Retinoic Acid and VEGF Delay Smooth Muscle Relative to Endothelial Differentiation to Coordinate Inner and Outer Coronary Vessel Wall Morphogenesis

Rationale: Major coronary vessels derive from the proepicardium, the cellular progenitor of the epicardium, coronary endothelium, and coronary smooth muscle cells (CoSMCs). CoSMCs are delayed in their differentiation relative to coronary endothelial cells (CoEs), such that CoSMCs mature only after CoEs have assembled into tubes. The mechanisms underlying this sequential CoE/CoSMC differentiation are unknown. Retinoic acid (RA) is crucial for vascular development and the main RA-synthesizing enzyme is progressively lost from epicardially derived cells as they differentiate into blood vessel types. In parallel, myocardial vascular endothelial growth factor (VEGF) expression also decreases along coronary vessel muscularization. Objective: We hypothesized that RA and VEGF act coordinately as physiological brakes to CoSMC differentiation. Methods and Results: In vitro assays (proepicardial cultures, cocultures, and RALDH2 [retinaldehyde dehydrogenase-2]/VEGF adenoviral overexpression) and in vivo inhibition of RA synthesis show that RA and VEGF act as repressors of CoSMC differentiation, whereas VEGF biases epicardially derived cell differentiation toward the endothelial phenotype. Conclusion: Experiments support a model in which early high levels of RA and VEGF prevent CoSMC differentiation from epicardially derived cells before RA and VEGF levels decline as an extensive endothelial network is established. We suggest this physiological delay guarantees the formation of a complex, hierarchical, tree of coronary vessels. (Circ Res. 2010;107:204-216.)

Mode of cembranoid action on embryonic muscle acetylcholine receptor

The mechanism of eupalmerin acetate (EUAC) actions on the embryonic muscle nicotinic acetylcholine receptor (nAChR) in BC3H-1 cells was studied by using whole-cell and single-channel patch-clamp current measurements. With whole-cell currents, EUAC did not act as an agonist on this receptor. Coapplication of 30 mu M EUAC with 50 mu M, 100 N, or 500 mu M carbamoylcholine (CCh) reversibly inhibited the current amplitude, whereas, with 20 mu M CCh, current was increased above control values in the presence of EUAC. EUAC concentration curves (0.01-40 N) obtained with 100 mu M and 500 mu M CCh displayed slope coefficients, n(H), significantly smaller than one, suggesting that EUAC bound to several sites with widely differing affinities on the receptor molecule. The apparent rate of receptor desensitization in the presence of EUAC and CCh was either slower than or equal to that obtained with CCh alone. The major finding from single-channel studies was that EUAC did not affect single-channel conductance or the ability of CCh to interact with the receptor. Instead, EUAC acted by increasing the channel closing rate constant. The results are not consistent with the competitive model for EUAC inhibition, with the sequential open-channel block model, or with inhibition by increased desensitization. The data are best accounted for by a model in which EUAC acts by closed-channel block at low concentrations, by positive modulation at intermediate concentrations, and by negative allosteric modulation of the open channel at high concentrations. (c) 2007 Wiley-Liss, Inc.

Inhibition of the myotoxic activity of Bothrops jararacussu venom and its two major myotoxins, BthTX-I and BthTX-II, by the aqueous extract of Tabernaemontana catharinensis A. DC. (Apocynaceae)

Partial neutralization of the myotoxic effect of Bothrops jararacussu venom (BV) and two of its myotoxins [bothropstoxin-I (BthTX-I), catalytically inactive, and II (BthTX-II), showing low PLA(2) activity], by the lyophilized aqueous extract of Tabernaemontana catharinensis (AE), was studied in rat isolated soleus muscle preparations (in vitro) and through i.m. injection in the gastrocnemius muscle (in vivo) by determination of creatine kinase (CK) activity and histopathological analysis. Incubation of soleus muscle for 1 h with BV or toxins (20 mug/ml) plus AE (400 mug/ml) added immediately after BV, BthTX-I or BthTX-II reduced CK levels by 53%, 37% and 56%, respectively. The myonecrotic effects of BV (20 mug/ml) upon soleus muscle was reduced 24%, 35% and 36% when AE (400 mug/ml) was added 1 h after BV and CK was evaluated 30 min, 1 and 2 h later, respectively. For BthTX-I these values were 46%, 48% and 47%, while for BthTX-II no inhibitory effect was detected. Histological analysis of soleus muscle after incubation with AE (400 mug/ml, I h) did not reveal any change in muscle fibers, but severe necrosis induced by -BV or toxins (20 mug/ml) was clearly in evidence, and decreased significantly when soleus muscle was protected by AE. This protection was also observed when AE was administered 1 h after BV or BthTX-I, but not after BthTX-II. AE did not inhibit the catalytic PLA(2), activity of BthTX-II or BV and did not change the PAGE pattern of BV, BthTX-I or BthTX-II. In vivo assays were performed in 100-g rats and maximal CK release was attained at a dose of 100 mug of BV, 3 h after injection. AE was not effective when injected 20 s after BV or toxins. However, injecting BV or toxins (100 mug), which were pre-incubated with AE (2 mg) caused an inhibition of 57%, 59% and 51%, respectively, with zero time pre-incubation, but was less effective with I h pre-incubation. This plant represents a potential source of promising myotoxin inhibitors. (C) 2004 Elsevier GmbH. All rights reserved.

Maternal protein restriction induce skeletal muscle changes without altering the MRFs MyoD and myogenin expression in offspring

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological aspects of neuromuscular junctions and gene expression of nicotinic acetylcholine receptors (nAChRs) in skeletal muscle of rats with heart failure

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Mastoparan effects in skeletal muscle damage: An ultrastructural view until now concealed

Animal venoms have been valuable sources for development of new drugs and important tools to understand cellular functioning in health and disease. The venom of Polybia paulista, a neotropical social wasp belonging to the subfamily Polistinae, has been sampled by headspace solid phase microextraction and analyzed by gas chromatography-mass spectrometry. Recent study has shown that mastoparan, a major basic peptide isolated from the venom, reproduces the myotoxic effect of the whole venom. In this study, Polybia-MPII mastoparan was synthesized and studies using transmission electron microscopy were carried out in mice tibial anterior muscle to identify the subcellular targets of its myotoxic action. The effects were followed at 3 and 24 h, 3, 7, and 21 days after mastoparan (0.25 mu g/mu L) intramuscular injection. The peptide caused disruption of the sarcolemma and collapse of myofibril arrangement in myofibers. As a consequence, fibers presented heteromorphic amorphous masses of agglutinated myofilaments very often intermingled with denuded sarcoplasmic areas sometimes only surrounded by a persistent basal lamina. To a lesser extent, a number of fibers apparently did not present sarcolemma rupture but instead appeared with multiple small vacuoles. The results showed that sarcolemma, sarcoplasmic reticulum (SR), and mitochondria were the main targets for mastoparan. In addition, a number of fibers showed apoptotic-like nuclei suggesting that the peptide causes death both by necrosis and apoptosis. This study presents a hitherto unexplored view of the effects of mastoparan in skeletal muscle and contributes to discuss how the known pharmacology of the peptide is reflected in the sarcolemma, SR, mitochondria, and nucleus of muscle fibers, apparently its subcellular targets.

Electromyographic study of levator scapulae and rhomboideus major muscles in movements of the shoulder and arm

In 21 normal adult male subjects, the muscular activity of the levator scapulae and rhomboideus major muscles was studied electromyographically during the movements of the shoulder and arm. Two single coaxial needle electrodes were used for registering the action potentials. Concerning shoulder movement, it was shown that the levator scapulae was active in elevation and rhomboideus major was active in retraction. Both muscles were inactive during protrusion, in most events. Concerning free movements of the arm, both muscles were active in abduction, elevation, adduction, flexion and circumduction, but inactive in extension. During the same movements, performed with a load, we observed greater intensity in the activity of these muscles in comparison to their activity during free movements.

Electromyographic study of the trapezius (middle portion) and rhomboideus major muscles in movements of the trunk

The authors studied the trapezius (middle portion) and rhomboideus major muscles in movements of flexion, extension, inclination and rotation of the trunk. The electromyographic records demonstrate that such muscles show activity only at the ending of flexion, being inactive in the other movements.

Analyse electromyographique des muscles levator scapulae et rhomboideus major dans les mouvements du membre superieur

The participation of the levator scapulae and rhomboideus major muscles in some movements of the upper limb was analysed in 21 young adult male volunteers. A 2 channel TECA TE4 electromyograph connected with single coaxial needle electrodes was used. In abduction, elevation, adduction, flexion and circumduction participation of both muscles in free movements of the upper limb was found. In extension, however, these muscles were inactive. In the same movements analysed with load, a major intensity of action of these muscles was registered as compared with those obtained in free movements.

Electromyographic study of the trapezius (pars media) and rhomboideus major muscles during respiration

The authors studied the trapezius (pars media) and rhomboideus major muscles during deep inspiration and expiration. The electromyographic records demonstrated that these muscles showed no activity in either phase of breathing.

Electromyographic study of the trapezius (pars media) and rhomboideus major muscles in free movements of the head

The authors studied the trapezius (pars media) and rhomboideus major muscles in movements of flexion, extension, inclination and rotation of the head. The electromyographic records demonstrated that referred muscles are inactive in these different movements.

Etude electromyographique des muscles trapezius (partie moyenne) et rhomboideus major sousmis a tension isometrique

The participation of the trapezius (middle portion) and rhomboideus major muscles submitted to an isometric tension (holding downward dumbells of 11, 15 and 19 kg) was analysed in 40 young adult male vounteers. A 2 channel TECA TE4 electromyograph connected with single coaxial needle electrodes was used. In the initial phase of the test, holding downward, generally the trapezius and rhomboideus major muscles showed activity and no activity, respectively. In the cases where activity was present during the downward positions it was reduced gradually until complete rest.

Electromyographic study of the trapezius (pars media) and rhomboideus major muscles in movements of the arm (part 1)

The participation of the trapezius (pars media) and rhomboideus major muscles in free movements of abduction, adduction, flexion, extension and hyperextension of the arm was studied electromyographically. These muscles were active and synergic in all analysed movements, both acting with intensity that ranged from moderate to very marked.

Electromyographic study of the trapezius (pars media) and rhomboideus major muscles in movements of the arm (part 2)

The participation of the trapezius (pars media) and rhomboideus major muscles were studied electromyographically in movements of lateral and medial rotation (free and against resistance) of the arm. In the majority of cases both muscles were inactive during the performance of the analysed movements. Cases in which only one or both muscles acted in free rotation occurred only at the second half or ending of the movement. Records of electrical potentials during against resistance rotation were due to tensional efforts at the shoulder level.

Evolution and genomic organization of muscle microRNAs in fish genomes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)