to KE including the tributary streamSulzbach) indicated a highly affected rhithralsubmontane carbonate system characterized byhigh water velocity, flow rate and dissolvedoxygen. The Körsch, however, differssignificantly with respect to watertemperature, conductivity, pH, BOD₅,nutrients, and chloride concentrations from theKrähenbach. As a result of differentanthropogenic inputs (e.g. several sewageplants, agriculture activities and waste waterdumping) the Körsch sites KA to KD wereclassified as critically to heavilypolluted. The morphological assessmentindicated a clearly affected to damagedstatus. The upstream site KE could becharacterized as slightly polluted and itsmorphological structure as moderatelyaffected.     

Ultrastructure of cell wall and plugs of tobacco pollen tubes after chemical extraction of polysaccharides

Tobacco pollen tubes grown in vitro and from pollinated tobacco styles were treated by chemical solvents to remove one or more of the following polysaccharides from the tube walls: pectin (ethylenediamine tetraacetic acid); hemicellulose (alkali); callose (alkali; potassium hypochlorite); cellulose (cuprammonium); and all polysaccharides with exception of cellulose (H₂O₂/glacial acetic acid). Both the inner tube wall, which we had regarded as the secondary wall, and the plugs contained, in addition to callose, microfibrils of cellulose and non-cellulosic microfibrils that had pectin-like properties. When using the expressions callosic or callose layer and callose plugs in reference to pollen tubes, one should realize that they do not imply the exclusive presence of callose in the inner tube wall layer and its localized thickenings.Extended version of a contribution (poster) presented at the International Symposium Advances in Plant Cytoembryology in Lublin, Poland, in June 1980 Dedicated to Professor J. Straub (Köln-Vogelsang) on his 70th birthday in 1981     

Regulation of adenylate cyclase by adenosine

Summary Adenosine may well be as important in the regulation of adenylate cyclase as hormones. Sattin and Rall first demonstrated in 1970 that adenosine was a potent stimulator of adenylate cyclase in the brain. However, adenosine is an equally potent inhibitor of adenylate cyclase in other cells such as adipocytes. The concentration of adenosine required for this regulation of adenylate cyclase is in the nanomolar range (10 to 100 nm). Both the inhibitory and stimulatory effects of low concentrations of adenosine on adenylate cyclase are antagonized by methylxanthines. This antagonism of adenosine action may account for all or part of the effects of methyl xanthines on cyclic AMP levels in many tissues. Adenosine appears to be a particularly important endogenous regulator of adenylate cyclase in brain, smooth muscle and fat cells. Under conditions in which intracellular AMP rises, adenosine formation and release is accelerated. In addition to its direct effects on adenylate cyclase, adenosine (at higher concentrations approaching millimolar) exerts multiple effects on cellular metabolism as a result of its intracellular metabolism and especially conversion to nucleotides.The effects of nanomolar concentrations of adenosine on adenylate cyclase are mediated through an adenosine site possessing strict structural specificity for the ribose moiety of the molecule (the R adenosine site) which is presumably located on the external surface of the plasma membrane. In brain, lung, platelets, bone, lymphocytes, skin, adrenals, Leydig tumors, and coronary arteries adenosine stimulates adenylate cyclase via this site. However, in rat adipocytes, brain astroblasts and ventricular myocardium adenosine inhibits adenylate cyclase through the R or adenosine site. Although the R site requires an intact ribose moiety, adenosine analogs modified in the purine ring such as N⁶-phenylisopropyladenosine appear to be potent agonists for this site. All effects of adenosine mediated via the R site are competitively antagonized by methyl xanthines.The effects of micromolar concentrations of adenosine appear to be mediated via a site with strict structural specificity with respect to the purine moiety of the molecule (the P or adenine adenosine site). This P site is postulated to be located on the intracellular face of the plasma membrane and mediates the effects of adenosine due to conversion of adenosine to 5-AMP or perhaps other nucleotides. The effects of high concentrations of adenosine are always inhibitory to adenylate cyclase activity, are readily demonstrated in broken cell preparations, and are unaffected by methylxanthines. An intact purine ring is required for these adenosine effects but modifications of the ribose moiety of the molecule generally increases the potency of the analog. A prime example is 2,5-dideoxyadenosine, which is the most potent known R-site specific adenosine analog.We propose a unitary model which explains both the stimulatory and inhibitory effects of low concentrations of adenosine on adenylate cyclase. In brief, adenylate cyclase is postulated to exist in three interconvertible activity states: (i) an inactive state (E₀); (ii) a GTP-liganded state with high activity (E_GTP); and (iii) a GDP-liganded state (E_GDP) which is inactive in cells where adenosine stimulates adenylate cyclase, but active in cells where adenosine inhibits adenylate cyclase. We postulate that the enzyme cycles through these states in the following manner: the E₀ state binds GTP and forms the E_GTP state; hydrolysis of bound GTP converts the E_GTP to the E_GDP state; and release of bound GDP converts E_GDP to the E₀ state. The E₀ state is the only form of the enzyme which can be stimulated by either hormones or GTP and its formation from the E_GDP state is rate-limiting in this cycle. The conversion of E_GDP to E₀ regulates the ability of hormones and GTP to activate adenylate cyclase and is postulated to be adenosine sensitive.In cells where both E_GDP and E₀ states are inactive, adenosine stimulates adenylate cyclase activity. In cells where E₀ is inactive, but E_GDP is active, adenosine inhibits adenylate cyclase activity. In addition we suggest that in cells where adenosine inhibits adenylate cyclase activity (cells postulated to have an E_GDP state which is active) high concentrations of GTP favor accumulation of the enzyme in E_GDP and thus are inhibitory to activity. Prostaglandins may also regulate adenylate cyclase in a manner similar to that described above for adenosine.We conclude that adenosine is an important regulator of adenylate cyclase whose role has only been appreciated recently. Further studies are warranted on both its binding to cells and mechanisms by which it regulates adenylate cyclase.This work was supported by United States Public Health Service Research Grant AM-10149 from the National Institute of Arthritis, Metabolism and Digestive Diseases.