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C Cognard

Publications and source records attributed to C Cognard.

At least 73 records · Page 4Linked to original sources
PubMed1996

Hypoosmotic shocks induce elevation of resting calcium level in Duchenne muscular dystrophy myotubes contracting in vitro.

In Duchenne muscular dystrophy (DMD) muscle cells which lack dystrophin, contraction seems to be a dominant factor contributing to the abnormal elevated intracellular calcium level. Human normal and DMD contracting myotubes cocultured with nervous cells were exposed to a hypotonic medium to mimic contraction-induced mechanical stress on the membrane, and the cytoplasmic calcium activity was simultaneously monitored (Indo-1). Hypotonic shocks induced a reversible [Ca2+]i increase in 81% of the DMD cells vs. 54% of control. In addition, responses were qualitatively different: most of DMD myotubes displayed a fast increase of Ca2+ flowing from the edge of the myotube while the response in normal cells was slow and diffuse. The fact that these responses were not affected by ryanodine, was in favour of an external source of Ca2+ involved in the hypoosmotic shocks. The localized increase of Ca2+ in DMD myotubes, inhibited by Gd3+, could result from sites of high mechanosensitive channel activity or density which could constitute a pathway for Ca2+ entry provided these cells contract.

Calcium↗
PubMed1996

Myoblast fusion requires cytosolic calcium elevation but not activation of voltage-dependent calcium channels.

Many studies of in vitro skeletal myogenesis have demonstrated that fusion of myoblasts into multinucleated myotubes is regulated by calcium-dependent processes. Calcium ions appear to be necessary at the outer face of the membrane, and an additional internal calcium increase seems required to promote fusion of aligned myoblasts. It has been proposed that a calcium influx could take place prior to fusion and that this may be mediated by voltage-dependent calcium channels. Previously, we showed that two types of voltage-dependent calcium currents were expressed in multinucleated myotubes but not in rat myoblasts growing in primary culture before the withdrawal of the growth medium. We also showed that the previous formation of multinucleated synticia was not a prerequisite of developmental appearance of calcium currents, suggesting that the two events were time-correlated but not sequentially dependent. These features led us to investigate changes in internal calcium activity and the possible appearance of voltage-dependent calcium influx pathways just after the promotion of fusion by the change of culture medium. The results confirm that a rise in cytosolic calcium activity occurs slightly before fusion in confluent myoblasts and remained in newly formed myotubes. Reducing this elevation by internal calcium buffering lowered myoblast fusion and, reciprocally, blocking cell fusion prevented calcium increase. Treatment with the organic calcium channel blockers nifedipine (5 microM) and PN 200-110 (1 microM) did not alter cytosolic calcium changes nor cell fusion, and voltage-dependent calcium currents were never observed by the perforated patch-clamp technique in aligned fusion-competent myoblasts. Other voltage-operated mechanisms of calcium rise were not detected since depolarization with hyperpotassium solutions failed to elicit increases in intracellular calcium. On the contrary, acetylcholine was able to promote extracellular calcium-dependent calcium transients. Our results confirm the requirement of an increase in resting calcium during fusion, but do not support the hypothesis of an influx through voltage-dependent channels or other voltage-operated pathways. The elevation of internal calcium activity may result from other mechanisms, such as a cholinergic action for example.

Animals↗
PubMed1996

Intracranial dural arteriovenous fistulas with spinal venous drainage: relation between clinical presentation and angiographic findings.

PURPOSE: To investigate why some patients with an intracranial dural arteriovenous fistula (DAVF) with spinal venous drainage have myelopathy and others do not. METHODS: We reviewed the clinical and radiologic data for 12 patients who had a DAVF with spinal venous drainage diagnosed at our institutions from 1982 to 1995. RESULTS: Six patients had progressive spinal cord indications of disease (patients with myelopathy) and six others (patients without myelopathy) had cerebral indications (five had intracranial hemorrhage and one had a seizure). Cerebral angiography showed a posterior fossa DAVF with spinal venous drainage in all cases. The clinical presentation of DAVFs with spinal venous drainage was compared with the extent of the drainage. In patients without myelopathy, the spinal venous drainage exited the intradural canal via the cervical medullary-radicular veins and was therefore limited to the cervical perimedullary veins. In patients with myelopathy, no medullary-radicular vein was seen, and the venous drainage descended along the perimedullary veins of the entire spinal cord toward the conus medullaris. CONCLUSION: We found an exact relation between clinical presentation and venous drainage of DAVFs with spinal venous drainage. Patients had no myelopathy when the venous drainage was limited to the cervical cord; myelopathy was present when the venous drainage descended toward the conus medullaris.

Adult↗
PubMed1995

Myoblast fusion is not a prerequisite for the appearance of calcium current, calcium release, and contraction in rat skeletal muscle cells developing in culture.

During in vitro development of rat skeletal muscle cells, contraction and calcium currents progressively appear after fusion of myoblasts. To investigate whether muscle-specific functions are expressed in the absence of myoblast fusion, rat neonatal muscle cells were cultured in a differentiation medium under conditions that are well known to inhibit fusion: prolonged culture in a low-calcium medium or treatment with cytochalasin B. We have demonstrated that the fusion-arrested cells expressed differentiative properties in L-type calcium current, transient release of calcium ions from internal stores in response to caffeine and depolarizing agents, and contraction elicited by depolarization. Properties and potential-dependence of L-type calcium currents were similar to that in control fused cells, but T-type calcium currents were not observed, while both types coexist in myotubes. Properties of calcium transients and voltage dependence of contraction suggested that the excitation-contraction mechanisms were well established. However, comparing to well-developed myotubes at the same time of culture, the characteristics of calcium transients and contraction of fusion-arrested cells were closer to those of younger myotubes, which can be interpreted in terms of a delay in maturation of excitation-contraction coupling and contractile machinery. All these observations demonstrate that myoblast fusion is not necessary for triggering the establishment of calcium transport and release and contractile functions of rat muscle cells developing in culture. The appearance of muscle-specific functions is consistent with previous results demonstrating that the fusion-arrested cells express muscle-specific proteins and structures.

Acetylcholine↗
PubMed1995

Properties of calcium currents and contraction in cultured rat diaphragm muscle.

The characterization of calcium currents and contraction simultaneously measured in cultured rat diaphragm muscle cells was carried out in the present study. Whole-cell patch-clamp experiments were designed to further elucidate the mechanism of excitation-contraction (E-C) coupling in diaphragm which, though generally considered a skeletal-type muscle, has been reported to exhibit properties indicative of a cardiac-like E-C coupling mechanism. Normalized current/voltage (I/V) curves for two concentrations of external calcium (2.5 and 5 mM) were obtained from diaphragm myoballs. Both curves showed peaks corresponding to the activation of a T-type calcium current and a dihydropyridine-sensitive L-type calcium current. The normalized curve for the voltage dependence of the activation of contraction in diaphragm myoballs followed a typical Boltzmann-type relationship to the peak of contraction. Thereafter, the curve declined in a manner that was more pronounced in diaphragm compared to that measured in additional experiments using cultured rat limb muscle myoballs. This effect could be interpreted in terms of a more pronounced participation of the L-type current in E-C coupling in cultured diaphragm muscle. An increased likelihood of cultured diaphragm muscle to undergo depletion of sarcoplasmic reticular calcium stores during repetitive stimulation, or a heightened propensity for the voltage sensor for E-C coupling in diaphragm to enter the inactive state could also explain this effect. Maximal contractile activity was only slightly affected when the L-type current was blocked by externally applied cadmium (2 mM) or cobalt (3 mM), suggesting that a pronounced calcium-current-dependent component of contraction is unlikely in cultured diaphragm muscle. These results show that T- and L-type calcium channels are expressed in cultured rat diaphragm muscle cells and that, in contrast to cardiac muscle, the entry of calcium ions via L-type voltage-dependent calcium channels is not a prerequisite for contraction. Differences in the voltage sensitivity of contraction, observed at depolarized membrane potentials in cultured rat diaphragm and limb muscle cells, suggest that the voltage sensor for E-C coupling in diaphragm might more readily enter an inactivated configuration - possibly by a mechanism which is dependent on the concentration of external calcium.

Animals↗
PubMed1995

Abnormal calcium homeostasis in Duchenne muscular dystrophy myotubes contracting in vitro.

Resting intracellular calcium activity was recorded in three kinds of human muscle cells in culture: normal (control) and dystrophic (DMD and FSH), by means of a ratiometric fluorescence method using the calcium probe Indo-1 under laser illumination. DMD cells are characterized by a lack of dystrophin whereas FSH cells express normal dystrophin. The aim of this study was to determine whether, in dystrophin-deficient muscle cells (DMD), contraction destabilized internal calcium homeostasis. Muscle cells were cocultured with rat spinal cord explants to improve the maturation of human myotubes up to the stage where contraction appears. The resting intracellular calcium level was significantly higher in contracting DMD cells (107 +/- 8 nM; n = 44) compared to control cells (66 +/- 6 nM; n = 43) or in FSH cells (56 +/- 6 nM; n = 35). DMD myotubes cocultured in the presence of TTX which inhibited contractile activity, did not develop an increase in free cytosolic Ca2+ concentration. The amplitudes of calcium transients elicited by exposure to acetylcholine (ACh) or high K+ medium (100K) were significantly higher in contracting DMD myotubes than in control ones. The extra-responses were not observed in DMD myotubes cocultured with TTX. This study strongly suggest that: (i) contraction is a dominant factor contributing to Ca2+ abnormalities in DMD cells; and (ii) contracting dystrophin-deficient cells have defective calcium handling mechanisms during electrical events which involve sarcolemma.

Acetylcholine↗
PubMed1995

Cultured rat skeletal muscle cells treated with cytochalasin exhibit normal dystrophin expression and intracellular free calcium control.

Many studies performed to elucidate the molecular and cellular processes involved in muscular dystrophies have led to the working hypothesis of a key role for the cytoskeleton elements linking the extracellular matrix to myofibrils. It was recently suggested that cytochalasin B treatment of mouse soleus muscle promoted cell damage mediated by a cytosolic increase in free calcium concentration. Since intracellular calcium overload may be a primary event resulting from the alteration of cytoskeletal structure, this study was intended to evaluate whether or not the integrity of the F-actin microfilament network is necessary for calcium homeostasis. The developmental establishment of the normal cytoarchitecture was altered by treatment of myoblasts with the actin-disrupting agents cytochalasin B and D, and the effects were compared with those in myoblasts treated with colchicine. These drugs modified the morphogenesis in that they prevented the formation of elongated myotubes by myoblast fusion, but did not prevent the maturation of contractile myogenic cells. The subcellular organisation of actin filaments visualised by confocal fluorescence microscopy was modified by colchicine and cytochalasins, but appearance of contractile apparatus and mechanical activity were not precluded. Sarcolemmal addressing of dystrophin, the subsarcolemmal protein lacking in Duchenne muscular dystrophy, was not prevented by cytochalasin. The evaluation of the basal activity of cytosolic calcium measured with indo-1 suggested that the disruption of actin or microtubules did not prevent developing muscle cells to maintain a low basal calcium activity. We propose that the global integrity of the cytoskeleton network is not crucial for the maintenance of calcium homeostasis in muscle cells developing in vitro. These results are discussed with regard to current theories attempting to understand the functional consequences of an abnormal expression of the dystrophin-glycoprotein complex interacting with the extracellular matrix and the cytoskeleton.

Animals↗
PubMed1995

[Percutaneous vertebroplasty].

Vertebroplasty is a new therapeutic method which by way of filling with acrylic cement in the vertebral body gives a stabilization of the vertebra and an antalgic effect in painful lesions involving the spine. Main indications consist of spine angiomas, metastases and osteoporotic fractures involving the vertebral body. In most patients, vertebroplasty gives a very good and durable antalgic effect as in cases of angiomas or metastases as in post-therapeutic sequellae in malignant hematosarcomas. In osteoporotic fractures, antalgic effect is obtained very rapidly, but the follow-up is actually insufficient to evaluate the long term benefit.

Hemangioma↗
PubMed1995

Cerebral dural arteriovenous fistulas: clinical and angiographic correlation with a revised classification of venous drainage.

PURPOSE: To review the symptoms and progression of dural arteriovenous fistulas (AVFs) and correlate the findings with various angiographic patterns. MATERIALS AND METHODS: Patterns of venous drainage allowed classification of dural AVFs into five types: type I, located in the main sinus, with antegrade flow; type II, in the main sinus, with reflux into the sinus (IIa), cortical veins (IIb), or both (IIa + b); type III, with direct cortical venous drainage without venous ectasia; type IV, with direct cortical venous drainage with venous ectasia; and type V, with spinal venous drainage. RESULTS: Type I dural AVFs had a benign course. In type II, reflux into the sinus induced intracranial hypertension in 20% of cases, and reflux into cortical veins induced hemorrhage in 10%. Hemorrhage was present in 40% of cases of type III dural AVFs and 65% of type IV. Type V produced progressive myelopathy in 50% of cases. CONCLUSION: This classification provides useful data for determination of the risk with each dural AVF and enables decision-making about the appropriate therapy.

Arteriovenous Fistula↗
PubMed1993

Activation of a slow outward current by the calcium released during contraction of cultured rat skeletal muscle cells.

A slow outward current, activated during depolarization, which induced contraction in whole-cell patch-clamped rat skeletal muscle cells in primary culture [10], was extensively characterized in the present study. This current, Io, was simultaneously recorded with the contraction as a slow outward current during the test pulse, and a slow outward bell-shaped tail after repolarization. Io never appeared below the threshold potential for contraction, and the tail amplitude displayed a similar evolution with peak contraction amplitude as a function of membrane potential. This feature is consistent with the fact that Io was suppressed when contraction was blocked by 5 microM nifedipine [10], and it suggests that Io was dependent on calcium released during contraction. This was confirmed by the fact that the presence of 10 mM EGTA in the patch pipette prevented the development of both contraction and Io, and that Io could be activated during caffeine-induced contractures without applying depolarizations. Io could be carried by K+ or Cs+ ions, but not by Na+. The pharmacology of Io was different from that of Ca(2+)-dependent BK and SK channels, since it was resistant to tetraethylammonium (135 mM), charybdotoxin (25 nM) and apamin (50 nM). Io was also insensitive to 4-aminopyridine (1 mM) but blocked by 5 mM Ba2+ without change to contraction. It was concluded that rat cultured myoballs exhibit a Cs+ permeation through an atypical K+ channel type, which is activated by the calcium released during contraction.

4-Aminopyridine↗
PubMed1993

Intracellular calcium transients induced by different kinds of stimulus during myogenesis of rat skeletal muscle cells studied by laser cytofluorimetry with Indo-1.

Resting intracellular calcium levels and intracellular calcium transients induced by three types of stimulus (acetylcholine, high potassium and caffeine) were recorded, during in vitro myogenesis, by means of a ratiometric fluorescence method using the calcium probe Indo-1 under laser illumination. Resting levels seemed to decrease with the age of cultured cells and the depolarization-induced transients, through 100 mM K+ or Ach application, were progressively faster and larger as the muscle cells developed. An additive mechanism, likely due to calcium entry into the cell through nicotinic acetylcholine receptors, could explain the differences observed in Ach-induced responses as compared with the 100 mM K(+)-induced ones. In myoballs (the older cells) the calcium transients exhibited progressively a biphasic shape. From data obtained in different conditions (tetrodotoxin, nifedipine, strontium and free Ca EGTA) and those indicating the appearance of caffeine-releasable intracellular calcium stores only at 2-3 days stage, and from the previously reported developmental appearance of calcium currents and contraction, it was proposed that, in young myotubes, the calcium transients were more dependent on extracellular calcium than in older cells. These developmental data are discussed in the light of a known model of the in situ biogenesis of the structures involved in excitation-contraction coupling (ECC) like transverse tubules and triads.

Acetylcholine↗
PubMed1993

Changes in cytosolic resting ionized calcium level and in calcium transients during in vitro development of normal and Duchenne muscular dystrophy cultured skeletal muscle measured by laser cytofluorimetry using indo-1.

Intracellular calcium activity was recorded during in vitro myogenesis of human normal and DMD muscle, using the calcium probe Indo-1 under laser illumination, at rest and during different kinds of stimulation (acetylcholine, high K+, caffeine). In myoblasts, the resting intracellular calcium level was significantly larger in DMD cells (89 +/- 9 nM; n = 40 vs 37 +/- 5 nM; n = 22) but there was no significant difference in myotubes, after fusion (44 +/- 4 nM; n = 34 vs 36 +/- 4 nM; n = 52). A similar evolution was observed in cells cultured from FSH biopsies. The amplitude of ACh- and high K(+)-induced calcium transients was significantly halved in DMD myotubes as compared to control ones and non-significantly decreased for caffeine responses. Some alterations in the kinetics of responses were observed in DMD muscle: the rising phases of ACh- and high K(+)-elicited transients and the decaying phase of the ACh-responses were significantly slowed down. It is concluded that: (i) in aneurally cultured human muscle, an increase in the basal level of internal calcium can occur at early stages of myogenesis before the expression of the dystrophin gene; and (ii) the changes in calcium transients induced by depolarization or direct stimulation of sarcoplasmic reticulum are not susceptible of inducing a calcium overload in DMD cells.

Acetylcholine↗