Octopaminergic innervation and modulation of a locust flight steering muscle

We demonstrate that the meso- and metathoracic pleuroaxillary flight steering muscle (M85 mesothorax, M114 metathorax) of the migratory locust are each innervated by a single dorsal unpaired median neurone (DUM3,4,5a). The soma of this neurone can be localized by retrograde staining of the motor nerve with Neurobiotin, but not with cobalt salts. The primary neurite projects in the superficial DUM cell tract, and the axons run in nerve roots 3, 4 and 5 and in all their secondary branches. Other muscle targets include the second tergal remotor coxa (M120) and the posterior rotator coxae (M122, M123, M124), but not the first tergal remotor coxa (M119) and subalar (M129) flight muscles. Octopamine-like immunoreactive varicosities occur on the pleuroaxillary muscles. Stimulation of DUM3,4,5a and octopamine (10(-6) mol l-1) superfusion increased the amplitude and the relaxation velocity of neurally evoked twitch contractions of this muscle. Octopamine also significantly reduced the tonic tension that this muscle develops when stimulated at flight frequency (20 Hz), while increasing the amplitude of each phasic twitch. A catch-like tension is also reduced in the presence of octopamine. Simulations of the motor pattern experienced by the pleuroaxillary muscles during roll manoeuvres suggest that transient changes in tension underlying corrective steering could be doubled in the presence of octopamine.

Effects of atriopeptin III on isolated mesenteric resistance vessels from SHR and WKY

The effects of atriopeptin III (AP III) were determined on agonist-induced [i.e., 10(-4) M norepinephrine (NE)] and depolarization-induced (80 mM K+) contractions of isolated mesenteric resistance vessels (ID approximately 100 microns) from spontaneously hypertensive rats (SHR) and from normotensive control Wistar-Kyoto (WKY) rats. The vessels from both groups, when activated by 80 mM K+, were unaffected by AP III. However, activation of WKY vessels by 10(-4) M NE (both phasic and tonic contraction) was inhibited quite effectively and potently by AP III, whereas that in SHR vessels was much less inhibited. In the WKY rat vessels, the concentration of AP III that inhibited contraction by 50% for NE-induced phasic tension was 3.1 +/- 1.3 nM, whereas in SHR vessels it was nearly 1 microM. Comparison of AP III inhibition of NE-induced phasic tension to that at 5 min of activation (tonic tension) indicated that the tonic contractions were less sensitive to AP III than the phasic contractions in the vessels from both strains. A similar experiment indicated that AP III was a potent inhibitor of agonist-induced activation in a human renal resistance vessel (ID 125 microns) and that this vessel depended virtually completely on extracellular Ca2+ for NE-induced contraction. These studies contrast with earlier reports (1, 30) that similar peptides inhibited tension only in rat renal resistance vessels and not in resistance vessels from other vascular beds. The decreased sensitivity and efficacy of AP III in inhibiting tension in SHR compared with WKY mesenteric resistance vessels is discussed in the context of the etiology of spontaneous hypertension.

Dependence of tonic tension on extracellular calcium in rat extraocular muscle

Ca2+-free saline containing 3.0 mM Mg2+ virtually abolishes the tonic tension evoked by depolarization with a high K+ concentration of the tonic multiply innervated fibers of rat extraocular muscles. The tonic tension abolished by Ca2+ withdrawal is restored when Ca2+ is substituted by Sr2+ but not by Ni2+. The increase of Mg2+ reduces the tonic tension and displaces the tension-log K+ relationship to the right. Cd2+ significantly reduces the tension amplitude but does not shift the tension-log K+ relationship. The organic blocker of Ca2+ channels, nifedipine (1-10 microM), has no effect on the tonic tension. In contrast, diltiazem (20 microM) reduces the amplitude of the responses without changing the tension-log K+ relationship. Both foreign anions NO3- and SCN- potentiate tonic tension without changing the tension-log K+ relationship. SCN- increases the resting tension of the muscle; this effect depends on Ca2+. In conclusion, the disappearance of tonic tension after Ca2+ withdrawal is not due to depolarization of the fibers or inactivation of the contractile responses. It is suggested that entry of extracellular Ca2+, via a voltage-dependent Ca2+ conductance, or specific interactions of Ca2+ with membrane sites involved in the regulation of excitation-contraction coupling play a role in evoking tension in tonic fibers.

Effects of changes of intracellular pH on contraction in sheep cardiac Purkinje fibers.

Intracellular pH (pHi) was measured with a pH-sensitive microelectrode in voltage-clamped sheep cardiac Purkinje fibers while tension was simultaneously measured. All solutions were nominally CO2/HCO3 free and were buffered with Tris. The addition of NH4Cl (5-20 mM) produced an initial intracellular alkalosis that was associated with an increase of twitch tension. At the same time, a component of voltage-dependent tonic tension developed. Prolonged exposure (greater than 5 min) to NH4Cl resulted in a slow recovery of pHi accompanied by a decrease of tension. Removal of NH4Cl produced a transient acidosis that was accompanied by a fall of force. In some experiments, there was then a transient recovery of force. If extracellular pH (pHo) was decreased, then pHi decreased slowly. Tension also fell slowly. An increase of pHo produced a corresponding increase of both force and pHi. The application of strophanthidin (10 microM) increased force and produced an intracellular acidosis. The addition of NH4Cl, to remove this acidosis partially, produced a significant increase of force. The above results show that contraction is sensitive to changes of intracellular but not extracellular pH. This pH dependence will therefore modify the contractile response to inotropic maneuvers that also affect pHi.

Role of the different calcium sources in the excitation–contraction coupling in crab muscle fibers

Excitation–contraction coupling in crab muscle fibers was studied in voltage-clamp conditions. Extracellular calcium is essential for the mechanical activity. Two calcium influxes induced by membrane depolarization contribute to tension development: one is the inward calcium current responsible for the phasic tension, the other is a calcium influx dependent on extracellular sodium and calcium concentrations and is responsible for the tonic tension. These calcium influxes are not sufficient to activate contractile proteins. Experiments with procaine and caffeine show that a calcium release from the sarcoplasmic reticulum is required.

The effect of D600 on tonic tension, Na+ inward current, and Na+–Ca2+ exchange in frog heart

Preparations of frog atrial muscle were stimulated at 0.33 Hz under voltage clamp, and the resulting membrane currents and the twitch contractions (phasic and tonic components) were recorded in presence or absence of D600. It has been suggested earlier that the tonic contractions are regulated by an electrogenic Na+–Ca2+ exchange, while the phasic contractions are closely related to the calcium inward current (Isi). In this study we investigated the effect of D600 on (i) the tonic contractions elicited by long depolarizing pulses of high amplitude and (ii) the tonic contractions increased by veratrine and resulting in a positive inotropic effect (PIE). While 1 μM D600 reduced Isi and the corresponding phasic contractions to < 30% of their initial values within 5 min, the inhibitory effect of D600 on tonic contractions developed more slowly or higher concentrations of D600 were needed to achieve similar levels of inhibition within the same time. Furthermore, applications of 5–50 μM D600 inhibited the veratrine-induced increase in INa and in tonic contractions, and both of these effects again fully developed within a few minutes of D600 being removed. The results demonstrate that D600 inhibits not only Isi and phasic contractions, but it also decreases the tonic contractions in frog heart. The effect on the tonic component is associated with inhibition of the tetrodotoxin-sensitive Na+ inward current, and the results are interpreted as an effect of D600 on the electrogenic Na+–Ca2+ exchange. These additional effects of D600 should be considered when using this drug as the "specific" calcium channel blocker.