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1.
Sud Med Ekspert ; 66(3): 10-13, 2023.
Artigo em Russo | MEDLINE | ID: mdl-37192452

RESUMO

Aim of this study is to establish the distance and spacing of gunshot wound in the conditions of after-penetration target location. The experimental studies series on shooting coarse colico targets located behind a triplex (car windshield) with record of results by a high-speed camera with a frequency of 1000 frames per second, followed by a study of the affected targets by routine and modern methods (scanning electron microscopy and energy dispersion analysis) were performed. Established results showed that fragments of projectile that overcame an obstacle and secondary projectiles have a rather stable zoning distribution behind an obstacle at a distance up to 2.5 m from the obstacle. The most informative diagnosis of an after-penetration wound distance zone is based on the character of the firearm projectile fragments dispersion, manifestation of lead particles thermal exposure from its sleeve and secondary projectiles fragments formed under the effect of shock wave superimposement in the obstacle material.


Assuntos
Armas de Fogo , Ferimentos por Arma de Fogo , Humanos , Microscopia Eletrônica de Varredura , Balística Forense
2.
Sud Med Ekspert ; 66(1): 39-42, 2023.
Artigo em Russo | MEDLINE | ID: mdl-36719312

RESUMO

The data of the world literature characterizing the forensic aspects of blunt trauma are generalized and systematized. The issues related to the morphology of damage caused after overcoming an obstacle made of various materials by a bullet, the trajectory of a firearm projectile after overcoming an obstacle, the deformation of a bullet, the transfer of fragments of an obstacle by a bullet, the determination of the distance to the target located behind the damaged barrier, the study of the strength properties of individual human armor protection, the nature of the release of particles of an obstacle. The current state of forensic medical examination of blunt trauma is considered, problems and promising directions of their solution are identified. The importance of using knowledge in the field of theoretical mechanics of deformed solid bodies and high-speed impact interaction of solid bodies is noted.


Assuntos
Armas de Fogo , Ferimentos por Arma de Fogo , Ferimentos não Penetrantes , Humanos , Balística Forense , Ferimentos por Arma de Fogo/diagnóstico , Ferimentos não Penetrantes/diagnóstico
3.
Sud Med Ekspert ; 65(6): 21-24, 2022.
Artigo em Russo | MEDLINE | ID: mdl-36472175

RESUMO

The purpose of the work was to study the movement trajectory of the projectile after breaking through an obstacle made of various materials installed at various angles to the aiming line when firing from rifled and traumatic weapons. As a result of a series of experimental shots by shells fired from a traumatic pistol «Jorge¼ (9×22 mm RA cartridge with an initial velocity of 300±20 m/s and a bullet weight of 0.7 g) and the Blaser R93 carbine (Norma Jaktmatch 9.3×62 mm hunting cartridge with an initial speed of 765 m/s and a bullet weight of 11 g) it has been found that, when punching both biological simulators and triplex glass located at the angles of 40-60° with respect to the aiming line, a deflection of the firearm from the aiming line is observed.


Assuntos
Armas de Fogo , Ferimentos por Arma de Fogo , Humanos , Balística Forense/métodos , Ferimentos por Arma de Fogo/etiologia
4.
Sud Med Ekspert ; 65(1): 49-51, 2022.
Artigo em Russo | MEDLINE | ID: mdl-35142472

RESUMO

The article presents a case of fatal gunshot wounds in occupants of a car. The wounds occurred after the passage of gunfire projectiles through a vehicle's triplex glass barrier and had certain features that distinguished them from the common wounds due to the characteristics of the barrier. As part of the solution of the expert questions, the following up-to-date research methods were applied: scanning electron microscopy, energy-dispersive analysis, and computerized three-dimensional reconstruction of the incident circumstances. These research methods provided a clear-cut solution for the situational tasks set by the investigator and served as tools for assessment of the mechanism of bodies injury.


Assuntos
Ferimentos por Arma de Fogo , Automóveis , Humanos , Ferimentos por Arma de Fogo/etiologia
5.
Sud Med Ekspert ; 60(6): 18-20, 2017.
Artigo em Russo | MEDLINE | ID: mdl-29256480

RESUMO

This article was designed to share the experience gained with the three-dimensional modeling for the purpose of situational expertise intended to reconstruct the occurrence circumstances and check up the alternative investigative leads concerning formation of potential injuries to a concrete person. Simulation was performed with the use of the dimensionally scaled model of the place of occurrence as well as the models of the human head and body totally consistent with the anthropometric characteristics of the victim. The results of this work made it possible to reject several potential opportunities for the formation of injuries to the victim and identify the most probable version.


Assuntos
Traumatismos Faciais , Processamento de Imagem Assistida por Computador , Imageamento Tridimensional , Adulto , Antropometria/instrumentação , Antropometria/métodos , Diagnóstico Diferencial , Prova Pericial/métodos , Traumatismos Faciais/diagnóstico , Traumatismos Faciais/diagnóstico por imagem , Medicina Legal/métodos , Humanos , Processamento de Imagem Assistida por Computador/instrumentação , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/instrumentação , Imageamento Tridimensional/métodos , Masculino
6.
Neuroscience ; 131(2): 275-82, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15708472

RESUMO

17Beta-estradiol (E2) is a major neuroregulator, exerting both genomic and non-genomic actions. E2 regulation of Slack (sequence like a calcium-activated potassium channel) potassium channels has not been identified in the CNS. We demonstrate E2-induced activation of Slack channels, which display a unitary conductance of about 60 pS, are inhibited by intracellular calcium, and are abundantly expressed in the nervous system. In lipid bilayers derived from rat cortical neuronal membranes, E2 increases Slack open probability and appears to decrease channel inactivation. Additionally, E2 binds to the Slack channel and activates outward currents in human embryonic kidney-293 cells that express Slack channels but not classical estrogen receptors (i.e. ERalpha or ERbeta). Neither E2-induced activation nor the binding intensity of E2 to the Slack channel is blocked by tamoxifen, an ER antagonist/agonist. Thus, E2 activates a potassium channel, Slack, through a non-traditional membrane binding site, adding to known non-genomic mechanisms by which E2 exerts pharmacological and toxicological effects in the CNS.


Assuntos
Estradiol/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Canais de Potássio/metabolismo , Animais , Linhagem Celular , Relação Dose-Resposta a Droga , Humanos , Proteínas do Tecido Nervoso/genética , Neurônios/metabolismo , Canais de Potássio/genética , Canais de Potássio Ativados por Sódio , Ligação Proteica/fisiologia
7.
Synapse ; 37(1): 38-55, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10842350

RESUMO

Exocytosis is commonly viewed as the only secretory process able to account for quantal forms of fast synaptic transmission. However, the demonstrated variability and composite properties of miniature postsynaptic signals are not easily explained by all-or-none exocytotic discharge of transmitter in solution from inside vesicles. Recent studies of endocrine secretion have shown that hormone release does not coincide with exocytosis due to its trapping in the core matrix of the granule. Thus, we tested whether the synaptic transmitter GABA could also be held in a matrix before being released. Using confocal microscopy and flow cytometry of embryonic rat hippocampal neurons, we found a GABA immunoreaction at the surface of live cell bodies and growth cones that coincided spatially and quantitatively with the binding of tetanus toxin fragment C (TTFC). TTFC binds predominantly at membrane sites containing the trisialoglycosphingolipid GT1b. Using flow cytometry, GT1b-containing liposomes preincubated in 100 nM GABA exhibited the same relationship between GABA and TTFC surface binding as found on neurons and growth cones. Embryonic neurons differentiated in culture expressed initially a tonic, and after 3-5 days, transient, postsynaptic signals mediated by GABA acting at GABA(A) receptor/Cl(-) channels. A stream of saline applied to the neuronal surface rapidly and reversibly suppressed both tonic and transient signals. A brief application of the GABAmimetic isoguvacine immediately transformed both tonic and transient GABAergic signals into tonic and transient isoguvacinergic signals. These results and those in the literature are consistent with an immediately releasable compartment of transmitter accessible from the presynaptic surface.


Assuntos
Exocitose/fisiologia , Neurônios/metabolismo , Transmissão Sináptica/fisiologia , Ácido gama-Aminobutírico/metabolismo , Adsorção , Animais , Células Cultivadas , Canais de Cloreto/metabolismo , Eletrofisiologia , Feminino , Citometria de Fluxo , Agonistas GABAérgicos/farmacologia , Gangliosídeos/análise , Ácidos Isonicotínicos/farmacologia , Lipossomos/química , Lipossomos/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Proteínas de Membrana/análise , Proteínas de Membrana/metabolismo , Neurônios/química , Neurônios/citologia , Gravidez , Terminações Pré-Sinápticas/química , Terminações Pré-Sinápticas/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de GABA-A/metabolismo , Ácido gama-Aminobutírico/análise
8.
Vestn Otorinolaringol ; (6): 24-6, 2000.
Artigo em Russo | MEDLINE | ID: mdl-11187070

RESUMO

Pharmacopuncture treatment (cerebrolysin, cavinton, prozerin) was given to 120 children with neurosensory hypoacusis aged 4-15 years. The treatment consisted of 3 courses 10 days each with 7-10-day interval. The drugs were injected in the dose 0.1 ml in biologically active points according to the original technique. The treatment lowered tonal thresholds more than by 15 dB in 36% children. Speech intelligibility increased, head ache and nose in the ears relieved in 85% cases. Adaptation to the hearing aid was positive. Clinical evidence was confirmed by REG picture indicating normalization of hemodynamics. Pharmacopuncture is a promising method in the treatment of children with neurosensory hypoacusis.


Assuntos
Terapia por Acupuntura/métodos , Perda Auditiva Central/tratamento farmacológico , Nootrópicos/administração & dosagem , Parassimpatomiméticos/administração & dosagem , Extratos de Tecidos/administração & dosagem , Adolescente , Aminoácidos/administração & dosagem , Limiar Auditivo/efeitos dos fármacos , Encéfalo , Criança , Pré-Escolar , Quimioterapia Combinada , Feminino , Perda Auditiva Central/fisiopatologia , Humanos , Injeções Intradérmicas , Masculino , Neostigmina/administração & dosagem , Prognóstico , Alcaloides de Vinca/administração & dosagem
9.
Biophys J ; 76(4): 1744-56, 1999 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10096875

RESUMO

The dihydropyridine receptor (DHPR) of skeletal muscle functions as a Ca2+ channel and is required for excitation-contraction (EC) coupling. Here we show that the DHPR beta subunit is involved in the regulation of these two functions. Experiments were performed in skeletal mouse myotubes selectively lacking a functional DHPR beta subunit. These beta-null cells have a low-density L-type current, a low density of charge movements, and lack EC coupling. Transfection of beta-null cells with cDNAs encoding for either the homologous beta1a subunit or the cardiac- and brain-specific beta2a subunit fully restored the L-type Ca2+ current (161 +/- 17 pS/pF and 139 +/- 9 pS/pF, respectively, in 10 mM Ca2+). We compared the Boltzmann parameters of the Ca2+ conductance restored by beta1a and beta2a, the kinetics of activation of the Ca2+ current, and the single channel parameters estimated by ensemble variance analysis and found them to be indistinguishable. In contrast, the maximum density of charge movements in cells expressing beta2a was significantly lower than in cells expressing beta1a (2.7 +/- 0.2 nC/microF and 6.7 +/- 0. 4 nC/microF, respectively). Furthermore, the amplitude of Ca2+ transient measured by confocal line-scans of fluo-3 fluorescence in voltage-clamped cells were 3- to 5-fold lower in myotubes expressing beta2a. In summary, DHPR complexes that included beta2a or beta1a restored L-type Ca2+ channels. However, a DHPR complex with beta1a was required for complete restoration of charge movements and skeletal-type EC coupling. These results suggest that the beta1a subunit participates in key regulatory events required for the EC coupling function of the DHPR.


Assuntos
Canais de Cálcio/química , Canais de Cálcio/metabolismo , Cálcio/metabolismo , Músculo Esquelético/metabolismo , Animais , Fenômenos Biofísicos , Biofísica , Canais de Cálcio/genética , Canais de Cálcio Tipo L , Células Cultivadas , DNA Complementar/genética , Transporte de Íons , Cinética , Potenciais da Membrana , Camundongos , Contração Muscular/fisiologia , Conformação Proteica , Coelhos , Transfecção
10.
Biophys J ; 75(1): 207-17, 1998 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9649380

RESUMO

The origin of Ibetanull, the Ca2+ current of myotubes from mice lacking the skeletal dihydropyridine receptor (DHPR) beta1a subunit, was investigated. The density of Ibetanull was similar to that of Idys, the Ca2+ current of myotubes from dysgenic mice lacking the skeletal DHPR alpha1S subunit (-0.6 +/- 0.1 and -0.7 +/- 0.1 pA/pF, respectively). However, Ibetanull activated at significantly more positive potentials. The midpoints of the GCa-V curves were 16.3 +/- 1.1 mV and 11.7 +/- 1.0 mV for Ibetanull and Idys, respectively. Ibetanull activated significantly more slowly than Idys. At +30 mV, the activation time constant for Ibetanull was 26 +/- 3 ms, and that for Idys was 7 +/- 1 ms. The unitary current of normal L-type and beta1-null Ca2+ channels estimated from the mean variance relationship at +20 mV in 10 mM external Ca2+ was 22 +/- 4 fA and 43 +/- 7 fA, respectively. Both values were significantly smaller than the single-channel current estimated for dysgenic Ca2+ channels, which was 84 +/- 9 fA under the same conditions. Ibetanull and Idys have different gating and permeation characteristics, suggesting that the bulk of the DHPR alpha1 subunits underlying these currents are different. Ibetanull is suggested to originate primarily from Ca2+ channels with a DHPR alpha1S subunit. Dysgenic Ca2+ channels may be a minor component of this current. The expression of DHPR alpha1S in beta1-null myotubes and its absence in dysgenic myotubes was confirmed by immunofluorescence labeling of cells.


Assuntos
Canais de Cálcio/deficiência , Cálcio/metabolismo , Músculo Esquelético/metabolismo , Animais , Fenômenos Biofísicos , Biofísica , Canais de Cálcio/química , Canais de Cálcio/genética , Canais de Cálcio Tipo L , Células Cultivadas , Ativação do Canal Iônico , Cinética , Potenciais da Membrana , Camundongos , Mutação
11.
Biophys J ; 73(2): 807-18, 1997 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-9251797

RESUMO

The Ca2+ currents, charge movements, and intracellular Ca2+ transients of mouse dihydropyridine receptor (DHPR) beta 1-null myotubes expressing a mouse DHPR beta 1 cDNA have been characterized. In beta 1-null myotubes maintained in culture for 10-15 days, the density of the L-type current was approximately 7-fold lower than in normal cells of the same age (Imax was 0.65 +/- 0.05 pA/pF in mutant versus 4.5 +/- 0.8 pA/pF in normal), activation of the L-type current was significantly faster (tau activation at +40 mV was 28 +/- 7 ms in mutant versus 57 +/- 8 ms in normal), charge movements were approximately 2.5-fold lower (Qmax was 2.5 +/- 0.2 nC/microF in mutant versus 6.3 +/- 0.7 nC/microF in normal), Ca2+ transients were not elicited by depolarization, and spontaneous or evoked contractions were absent. Transfection of beta 1-null cells by lipofection with beta 1 cDNA reestablished spontaneous or evoked contractions in approximately 10% of cells after 6 days and approximately 30% of cells after 13 days. In contracting beta 1-transfected myotubes there was a complete recovery of the L-type current density (Imax was 4 +/- 0.9 pA/pF), the kinetics of activation (tau activation at +40 mV was 64 +/- 5 ms), the magnitude of charge movements (Qmax was 6.7 +/- 0.4 nC/microF), and the amplitude and voltage dependence of Ca2+ transients evoked by depolarizations. Ca2+ transients of transfected cells were unaltered by the removal of external Ca2+ or by the block of the L-type Ca2+ current, demonstrating that a skeletal-type excitation-contraction coupling was restored. The recovery of the normal skeletal muscle phenotype in beta 1-transfected beta-null myotubes shows that the beta 1 subunit is essential for the functional expression of the DHPR complex.


Assuntos
Canais de Cálcio/deficiência , Canais de Cálcio/fisiologia , Cálcio/metabolismo , Músculo Esquelético/fisiologia , Animais , Canais de Cálcio/biossíntese , Canais de Cálcio Tipo L , Cloreto de Cálcio/farmacologia , Células Cultivadas , Condutividade Elétrica , Feto , Substâncias Macromoleculares , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Camundongos Knockout , Contração Muscular , Músculo Esquelético/citologia , Músculo Esquelético/embriologia , Técnicas de Patch-Clamp , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Transfecção
12.
J Membr Biol ; 154(1): 81-9, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8881029

RESUMO

There is increasing evidence that Ca2+ release from sarcoplasmic reticulum (SR) of mammalian skeletal muscle is regulated or modified by several factors including ionic composition of the myoplasm. We have studied the effect of Cl- on the release of Ca2+ from the SR of rabbit skeletal muscle in both skinned psoas fibers and in isolated terminal cisternae vesicles. Ca2+ release from the SR in skinned fibers was inferred from increases in isometric tension and the amount of release was assessed by integrating the area under each tension transient. Ca2+ release from isolated SR was measured by rapid filtration of vesicles passively loaded with 45Ca2+. Ca2+ release from SR was stimulated in both preparations by exposure to a solution containing 191 mm choline-Cl, following pre-equilibration in Ca2+-loading solution that had propionate as the major anion. Controls using saponin (50 microg/ml), indicated that the release of Ca2+ was due to direct action of Cl- on the SR rather than via depolarization of T-tubules. Procaine (10 mM) totally blocked Cl-- and caffeine-elicited tension transients recorded using loading and release solutions having ([Na+] + [K+]) x [Cl-] product of 6487.69 mm2 and 12361.52 mm2, respectively, and blocked 60% of Ca2+ release in isolated SR vesicles. Surprisingly, procaine had only a minor effect on tension transients elicited by Cl- and caffeine together. The data from both preparations suggests that Cl- induces a relatively small amount of Ca2+ release from the SR by activating receptors other than RYR-1. In addition, Cl- may increase the Ca2+ sensitivity of RYR-1, which would then allow the small initial release of Ca2+ to facilitate further release of Ca2+ from the SR by Ca2+-induced Ca2+ release.


Assuntos
Cálcio/metabolismo , Colina/farmacologia , Músculos Psoas/metabolismo , Retículo Sarcoplasmático/metabolismo , Animais , Músculos Psoas/ultraestrutura , Coelhos
13.
Proc Natl Acad Sci U S A ; 93(24): 13961-6, 1996 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-8943043

RESUMO

The multisubunit (alpha 1s, alpha 2/delta, beta 1, and gamma) skeletal muscle dihydropyridine receptor transduces transverse tubule membrane depolarization into release of Ca2+ from the sarcoplasmic reticulum, and also acts as an L-type Ca2+ channel. The alpha 1s subunit contains the voltage sensor and channel pore, the kinetics of which are modified by the other subunits. To determine the role of the beta 1 subunit in channel activity and excitation-contraction coupling we have used gene targeting to inactivate the beta 1 gene. beta 1-null mice die at birth from asphyxia. Electrical stimulation of beta 1-null muscle fails to induce twitches, however, contractures are induced by caffeine. In isolated beta 1-null myotubes, action potentials are normal, but fail to elicit a Ca2+ transient. L-type Ca2+ current is decreased 10- to 20-fold in the beta 1-null cells compared with littermate controls. Immunohistochemistry of cultured myotubes shows that not only is the beta 1 subunit absent, but the amount of alpha 1s in the membrane also is undetectable. In contrast, the beta 1 subunit is localized appropriately in dysgenic, mdg/mdg, (alpha 1s-null) cells. Therefore, the beta 1 subunit may not only play an important role in the transport/insertion of the alpha 1s subunit into the membrane, but may be vital for the targeting of the muscle dihydropyridine receptor complex to the transverse tubule/sarcoplasmic reticulum junction.


Assuntos
Canais de Cálcio/biossíntese , Canais de Cálcio/genética , Cálcio/metabolismo , Contração Muscular , Músculo Esquelético/fisiologia , Potenciais de Ação/fisiologia , Animais , Canais de Cálcio/fisiologia , Canais de Cálcio Tipo L , Clonagem Molecular , Embrião de Mamíferos , Biblioteca Genômica , Substâncias Macromoleculares , Camundongos , Camundongos Knockout , Miofibrilas/fisiologia , Miofibrilas/ultraestrutura , Proteínas Recombinantes/biossíntese , Retículo Sarcoplasmático/metabolismo
14.
Biophys J ; 71(2): 707-21, 1996 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-8842209

RESUMO

We purified and characterized ryanotoxin, an approximately 11.4-kDa peptide from the venom of the scorpion Buthotus judiacus that induces changes in ryanodine receptors of rabbit skeletal muscle sarcoplasmic reticulum analogous to those induced by the alkaloid ryanodine. Ryanotoxin stimulated Ca2+ release from sarcoplasmic reticulum vesicles and induced a state of reduce unit conductance with a mean duration longer than that of unmodified ryanodine receptor channels. With Cs+ as the current carrier, the slope conductance of the state induced by 1 microM ryanotoxin was 163 +/- 12 pS, that of the state induced by 1 microM ryanodine was 173 +/- 26 pS, and that of control channels was 2.3-fold larger (396 +/- 25 pS). The distribution of substate events induced by 1 microM RyTx was biexponential and was fitted with time constants approximately 10 times shorter than those fitted to the distribution of substates induced by 1 microM ryanodine. Bath-applied 5 microM ryanotoxin had no effect on the excitability of mouse myotubes in culture. When 5 microM ryanotoxin was dialyzed into the cell through the patch pipette in the whole-cell configuration, there was a voltage-dependent increase in the amplitude of intracellular Ca2+ transients elicited by depolarizing potentials in the range of -30 to +50 mV. Ryanotoxin increased the binding affinity of [3H]ryanodine in a reversible manner with a 50% effective dose (ED50) of 0.16 microM without altering the maximum number (Bmax) of [3H]ryanodine-binding sites. This result suggested that binding sites for ryanotoxin and ryanodine were different. Ryanotoxin should prove useful in identifying domains coupling the ryanodine receptor to the voltage sensor, or domains affecting the gating and conductance of the ryanodine receptor channel.


Assuntos
Canais de Cálcio/fisiologia , Cálcio/metabolismo , Proteínas Musculares/fisiologia , Músculo Esquelético/fisiologia , Rianodina/farmacologia , Retículo Sarcoplasmático/fisiologia , Venenos de Escorpião , Venenos de Escorpião/farmacologia , Animais , Canais de Cálcio/efeitos dos fármacos , Células Cultivadas , Césio/farmacologia , Cromatografia Líquida de Alta Pressão , Condutividade Elétrica , Feto , Cinética , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Proteínas Musculares/efeitos dos fármacos , Coelhos , Rutênio Vermelho/farmacologia , Rianodina/metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina , Venenos de Escorpião/isolamento & purificação
15.
FEBS Lett ; 352(3): 285-90, 1994 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-7925988

RESUMO

45Ca2+ fluxes and planar bilayer recordings indicated that the fatty acid metabolite palmitoyl coenzyme A, but not free coenzyme A or palmitic acid, stimulated the cardiac ryanodine receptor channel of pig heart sarcoplasmic reticulum. Palmitoyl CoA reactivated channels inhibited by concentrations of cytoplasmic free Mg2+ in the physiological range. Reactivation by palmitoyl CoA in the presence of Mg2+ was stimulated by myoplasmic free Ca2+ in the micromolar range. Acyl coenzyme A derivatives may be utilized by cardiac muscle cells to compensate for the severe Mg2+ inhibition of ryanodine receptors which would otherwise leave Ca2+ stores unresponsive to Ca2+ and to other cytosolic ligands involved in signal transduction.


Assuntos
Canais de Cálcio/fisiologia , Cálcio/metabolismo , Magnésio/farmacologia , Proteínas Musculares/fisiologia , Miocárdio/metabolismo , Palmitoil Coenzima A/farmacologia , Retículo Sarcoplasmático/metabolismo , Animais , Cálcio/farmacologia , Canais de Cálcio/efeitos dos fármacos , Ventrículos do Coração , Cinética , Bicamadas Lipídicas , Proteínas Musculares/efeitos dos fármacos , Canal de Liberação de Cálcio do Receptor de Rianodina , Suínos , Fatores de Tempo
16.
Biophys J ; 67(2): 751-65, 1994 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-7948689

RESUMO

We investigated the effect of Cl- on the Ca2+ permeability of rabbit skeletal muscle junctional sarcoplasmic reticulum (SR) using 45Ca2+ fluxes and single channel recordings. In 45Ca2+ efflux experiments, the lumen of the SR was passively loaded with solutions of 150 mM univalent salt containing 5 mM 45Ca2+. Release of 45Ca2+ was measured by rapid filtration in the presence of extravesicular 0.4-0.8 microM free Ca2+ and 150 mM of the same univalent salt loaded into the SR lumen. The rate of release was 5-10 times higher when the univalent salt equilibrated across the SR-contained Cl- (Tris-Cl, choline-Cl, KCl) instead of an organic anion or other halides (gluconate-, methanesulfonate-, acetate-, HEPES-, Br-, I-). Cations (K+, Tris+) could be interchanged without a significant effect on the release rate. To determine whether Cl- stimulated ryanodine receptors, we measured the stimulation of release by ATP (5 mM total) and caffeine (20 mM total) and the inhibition by Mg2+ (0.8 mM estimated free) in Cl(-)-free and Cl(-)-containing solutions. The effects of ATP, caffeine, and Mg2+ were the largest in K-gluconate and Tris-gluconate, intermediate in KCl, and notably poor or absent in choline-Cl and Tris-Cl. Procaine (10 mM) inhibited the caffeine-stimulated release measured in K-gluconate, whereas the Cl- channel blocker clofibric acid (10 mM) but not procaine inhibited the caffeine-insensitive release measured in choline-Cl. Ruthenium red (20 microM) inhibited release in all solutions. In SR fused to planar bilayers we identified a nonselective Cl- channel (PCl: PTris: PCa = 1:0.5:0.3) blocked by ruthenium red and clofibric acid but not by procaine. These conductive and pharmacological properties suggested the channel was likely to mediate Cl(-)-dependent SR Ca2+ release. The absence of a contribution of ryanodine receptors to the Cl(-)-dependent release were indicated by the lack of an effect of Cl- on the open probability of this channel, a complete block by procaine, and a stimulation rather than inhibition by clofibric acid. A plug model of Cl(-)-dependent release, whereby Cl- removed the inhibition of the nonselective channel by large anions, was formulated under the assumption that nonselective channels and ryanodine receptor channels operated separately from each other in the terminal cisternae. The remarkably large contribution of Cl- to the SR Ca2+ permeability suggested that nonselective Cl- channels may control the Ca2+ permeability of the SR in the resting muscle cell.


Assuntos
Canais de Cálcio/fisiologia , Cálcio/metabolismo , Cloretos/farmacologia , Proteínas Musculares/fisiologia , Músculos/fisiologia , Retículo Sarcoplasmático/fisiologia , Trifosfato de Adenosina/metabolismo , Animais , Ânions/farmacologia , Cafeína/farmacologia , Canais de Cálcio/efeitos dos fármacos , Radioisótopos de Cálcio , Clofibrato/farmacologia , Ativação do Canal Iônico , Cinética , Bicamadas Lipídicas , Potenciais da Membrana/fisiologia , Modelos Biológicos , Proteínas Musculares/efeitos dos fármacos , Procaína/farmacologia , Coelhos , Rianodina/metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina , Retículo Sarcoplasmático/efeitos dos fármacos , Fatores de Tempo
17.
Am J Physiol ; 266(6 Pt 1): C1485-504, 1994 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-8023884

RESUMO

Membrane depolarization, neurotransmitters, and hormones evoke a release of Ca2+ from intracellular Ca(2+)-storing organelles like the endoplasmic reticulum and, in muscle, the sarcoplasmic reticulum (SR). In turn, the released Ca2+ serves to trigger a variety of cellular responses. The presence of Ca2+ pumps to replenish intracellular stores was described more than 20 years ago. The presence of Ca2+ channels, like the ryanodine receptor, which suddenly release the organelle-stored Ca2+, is a more recent finding. This review describes the progress made in the last five years on the structure, function, and regulation of the ryanodine receptor. Numerous reports have described the response of ryanodine receptors to cellular ions and metabolites, kinases and other proteins, and pharmacological agents. In many cases, comparative measurements have been made using Ca2+ fluxes in SR vesicles, single-channel recordings in planar bilayers, and radioligand binding assays using [3H]ryanodine. These techniques have helped to relate the activity of single ryanodine receptors to global changes in the SR Ca2+ permeability. Molecular information on functional domains within the primary structure of the ryanodine receptor is also available. There are at least three ryanodine receptor isoforms in various tissues. Some cells, such as amphibian muscle cells, express more than a single isoform. The diversity of ligands known to modulate gating and the diversity of tissues known to express the protein suggest that the ryanodine receptor has the potential to participate in many types of cell stimulus-Ca(2+)-release coupling mechanisms.


Assuntos
Canais de Cálcio/química , Canais de Cálcio/metabolismo , Proteínas Musculares/química , Proteínas Musculares/metabolismo , Animais , Cálcio/metabolismo , Canais de Cálcio/isolamento & purificação , Clonagem Molecular , Humanos , Ativação do Canal Iônico , Proteínas Musculares/isolamento & purificação , Fosforilação , Canal de Liberação de Cálcio do Receptor de Rianodina , Retículo Sarcoplasmático/metabolismo , Relação Estrutura-Atividade , Distribuição Tecidual
18.
Biochem Mol Biol Int ; 32(3): 441-8, 1994 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8032313

RESUMO

L-thyroxine activated the Ca2+ release channel (ryanodine receptor) of skeletal muscle. [3H]ryanodine binding was stimulated by L-thyroxine in a dose dependent manner producing a two-fold increase at 250 microM. The same concentration induced a release of approximately 40% of the 45Ca2+ passively loaded into sarcoplasmic reticulum in 100 msec. Ca2+ release channel activity monitored in planar bilayers increased in the presence of 250 microM L-thyroxine from a control open probability of 0.02 +/- 0.03 to 0.17 +/- 0.12. Thyroid hormones may directly open Ca2+ release channels of skeletal muscle, thus altering intracellular Ca2+ homeostasis.


Assuntos
Canais de Cálcio/fisiologia , Cálcio/metabolismo , Proteínas Musculares/fisiologia , Músculos/metabolismo , Retículo Sarcoplasmático/metabolismo , Tiroxina/farmacologia , Animais , Canais de Cálcio/efeitos dos fármacos , Cinética , Bicamadas Lipídicas , Potenciais da Membrana/efeitos dos fármacos , Proteínas Musculares/efeitos dos fármacos , Rianodina/metabolismo , Canal de Liberação de Cálcio do Receptor de Rianodina , Suínos
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