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  • 1
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Journal of Molecular and Cellular Cardiology 18 (1986), S. 931-941 
    ISSN: 0022-2828
    Keywords: Acidosis ; Calcium ; Diltiazem ; Energy metabolism ; Ischaemia ; Isolated rat heart ; Reperfusion ; ^3^1P NMR
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1912
    Keywords: A2 Adenosine receptor ; Human platelets ; Radioligand binding ; Adenylate cyclase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Human platelet membranes were solubilized with the zwitterionic detergent CHAPS (3-[3-(cholamidopropyl)dimethylammonio]-1-propanesulfonate) and the solubilized extract subjected to gel filtration. Binding of the adenosine receptor agonist [3H]NECA (5′-N-ethylcarboxamidoadeno-sine) was measured to the eluted fractions. Two [3H]NECA binding peaks were eluted, the first of them with the void volume. This first peak represented between 10% and 25% of the [3H]NECA binding activity eluted from the column. It bound [3H]NECA in a reversible, saturable and GTP-dependent manner with an affinity of 46 nmol/1 and a binding capacity of 510 fmol/mg protein. Various adenosine receptor ligands competed for the binding of [3H]NECA to the first peak with a pharmacological profile characteristic for the A2 adenosine receptor as determined from adenylate cyclase experiments. In contrast, most adenosine receptor ligands did not compete for [3H]NECA binding to the second, major peak. These results suggest that a solubilized A2 receptor-GS protein complex of human platelets can be separated from other [3H]NECA binding sites by gel filtration. This allows reliable radioligand binding studies of the A2 adenosine receptor of human platelets.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1912
    Keywords: Adenosine receptors ; Adenylate cyclase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary 2-Chloro-N6-cyclopentyladenosine (CCPA) was synthesized as a potential high affinity ligand for A1 adenosine receptors. Binding of [3H]PIA to A1 receptors of rat brain membranes was inhibited by CCPA with a K i-value of 0.4 nM, compared to a K i-value of 0.8 nM for the parent compound N6-cyclopentyladenosine (CPA). Binding of [3H]NECA to A2 receptors of rat striatal membranes was inhibited with a K i-value of 3900 nM, demonstrating an almost 10,000-fold A1-selectivity of CCPA. CCPA inhibited the activity of rat fat cell membrane adenylate cyclase, a model for the A1 receptor, with an IC50-value of 33 nM, and it stimulated the adenylate cyclase activity of human platelet membranes with an EC50-value of 3500 nM. The more than 100-fold A1-selectivity compares favourably with a 38-fold selectivity of CPA. Thus, CCPA is an agonist at A1 adenosine receptors with a 4-fold higher selectivity and 2-fold higher affinity than CPA, and a considerably higher selectivity than the standard A1 receptor agonist R-N6-phenylisopropyladenosine (R-PIA). CCPA represents the agonist with the highest selectivity for A1 receptors reported so far.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 3 (1969), S. 184-193 
    ISSN: 1432-0827
    Keywords: Mitochondria ; Calcification ; Calcium ; Chondrocytes ; Growth plate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des cultures d'épiphyse prélevées sur des rats et des souris ont été étudiées sous le microscope électronique. Un profile de granules mitochondriques de densité électronique a été trouvé. Les chondrocytes dans la zone proliférative avaint peu de granules, alors que ceux des zones successives ont montré une augmentation progressive de leur nombre et densité jusqu'à ce que la zone de calcification provisoire ait été atteinte. Cette zone a montré une distribution périphérique de mitochondries et une réduction du nombre et de la densité des granules mitochondriques. Du calcium isotopique 47 a été utilisé autoradiographiquement pour déterminer la location de calcium dans ces cellules. Des grains ont été trouvés sur les membranes R.E. et sur la plupart des mitochondries. La preuve d'un profile de ces granules et de leur rapport spatial avec la face de minéralisation indique une action éventuelle de mitochondries dès le début de la calcification de la matrice.
    Abstract: Zusammenfassung Epiphysekulturen von Ratten und Mäusen wurden unter dem Elektronenmikroskop untersucht. Ein Profil von mitochondrischen Körnchen mit elektronischer Dichte wurde gefunden. Chondrozyten in der Proliferationszone wiesen wenig Körnchen auf, während die der nachfolgenden Zonen allmählich an Zahl und Dichte zunahmen, bis die Zone der provisorischen Verkalkung erreicht wurde. Diese Zone zeigte eine periphere Verteilung der Mitochondrien und eine Abnahme in Zahl und Dichte der mitochondrischen Körnchen. Isotopes Kalzium 47 wurde autoradiographisch verwendet, um die Lage des Kalziums in diesen Zellen zu bestimmen. Körnchen wurden auf den E.R.-Membranen und auf einem Großteil der Mitochondrien gefunden. Der Nachweis eines Profils dieser Körnchen und ihres räumlichen Verhältnisses zur Mineralisierungsfläche weist auf einen möglich Einfluß der Mitochondrien mit Beginn der Matrixverkalkung hin.
    Notes: Abstract Rat and mice epiphyseal growth plates were studied with the electron microscope. A gradient of mitochondrial electron-dense granules was found. Chondrocytes in the proliferative zone had few granules, while those of the succeeding zones showed a gradual increase in number and density until the zone of provisional calcification was reached. This zone showed a peripheral distribution of mitochondria and a decrease in the number and density of mitochondrial granules. Isotopic47calcium was used autoradiographically to determine the location of calcium in these cells. Grains were found over the endoplasmic reticulum membranes and over most mitochondria. The demonstration of a gradient of these granules and their spatial relation to the mineralization front suggests a possible involvement of mitochondria in the onset of matrix calcification.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0827
    Keywords: Absorption ; Calcium ; Vitamin D ; Rickets ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La mise en évidence intracellulaire du calcium au niveau du revêtement muqueux de l'intestin grêle de rats normaux, rachitiques et traités à la vitamine D2 est réalisée à l'aide de l'auto radiographie au45Ca, de la micro-incinération et de la microscopie électronique. De petits granules denses aux électrons et le marquage autoradiographique sont visibles surtout dans les microvillosités et les mitochondries. Les granules sont rares dans les mitochondries des rats rachitiques, alors que les mitochondries du rat normal en sont toujours pourvues. Les mitochondries des rats, traités à la vitamine D2, contiennent de nombreux granules denses. Ces granules subsistent après micro-incinération. Un rapport inverse est noté dans le nombre des granules présents dans les mitochondries et les microvillosités. Les granules sont limités aux microvillosités chez les animaux rachitiques, et ils augmentent significativement dans les mitochondries après traitement à la vitamine D2. Cette étude indique qu'une quantité importante de calcium est absorbée à travers la membrane cellulaire et ce calcium se lie aux microvillosités. La vitamine D semble nécessaire pour mobiliser le calcium lié aux microvillosités. Ce calcium pénètre dans les mitochondries et traverse la cellule pour aller vers les vaisseaux sanguins adjacents.
    Abstract: Zusammenfassung Die intrazelluläre Lokalisation von Calcium in der Mucosa des Dünndarms von normalen, rachitischen und Vitamin D2-behandelten Ratten wurde anhand von45Ca-Autoradiographie, Microveraschung und Elektronenmikroskopie untersucht. Kleine elektronenundurchlässige Granula und autoradiographische Markierung wurden vorerst innerhalb der Microvilli und Mitochondrien festgestellt. Es fanden sich spärlich Granula in den Mitochondrien rachitischer Ratten, während sie in denjenigen von normalen Ratten regelmäßig vorlagen. Die Mitochondrien der Vitamin D2-behandelten Ratten enthielten zahlreiche elektronenundurchlässige Granula. Diese Granula waren auch nach Mikroveraschung noch vorhanden. Die Anzahl der Granula in den Mitochondrien und in den Microvilli standen in einem umgekehrten Verhältnis zueinander. Die Granula waren bei rachitischen Tieren auf die Microvillus-Region beschränkt und stiegen in den Mitochondrien nach Vitamin D2-Behandlung signifikant an. Diese Untersuchung zeigt, daß eine große Calciummenge durch die Zellmembran absorbiert und dann innerhalb des Microvillus gebunden wird. Es scheint, daß Vitamin D für die Freisetzung dieses gebundenen Calciums aus den Microvilli notwendig ist. Das freigesetzte Calcium tritt in die Mitochondrien ein oder wandert durch die Zelle zu den anliegenden Blutgefäßen.
    Notes: Abstract The intracellular localization of calcium in the mucosal lining of the small intestine of normal, rachitic, and Vitamin D2 treated rats was studied using45Ca-autoradiography, microincineration, and electron microscopy. Small, electron-dense granules and autoradiographic label were seen primarily within microvilli and mitochondria. The granules were sparse in mitochondria from rachitic rats while normal rat mitochondria demonstrated them regularly. The mitochondria of Vitamin-D2-treated rats contained numerous electron dense granules. These granules remained following microincineration. An inverse relation in the amount of granules appearing in the mitochondria and in the microvillus was found. Granules were limited to the microvillus region in the rachitic animals and increased significantly in the mitochondria following treatment with Vitamin D2. This study indicates that a large amount of calcium is absorbed across the cell membrane and is then bound within the microvillus. Vitamin D is apparently necessary for mobilizing this bound calcium from the microvilli. The mobilized calcium enters the mitochondria or passes through the cell to the adjacent blood vessels.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 5 (1970), S. 91-99 
    ISSN: 1432-0827
    Keywords: Rickets ; Cartilage ; Vitamin D ; Calcium ; Phosphate ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Abstract Des métaphyses de rats rachitiques, alimentés par paires avec des rats témoins, normaux et traités à la vitamine D2 et des phosphates, ont été étudiées au microscope électronique. La répartition des granules mitochondriales est modifiée dans le tissu rachitique: des granules ne sont visibles que dans quelques cellules voisines de la région en voie de calcification. Les rats témoins présentent une répartition, en gradient, à travers toute la métaphyse. L'adjonction à ce régime cariogène de phosphate ou vitamine D2 ou les deux à la fois, permet de rétablir la répartition normale des granules, visible chez les témoins. Il semble qu'une des modifications spécifiques, induites dans les mitochondries des chondrocytes, par un régime pauvre en phosphate et déficient en vitamine D2, est la diminution de la formation de granules mitochondriales, contenant des produits inorganiques.
    Abstract: Zusammenfassung Wachstumszonen von rachitischen Ratten werden unter dem Elektronenmikroskop verglichen mit denjenigen von 3 Kontrollgruppen (normale Diät rachitogene Diät mit Vitamin D2 und Phosphatzusätzen und eingeschränkte Menge normaler Diät). Die Verteilung der Granula in den Mitochondrien war im rachitischen Gewebe verändert; Granula wurden nur in einigen Zellen festgestellt, welche an die Zone der provisorischen Kalzifikation angrenzten. Kontrollratten zeigten einen Gradienten von Granula auf der ganzen Wachstumsplatte. Ergänzung der rachitogenen Diät durch Phosphat, Vitamin D2 oder beide bewirkte die Wiederherstellung der Granula-Dichte und-Verteilung, welche die Kontrolltiere aufwiesen. Es wird vorgeschlagen, daß die verminderte Fähigkeit, mineralhaltige Mitochondrien-Granula zu bilden, eine spezifische Veränderung ist, welche in den Mitochondrien von Chondrocyten durch eine phosphatarme, Vitamin D2-defizitäre Diät herbeigeführt wird.
    Notes: Abstract Growth plates of rachitic, pair-fed control, normal and Vitamin D2- and phosphate-treated rachitic rats were studied with the electron microscope. Mitochondrial granule distribution was modified in the rachitic tissue; granules were noted only in a few cells adjacent to the zone of provisional calcification. Control rats demonstrated a gradient of granules throughout the growth plate. Supplementation of the rachitogenic diet with either phosphate, Vitamin D2, or both was able to re-establish the granule density and distribution found in control animals. It is suggested that one specific modification induced in mitochondria of chondrocytes by a low phosphate, Vitamin D2-deficient diet is the reduced ability to form mineral-containing mitochondrial granules.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 94 (1993), S. 418-428 
    ISSN: 1432-1106
    Keywords: Red nucleus ; Motor cortex ; Reversible inactivation ; Reaching ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This study examined changes in the performance of a single-joint, elbow task produced by reversible inactivation of local regions within the proximal forelimb representation in area 4γ of motor cortex (MCx) and the red nucleus (RN) of the cat. Inactivation was carried out by microinjecting lidocaine, γ-aminobutyric acid, or muscimol into sites where microstimulation evoked contraction of elbow muscles. Reaction time, amplitude, and speed (velocity or dF/dt) of position and force responses elicited during inactivation were compared to control values obtained immediately prior to inactivation. In addition, we assessed qualitatively the effects of inactivation on reaching, placing reactions, and proprioceptive responses to imposed limb displacement. In the single-joint task, injections in MCx did not increase reaction time (simple or choice) and produced modest and inconsistent reductions in response amplitude (mean-8%) and speed (mean -19%). In contrast, injections of the same amounts of inactivating agents in the forelimb representation of RN consistently increased reaction time (34.4%), and increased the reaction time coefficient of variability (32%). There were small reductions in response amplitude (-4%) and speed (-10%) which were less than those produced by MCx inactivation. During reaching, however, these same injections in MCx and RN produced a substantial loss of accuracy. For MCx, this was due, in part, to systematic hypometria: for RN, inaccuracy resulted from increased variability in paw paths. Placing reactions and corrective responses to imposed limb displacements were also depressed by the cortical and rubral injections. Our results suggest that the forelimb representation in RN plays a role in the initiation of the single-joint, elbow tracking response examined here. The RN may mediate cerebellar regulation of response timing, a function that is likely to be important for interjoint coordination. Although neurons in the forelimb representations of MCx may contribute to force generation in single-joint movements, their contribution to multijoint control appears to be more important and is examined in the subsequent report (Martin and Ghez 1993).
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 51 (1983), S. 351-367 
    ISSN: 1432-1106
    Keywords: Cat ; Motor cortex ; Single-unit activity ; Force tracking ; Task relations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary (1) Participation of the motor cortex in initiating muscle contraction in an isometric tracking task was assessed in cats trained to make accurate force adjustments using forelimb muscles, in response to a vibrissal/visual display stimulus. Behavior in the task was characterized by short reaction times. While the task was performed, recordings of single cortical units were made in zones within area 4γ defined by the effects of microstimulation in forelimb muscles and by receptive fields on the forelimb. (2) Two types of receptive fields with different regional distributions were observed. Cells with simple receptive fields (superficial or deep) were seen throughout the area sampled, consisting of the lateral half of the anterior and posterior sigmoid gyri. Cells whose receptive fields had complex features (directional specificity, temporal lability, multiple foci, etc.) were preferentially located in the cortex rostral to the cruciate sulcus. (3) The area of motor cortex rostral to the cruciate sulcus also differed from the area caudal to the cruciate sulcus in the timing of task-related activity. Neurons that were active before response onset (lead cells), and could therefore contribute to response initiation, were preferentially located in the rostral cortex, and, in general, had complex receptive fields. (4) Lead cells were active at a constant latency from the stimulus, rather than being timed to response onset. However, the modulation of their activity was related to both the direction and magnitude of the force response. (5) These results suggest that the pericruciate motor cortex of the cat contains two functional subdivisions: a caudal one concerned with ongoing movement, perhaps under the control of specific sensory inputs from the responding limb, and a rostral one involved in initiating movement. Because behaviorally relevant stimuli can rapidly activate a specialized population of cells in the rostral cortex, this area is able to participate in responses with short reaction times.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 57 (1985), S. 427-442 
    ISSN: 1432-1106
    Keywords: Cat ; Motor cortex ; Single unit activity ; Tracking ; Input switching
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In a previous study in the cat, we have reported that motor cortex neurons discharging before the initiation of an aimed forearm response (lead cells) are better timed to movement of a display (stimulus) than to the response. The present study was done to distinguish the coding of stimulus and response features in the discharge patterns of such early activity in motor cortex. Single neurons were recorded in the arm area of motor cortex in three cats performing the same pair of responses (forearm flexion and extension) but to display movements in either of the two directions by changing display polarity. The modulation of lead cell activity was contingent on the occurrence of the learned motor response and timed to the stimulus in all conditions. The majority of lead cells (88%, n = 50) fell into one of two distinct classes. In one class of neurons, force-direction (56%, n = 32), activity was contingent on a single direction of forelimb response (flexion or extension) and was thus independent of the direction of the display stimulus. The only muscles whose patterns matched the activity of this class of response-related neurons were forelimb flexors and extensors. In these neurons, the onset of modulation was timed to one or the other of the two stimuli according to the stimulus direction which elicited the appropriate response. Thus, the display-related input to these neurons varied according to the response required. In the second class of neurons, stimulus-direction (32%, n = 18), modulation was associated with a specific stimulus direction rather than the response direction. The pattern of activity of these neurons was similar to the pattern of EMG signals of shoulder and neck muscles during the different task conditions. The contraction of proximal and axial muscles corresponded to a second response elicited by the stimulus, namely attempts at head rotation towards the moving display and was independent of the conditioned forelimb response in both time of onset and direction. To test the possibility that stimulus-direction neurons participated in the control of head rotation we trained two of the animals to also produce isometric changes in neck torque in the direction of the moving display without making the forelimb response. The activity of stimulus-direction neurons was similarly modulated during performance of the neck task. By contrast, force-direction neurons examined during the neck task were either unmodulated or discharged after the neck response. These data suggest that force-direction neurons participate in response initiation and that their activity is triggered by stimuli specific for the task. The reorganization of the inputs to motor cortex is likely to result from gating mechanisms associated with behavioral set. Such neural gates could provide for the efficient transfer of any member of an array of behaviorally relevant stimuli to restricted sectors of the somatotopically organized motor areas.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 85 (1991), S. 373-388 
    ISSN: 1432-1106
    Keywords: Red nucleus ; Motor cortex ; Single unit activity ; Tracking ; Reaction time ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In the present study we recorded the activity of single neurons in the forelimb area of red nucleus (RN) during performance of three step-tracking tasks designed to dissociate the coding of stimulus and response variables in the discharge of recorded neurons. In two of these tasks, the standard and stimulus-reversal arm tasks, elbow flexion and extension were elicited by different stimuli enabling us to distinguish activity correlated with the forelimb response from the stimulus eliciting it. The third task (neck task) allowed us to determine whether neuronal modulation was related to an unconditioned orienting response that occurred concurrently with the forelimb response. We have previously reported that these three tasks separate neurons in MCx whose modulation precedes the response (lead cells) into three distinct classes in which task-related activity either is correlated with the direction of the forelimb response, correlated with the stimulus, or not correlated with either (Martin and Ghez 1985). All lead cells, however, remained timed to the stimulus rather than to the response. The present results show that RN lead cells can be subdivided into the same three classes as those in MCx and their discharge was also contingent on the subsequent production of a behavioral response. (1) Force-direction neurons (35%; n = 16) showed changes in activity correlated with the production of forearm force in a particular direction suggesting that they could participate in selecting the appropriate forelimb response. The onset of task-related modulation of activity was better timed to the response, in contrast to force-direction neurons in MCx, which were better timed to the stimulus. (2) Stimulus-direction neurons (18%; n = 8) modulated their activity in relation to a particular stimulus evoking either flexor or extensor responses and during neck task performance. These neurons could be involved in processing stimulus information or in the production of neck torque. The task-related discharge of these lead cells was better timed to the stimulus than to either the forelimb or the neck response. (3) Nondirectional neurons (47%; n = 21) modulated their activity during all tasks examined. Their discharge did not correlate with any specific feature of the stimulus or response, and as a group, was better timed to the stimulus than to the response. Nondirectional neurons may participate in some aspect of motor preparation. To determine the relative contributions of RN and MCx lead cells to response initiation, we compared the amount of response latency variance that could be explained by variation in the latency of the unit modulation to the stimulus for the present data and the data in the earlier MCx study (Martin and Ghez 1985). Between 38% and 53% of response latency variance (for trials examined during performance of the standard arm and stimulus reversal tasks) was accounted for by the latency variations of RN force direction neurons; in contrast, 8% and 11% for MCx force-direction neurons. Variations in timing of stimulus-direction neurons in both RN and MCx account for less than 10% of response latency variance. Our findings suggest that, in the tasks examined, RN force-direction neurons play a more direct role than MCx force-direction neurons in initiating and selecting responses to stimuli. We hypothesized that this subcortical control reflects the high degree of stereotypy of the motor response examined.
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