Locust Skeletal Muscle Research Articles

Slow motor neuron stimulation of locust skeletal muscle: model and measurement

The isometric force response of the locust hind leg extensor tibia muscle to stimulation of a slow extensor tibia motor neuron is experimentally investigated, and a mathematical model describing the response presented. The measured force response was modelled by considering the ability of an existing model, developed to describe the response to the stimulation of a fast extensor tibia motor neuron and to also model the response to slow motor neuron stimulation. It is found that despite large differences in the force response to slow and fast motor neuron stimulation, which could be accounted for by the differing physiology of the fibres they innervate, the model is able to describe the response to both fast and slow motor neuron stimulation. Thus, the presented model provides a potentially generally applicable, robust, simple model to describe the isometric force response of a range of muscles.

A comparison of models of the isometric force of locust skeletal muscle in response to pulse train inputs

Muscle models are an important tool in the development of new rehabilitation and diagnostic techniques. Many models have been proposed in the past, but little work has been done on comparing the performance of models. In this paper, seven models that describe the isometric force response to pulse train inputs are investigated. Five of the models are from the literature while two new models are also presented. Models are compared in terms of their ability to fit to isometric force data, using Akaike's and Bayesian information criteria and by examining the ability of each model to describe the underlying behaviour in response to individual pulses. Experimental data were collected by stimulating the locust extensor tibia muscle and measuring the force generated at the tibia. Parameters in each model were estimated by minimising the error between the modelled and actual force response for a set of training data. A separate set of test data, which included physiological kick-type data, was used to assess the models. It was found that a linear model performed the worst whereas a new model was found to perform the best. The parameter sensitivity of this new model was investigated using a one-at-a-time approach, and it found that the force response is not particularly sensitive to changes in any parameter.

Activities of octopamine and synephrine stereoisomers on octopaminergic receptor subtypes in locust skeletal muscle.

The activities of the (-) and (+)- forms of p-, m- and o-octopamine and p- and m-synephrine have been compared on the different subtypes of octopamine receptor present in the extensor-tibiae neuromuscular preparation from the locust hindleg. The rank order of potency of the (-)-forms on the OCTOPAMINE2A receptors was p-synephrine greater than p-octopamine greater than m-octopamine greater than o-octopamine greater than m-synephrine whilst the rank order of the (+)-forms was p-synephrine greater than p-octopamine greater than m-octopamine. (+)-m-Synephrine and (+)-o-octopamine had no effect on this class of receptor when tested up to a concentration of 10(-3) M. The rank order of potency of the (-)-forms on the OCTOPAMINE2B receptors was p-synephrine greater than p-octopamine greater than m-synephrine greater than m-octopamine greater than o-octopamine whilst the rank order of the (+)-forms was p-octopamine greater than p-synephrine greater than m-octopamine greater than o-octopamine. (+)-m-Synephrine again had no effect up to a concentration of 10(-3) M. The rank order of potency of the (-)-forms on the OCTOPAMINE1 receptors was p-synephrine greater than p-octopamine greater than m-synephrine greater than m-octopamine greater than o-octopamine, whilst the rank order of the (+)-forms was p-synephrine greater than p-octopamine greater than o-octopamine greater than m-synephrine greater than m-octopamine.(ABSTRACT TRUNCATED AT 250 WORDS)

Modelling the isometric force response to multiple pulse stimuli in locust skeletal muscle

An improved model of locust skeletal muscle will inform on the general behaviour of invertebrate and mammalian muscle with the eventual aim of improving biomedical models of human muscles, embracing prosthetic construction and muscle therapy. In this article, the isometric response of the locust hind leg extensor muscle to input pulse trains is investigated. Experimental data was collected by stimulating the muscle directly and measuring the force at the tibia. The responses to constant frequency stimulus trains of various frequencies and number of pulses were decomposed into the response to each individual stimulus. Each individual pulse response was then fitted to a model, it being assumed that the response to each pulse could be approximated as an impulse response and was linear, no assumption were made about the model order. When the interpulse frequency (IPF) was low and the number of pulses in the train small, a second-order model provided a good fit to each pulse. For moderate IPF or for long pulse trains a linear third-order model provided a better fit to the response to each pulse. The fit using a second-order model deteriorated with increasing IPF. When the input comprised higher IPFs with a large number of pulses the assumptions that the response was linear could not be confirmed. A generalised model is also presented. This model is second-order, and contains two nonlinear terms. The model is able to capture the force response to a range of inputs. This includes cases where the input comprised of higher frequency pulse trains and the assumption of quasi-linear behaviour could not be confirmed.

Isometric force generated by locust skeletal muscle: responses to single stimuli

A mathematical model of the locust hind leg extensor muscle is described. The model accounts for the force response of the muscle to well-separated input stimuli under isometric conditions. Experimental data was collected by stimulating the extensor muscle and measuring the force generated at the tibia. In developing a model it was assumed that the response to a single isolated stimulus was linear. A linear model was found to fit well to the response to an isolated stimulus. No assumptions were made about the model order and models of various order were fitted to data in the frequency domain, using a least squares fit. The stimulus can be approximated as an impulse, with the response to each stimulus well described by a linear second-order system. Using a third-order model provided a better fit to data, but the improvement in fit was marginal and the model uses one extra parameter. A fourth-order model, which is often used to describe the behaviour of isometric muscle was found to overfit the data. Using a second-order model provides a simpler way of describing the behaviour of an isometric twitch.

Ryanoids change the permeability of potassium channels of locust (Schistocerca gregaria) muscle.

The plasma membrane of locust skeletal muscle contains two types of K+ channel; a maxi, Ca2+-activated 170 pS channel (BK channel) and an inward rectifier of 35 pS conductance (IK channel). The effects of ryanodine, 9,21-didehydroryanodine and 9,21-didehydroryanodol on these channels have been investigated. In the concentration domain 10(-9) M to 10(-5) M, ryanodine irreversibly induced a dose-dependent reduction of the reversal potential (Vrev) of the channel currents, measured under physiologically normal K+ and Na+ gradients, i.e. from approximately 60 mV in the absence of ryanodine to approximately 15 mV for 10(-5) M ryanodine. The alteration of K+ channel selectivity was Ca2+ independent. 9,21-Didehydroryanodine and 9,21-didehydroryanodol reduced Vrev, but only to approximately 35 mV during application of 10(-5) M of these compounds. 9,21-Didehydroryanodine also diminished the conductances of the locust K+ channels. The three ryanoids reduced Vrev of a Ca2+-activated, high-conductance channel in inside-out patches excised from mouse interosseal muscle, although the changes were in each case less pronounced than those for the locust K+ channels. Also, the action of 9,21-didehydroryanodine on mouse K+ (BK) channels was restricted to a shift of Vrev. For all three ryanoids, the IC50 coefficients (i.e. the concentration of toxin that gave a 50% reduction in Vrev) for the shifts in Vrev were similar for the locust and mouse muscle K+ channels.

Novel actions of ryanodine and analogues--perturbers of potassium channels.

The effects of ryanodine, 9,21-didehydroryanodine and 9,21-didehydroryanodol on two types of K(+) channel (a maxi, Ca(2+)-activated, 170pS channel (BK channel) and an inward rectifier, stretch sensitive channel of 35 pS conductance (IK channel)found in the plasma membrane of locust skeletal muscle have been investigated. 10(-9) M-10(-5) M ryanodine irreversibly induced a dose-dependent reduction of the reversal potential (V (rev)) of the currents of both channels, i.e. from 60 mV in the absence of the alkaloid to 15 mV for 10(-5) M ryanodine, measured under physiologically normal K(+) and Na(+) gradients. In both cases the change in the ionic selectivity was Ca(2+) -independent. 9,21-didehydroryanodine and 9,21-didehyroryanodol also reduced V (rev), but only to 35 mV during application of 10(-5) M of these compounds. Additionally, 9,21-didehydroryanodine reversibly diminished the conductances of the two K(+) channels. To test the hypothesis that ryanoids increase Na permeability by enlarging the K(+) channels, the channels were probed with quaternary ammonium ions during ryanoid application. When applied to the cytoplasmic face of inside-out patches excised from locust muscle membrane, TEA blocked the K(+) channels in a voltage-dependent fashion. The dissociation constant (K (d)(0)) for TEA block of the IK channel was reduced from 44 mM to 1 mM by 10(-7) M ryanodine, but the voltage-dependence of the block was unaffected. Qualitatively similar data were obtained for the BK channel. Ryanodine had no effect on the K (d) for cytoplasmically-applied TMA. However, the voltage-dependence for TMA block was increased for both K(+) channels, from 0.47 to 0.8 with 10(-6) M ryanodine. The effects of ryanodine on TEA and TMA block support the hypothesis that ryanodine enlarges the K(+) channels so as to facilitate permeation of partially hydrated Na(+) ions.

The action of philanthotoxin-343 and photolabile analogues on locust (Schistocerca gregaria) muscle.

The effects of philanthotoxin-343 (PhTX-343; tyrosyl-butanoyl-spermine) and photolabile analogues of this synthetic toxin on locust (Schistocerca gregaria) skeletal muscle have been investigated using whole muscle preparations (twitch contractions), single muscle fibres (excitatory postsynaptic currents (EPSCs)) and muscle membrane patches containing single quisqualate-sensitive glutamate receptors (qGluR). Analogues containing an azido group attached to either the butanoyl side-chain of PhTX-343 or as a substitute for the hydroxyl moiety of the tyrosyl residue were about 6 fold more potent antagonists than PhTX-343; those with an azido group located at the distal end of the toxin molecule were generally 2-3 fold less potent than PhTX-343. When these compounds were tested in subdued light, they were reversible antagonists of the muscle twitch, EPSC and qGluR. When a muscle was irradiated with U.V. during application of photolabile toxin combined with either neural stimulation of the muscle or L-glutamate application, antagonism of the twitch, EPSC and qGluR was complete and irreversible.

Identification of proteins resembling G-protein alpha subunits in locust muscle

In locust skeletal muscle, FMRFamide-like peptides decrease a K+ conductance. Functional data suggest the involvement of G-proteins. For identification of G-protein alpha-subunits, membranes of locust skeletal muscle were probed with ADP-ribosylating bacterial toxins, the photoreactive GTP analog, [alpha-32P]GTP azidoanilide, and with antibodies against mammalian alpha-subunits. Multiple guanine nucleotide-binding proteins of approximately 24-95 kDa were detected. Pertussis toxin catalyzed the ADP-ribosylation of two proteins comigrating with the ADP-ribosylated alpha-subunits of the mammalian G-proteins Go and Gi. Cholera toxin promoted ADP-ribosylation of a protein comigrating with mammalian cholera toxin substrates (i.e., Gs alpha-subunits). An antibody against mammalian Go alpha-subunits detected a 54-kDa protein. Thus proteins with properties of mammalian G-protein subunits are present in insect muscle.

Hyperpolarization slowly activates a potassium current in locust skeletal muscle

An inward current activated by hyperpolarization, IK,H, was studied under voltage clamp in locust skeletal muscle. The dependence of its reversal potential on [K+]o and its insensitivity to changes in [Na+]o indicate that the underlying conductance is a K+ conductance. The instantaneous current-voltage (I-V)-relationship exhibits outward rectification. Activation and deactivation take seconds and have complex time courses. At 10 mM [K+]o activation seems to start at a voltage greater than or equal to 20 mV more positive than the resting potential (approximately -68 mV). Ba++ blocks IK,H strongly; so do Rb+ and Cs+, the latter in a voltage dependent manner. The slow inward current bears similarities to anomalous rectifiers as well as to mixed, hyperpolarization-activated K+/Na+ currents in other tissues.

A concentration-dependent localization of octopamine-sensitive adenylate cyclase activity in locust skeletal muscle

A concentration-dependent localization of octopamine-sensitive adenylate cyclase activity has been demonstrated in skeletal muscle of the locust, Schistocerca gregaria, using an histochemical technique. In the intermediate speed contracting muscle fibres from the fan region of the extensor-tibiae muscle of the locust hindleg, low concentrations of DL-octopamine (10(-8) M) induce reaction product preferentially in the sarcoplasmic reticular component of the dyads. At slightly higher concentrations (10(-7) and 10(-6) M) lower amounts of diffuse reaction product are also found in the non-dyad sarcoplasmic reticulum and at the sarcolemmal membrane, with occasional amounts of a less diffuse, punctuate product in the transverse tubule (T-tubule) component of the dyads. At higher concentrations (10(-5) and 10(-3) M) the predominant product is the dense, plaque-like accumulations of reaction product in the T-tubule component of the dyads. The results are discussed in terms of the likely physiological significance of the accumulation of reaction product in these different locations.

Ion-Selectivity of Single Glutamate-Gated Channels in Locust Skeletal Muscle

ABSTRACT The ion-selectivity of the extrajunctional glutamate-gated ion channel in locust extensor tibiae muscle was studied using the patch-clamp technique. The alkali metal ions Li+, Na+, K+, Rb+ and Cs+ were all highly permeant, with reversal potentials close to 0mV. Both complete and partial replacement of Na+ (180 mmol I−1 in standard saline) showed that conductance (γ) increased in the order Li+< Na+< Cs+< Rb+ (approx. 70–125 pS), γK being close to γCs. The channel was impermeable to the large organic monovalent ions tetramethyl ammonium, guanidinium and choline, and permeable to the smaller ammonium ion. Divalent cations (Ca2+ and Mg2+) did not contribute measurably to the ionic current. Indications were obtained that high concentrations of Mg2+ or Ca2+ block the channel. The results suggest that the glutamate-gated channel combines a high conductance with a restricted ion-selectivity, based on ion charge and size, the conductance being dependent on the dehydration energy of the ionic species.

Histochemical localization of octopamine- and proctolin-sensitive adenylate cyclase activity in a locust skeletal muscle.

A histochemical technique for the localization of adenylate cyclase activity has been applied to the extensor-tibiae muscle of the hindleg of the locust, Schistocerca gregaria to localise the sites of action of the modulatory compounds octopamine and proctolin. Octopamine-sensitive adenylate cyclase activity can be demonstrated in fast and intermediate type muscle fibres but not in the limited number of purely slow muscle fibres (3-6) in the fan region at the proximal end of the muscle. In contrast the latter fibres are the only ones in the muscle to exhibit proctolin-sensitive adenylate cyclase activity. In both cases the bulk of the reaction product is localised in the sarcoplasmic reticulum component of the dyads, with lesser amounts occurring beneath the sarcolemmal membrane, in the non-dyad sarcoplasmic reticulum and in the T-tubule system. The results are consistent with physiological data suggesting that proctolin, but not octopamine, mediates its effects on the myogenic rhythm of contraction and relaxation in this muscle by changing the levels of cyclic AMP in the small group of slow muscle fibres which act as the pacemaker for this rhythm.

Resting Potential and Potassium-Selective Electrode Measurements in Locust Skeletal Muscles

ABSTRACT The membrane of locust skeletal muscle fibres is permeable to both potassium and chloride ions in the resting state and has a potassium selectivity similar to that of a potassium electrode (Usherwood, 1967). The experiments described here were performed on the isolated retractor unguis muscle of the locust Schistocerca gregaria. The composition of the standard locust saline was similar to that of Hoyle (1953) except that in most experiments the HPO4/HCO3 buffer was replaced by Tris-maleate (pH 6·8). Conventional electrophysiological techniques were used and electrodes had tip potentials of less than 5 mV.

Regional differences in responsiveness to octopamine within a locust skeletal muscle.

Regional differences in the physiological and biochemical responses to octopamine have been investigated in the extensor tibiae muscle of the hind leg of the locust. Octopamine increases the rate of relaxation of twitch tension generated by the slow motoneurone by different amounts in different regions of the muscle. It also increases the amplitude of twitch tension by the same amount in different regions of the muscle. The relaxation rate of fast twitch tension is increased by the same amount in all regions of the muscle innervated by the fast motoneurone. Octopamine also increases cyclic AMP levels by different amounts in different regions of both the extensor muscle and its antagonistic muscle, the flexor tibiae. In both muscles the maximal responses are obtained in the regions of the muscles containing the highest proportions of slow and intermediate muscle fibres as characterized by their contractile and ultrastructural properties. The results are discussed in terms of their functional significance and compared with examples of differential responses of different muscle fibre types in other invertebrate and vertebrate skeletal muscles.

Inhibition of GABA uptake potentiates the effects of exogenous GABA on locust skeletal muscle

1. Insect skeletal muscle is relatively insensitive to applied GABA, responses are elicited only when relatively high concentrations of GABA are used (>10 −6M). 2. Pretreatment of the muscle with the GABA uptake inhibitors nipecotic acid, β-aminobutyric acid or β-alanine increases the sensitivity of the muscle to GABA by as much as 1000-fold. 3. The evidence suggests the existence of a GABA uptake mechanism in the insect neuromuscular system which could reside in glial cells.

Synaptic and non-synaptic effects of molluscan cardioexcitatory neuropeptides on locust skeletal muscle

Neurally evoked contractions of the locust jumping muscle are potentiated by the molluscan cardioexcitatory peptide (FMRFamide) and related peptides. This effect is due to a synaptic and a non-synaptic action of the peptides: (1) excitatory synaptic potentials are increased; (2) excitation-contraction coupling becomes more efficient, as indicated by an increased strength of potassium contractures. Octopus heptapeptide (YGGFMRFamide), which contains the entire enkephalin structure (YGGFM), is 100 times more potent than FMRFamide. Structure-activity studies indicate that the observed actions are not mediated by opioid receptors but are dependent on the C-terminal structure of the peptides.

The role of cyclic nucleotides and calcium in the mediation of the modulatory effects of octopamine on locust skeletal muscle.

The role of cyclic AMP and calcium in the mediation of the effects of octopamine has been investigated in the extensor tibiae muscle of the hind leg of the locust. Elevation of cyclic AMP levels in the preparation by means of the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX), by means of the diterpene adenylate cyclase activator, forskolin, and by means of cyclic nucleotide analogues mimics the post-synaptic effects of octopamine application at different frequencies of neuronal stimulation. These conditions also mimic the presynaptic effects of octopamine on spontaneous release of transmitter from the slow motoneurone. The effects of octopamine on the preparation are calcium sensitive, with the maximal sensitivity occurring between 0.5 and 4.0 mM-external calcium. The results are discussed in terms of the role of cyclic AMP and calcium in the mediation of the effects of octopamine.

A modulatory octopaminergic neurone increases cyclic nucleotide levels in locust skeletal muscle.

Octopamine increases the level of cyclic AMP in a dose-dependent way in the locust extensor tibiae neuromuscular preparation. The response peaks after a 10 min exposure and then declines to a plateau. The effect of octopamine is potentiated in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). The levels of cyclic GMP in the muscle were not affected by octopamine. The response is stereospecific for the naturally occurring D(-) isomer of octopamine and is also specific for monophenolic biogenic amines. Studies with a range of synthetic agonists and antagonists reveal that the receptors mediating the response are of the OCTOPAMINE2 class. Forskolin, a diterpene activator of adenylate cyclase activity, increases cyclic AMP but not cyclic GMP levels in the extensor muscle. The response has a prolonged time course and is again potentiated by IBMX. Stimulation of the octopaminergic neurone to the extensor muscle increases the levels of cyclic AMP but not those of cyclic GMP. The response is blocked by phentolamine, an alpha-adrenergic blocking agent that also blocks the effects of octopamine in this preparation. The results are discussed in terms of the parallels between the biochemical and physiological effects of octopamine on this muscle and in terms of the mode of action of the octopamine receptors present.

Block of synaptic vesicle exocytosis without block of ca 2+-influx. An ultrastructural analysis of the paralysing action of habrobracon venom on locust motor nerve terminals

The venom of the wasp Habrobracon hebetor presynaptically blocks excitatory but not inhibitory neuromuscular transmission at locust skeletal muscle. Its mode of action on excitatory motor nerve terminals has been studied at the retractor unguis muscle of Schistocerca by means of ultrastructural stereology. Paralysed and unparalysed preparations, either resting or stimulated for 7 min at 20 Hz, were compared. Paralysis does not cause structural damage to the nerve terminals but prevents the depletion of vesicles occurring upon nerve stimulation in the controls. Prolonged paralysis leads to an increase in the number and the size of vesicles resulting in an increase of total membrane per terminal cross-section by about 33% after 2 days. Stimulation causes swelling of mitochondria both in controls and in paralysed preparations, resulting from a rise of intraterminal [Ca 2+] as is indicated by the absence of the swelling if extracellular Ca 2+ is replaced by Mg 2+. In addition, stimulation leads to a reduction of vesicle size, an increase in the area of axolemma and in the number of cisternae and of profiles of the smooth endoplasmic reticulum in controls but not in paralysed preparations. However, neither in controls nor in paralysed preparations is the total amount of membrane per teminal cross-section affected by stimulation. Under paralysis, vesicles tend to stick to the presynaptic membrane. It is concluded that Habrobracon venom does not block the depolarization-dependent Ca 2+-influx into the nerve terminal and that it is unlikely to interfere with some transmitter-related process. Rather, the venom seems to block vesicle exocytosis itself. The results lend further support to the view that in insect neuromuscular synapses exocytosis is the mechanism whereby transmitter quanta are released.