Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.24.3 (collagenase)
 18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

1. Electrical and contractile properties of resealed fibre segments were investigated by a variety of in vitro techniques. The preparations were removed from skeletal muscles of normal subjects and of eight patients with myotonic dystrophy. 2. Several hours after removal, fibre segments from normal subjects and those patients in whom myotonia was the primary symptom had resting membrane potentials of approximately -80 mV. In contrast, fibre segments obtained from patients in whom muscle dystrophy was more expressed were depolarized (-60 to -70 mV). 3. Contractions induced in fibre segments of myotonic muscle which had normal potentials were characterized by slowed relaxation which was due to electrical after-activity. 4. After single stimuli, long-lasting (3-100) runs of action potentials were recorded intracellularly from the myotonic muscle. In some of these fibre segments complex repetitive discharges were observed: multiple sites of locally gated currents were identified. 5. The three-electrode voltage clamp was used to determine the total membrane conductance, gm, and the ion component conductances. All fibres of a particular patient had similar conductances. However, the Cl- conductance varied from patient to patient from normal (74% of gm) to low values (30% of gm). The K+ conductance was normal in all fibres of all patients. 6. The patch-clamp technique was used to record currents through single Na+ channels of the sarcolemma. After treatment of the fibre segments with collagenase gigaohm seals were routinely obtained. The rate of success was greater when using the cell-attached mode than the inside-out mode. 7. Sodium channel currents were elicited by depolarizing voltage steps which produced an initial burst of Na+ channel openings. Up to ten channels were activated simultaneously when the patch was depolarized to potentials more positive than -30 mV. The Na+ channels re-opened very rarely in controls. The macroscopic sodium current, INa, was reconstructed by averaging depolarizing pulses. The time constant of rapid decay of INa reflecting macroscopic inactivation, the onset of INa and the amplitude of INa were voltage dependent. The mean amplitude of the current produced by re-openings was on average only 0.11 +/- 0.04% of the amplitude of the peak current. 8. Late openings of the Na+ channels were frequent in patches on the myotonic fibre segments. The amplitude of the current produced by re-openings was as high as about 0.75 +/- 0.11% of the amplitude of the peak current.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Characteristics of Na+ channels and Cl- conductance in resealed muscle fibre segments from patients with myotonic dystrophy. 169 78

To determine the response of cardiac Na current (INa) in adult cardiac ventricular myocytes to culture, single isolated ventricular myocytes from collagenase-perfused adult cat hearts were placed in primary culture for up to 2 wk on a two-dimensional (2D) surface (laminin-coated coverslips), which allowed the morphology of the myocytes to change markedly, or in a three-dimensional matrix (3D) of alginate, in which cell shape changed only minimally. Action potentials and INa were recorded from groups of 1) freshly isolated myocytes serving as the control (day 0),2) cells maintained in 2D culture for 9-14 days (2D, day 9-14), and 3) cells cultured in alginate for 9-14 days (3D, day 9-14) with use of a conventional whole cell patch technique. Maximal upstroke velocity (Vmax) of the action potential was reduced by approximately 50% in 2D- and 3D-cultured cells relative to controls. INa in 2D- and 3D-cultured cells was strikingly different from that in control myocytes. Half-maximal voltage (V 1/2) for the chord conductance-voltage relationship was shifted approximately 15 mV negatively to that for controls in 2D- and 3D-cultured cells. INa steady-state availability curve also shifted negatively relative to controls in 2D- and 3D-cultured myocytes, but the magnitude of this shift (approximately 16-20 mV) was greater than that for the chord conductance-voltage curve.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Alteration of the sodium current in cat cardiac ventricular myocytes during primary culture. 773 48

We measured macroscopic sodium currents (INa) in preparations from adult rat ventricle under four different conditions (I-IV): using the cell attached configuration of the tight-seal patch clamp technique on cells isolated with either trypsin followed by collagenase (I) or with collagenase only (II), and using the loose patch technique on cells isolated with collagenase (II) as well as on multicellular preparations not subjected to enzyme treatment (IV). The voltage dependence of the steady-state activation of INa as well as of the steady-state inactivation differed significantly among condition I and II. Moreover, the recordings were voltage shifted in comparison to the recording condition III and IV. The potentials of half maximal activation and inactivation were: [sequence data: see text] The shift of inactivation was time dependent and continued after 3-5 min after the seal formation in condition I, but not in condition II. No time dependent shift was found in III and IV. We conclude, that the voltage dependence of cardiac sodium current is shifted by gigaseal patch recording. The degree of this shift depends on the type of enzymatic isolation procedure, with trypsin causing more pronounced effects than collagenase. The cell isolation itself seems not to interfere with the voltage dependence of INa, since loose patch recordings from multicellular preparations and from single cells isolated with collagenase show no obvious differences.
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PMID:Influence of cell isolation and recording technique on the voltage dependence of the fast cardiac sodium current of the rat. 779 48

The effects of metabolic inhibition on K+ background currents and action potential duration were investigated in neonatal rat ventricle cells during early development. Action potentials and ionic currents were measured with the patch clamp technique in current and voltage clamp mode in cells isolated with collagenase from 1 day and 7 day old rats. During the first postnatal week, the cell surface increased from 1700 to 2210 microm2 and the membrane hyperpolarized from -66.1 to -72.0 mV. Concomitantly the action potential shortened and the plateau became more negative. Inhibition of oxidative phosphorylation (50 microM 2,4 DNP) or of glycolysis in 1 day old rats (5 mM 2-deoxyglucose, 2-DG) also shortened the action potential by about 50% after 5 min exposure. The background current measured in the absence of INa, ICa,L, and Ito included: (1) an inward rectifying component whose I/V curves crossed over when measured in 6, 15, or 30 mM [K]o and showed an increase in slope conductance when [K]o was raised. Inward rectification was abolished by 2.4 mM Ba2+ in 1 day old cells and by 0.2 mM one week after birth; (2) a glibenclamide (100 microM) sensitive component that developed with time after membrane rupture (5-10 min) showing a higher current density in 7 than in 1 day old animals (1.4 vs 0.2 microA x cm-2 at -50 mV); and (3) a small and almost linear leak component of comparable amplitude in both age groups. Inhibition of oxidative phosphorylation with 2.5 microM carbonylcyanide m-chlorophenylhydrazone induced the development of background currents with different properties in both age groups: An inwardly rectifying Ba2+ sensitive current in 1 day old cells and a glibenclamide sensitive outwardly rectifying current in the 7 day old group. In contrast, exposure to 5 mM 2-DG provoked in all cells the development of an outwardly rectifying current that was blocked by glibenclamide. We conclude that the electrophysiologic response to metabolic inhibition is determined by the relative importance of the metabolic pathways present which in turn depends on the developmental state of the cells.
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PMID:Background K+ currents and response to metabolic inhibition during early development in rat cardiocytes. 945 Jun 58