Mechanisms controlling venom expulsion in the western diamondback rattlesnake, Crotalus atrox

Although many studies have documented variation in the amount of venom expended during bites of venomous snakes, the mechanistic source of this variation remains uncertain. This study used experimental techniques to examine how two different features of the venom delivery system, the muscle surrounding the venom gland (the Compressor Glandulae in the rattlesnake) and the fang sheath, could influence venom flow in the western diamondback rattlesnake, Crotalus atrox. Differential contraction of the Compressor Glandulae explained only approximately 30% of the variation in venom flow. Lifting (compression) of the fang sheath as occurs during a normal strike produced marked increases in venom flow; these changes were closely correlated and exceed in magnitude by almost 10 x those recorded from the Compressor Glandulae alone. These results suggest that variation in these two aspects of the venom delivery system--both in terms of magnitude and temporal patterning--explain most of the observed variation in venom injection. The lack of functional or mechanical links between the Compressor Glandulae and the fang sheath, and the lack of skeletal or smooth muscle within the fang sheath, make it unlikely that variation in venom flow is under direct neural control. Instead, differential venom injection results from differences in the pressurization by the Compressor Glandulae, the gate keeping effects of the fang sheath and enclosed soft-tissue chambers, and by differences in the pressure returned by peripheral resistance of the target tissue.     

Funnel web spider venom produces spontaneous action potentials in nerve

Venom from the lethal Australian spider, $\text{Atrax robustus}$, causes fasciculation of muscles $\text{in vivo}$ and in isolated diaphragms in mice. Spontaneous end-plate potentials were recorded in muscle fibres exposed to the venom and associated spontaneous electrical activity could also be recorded from the phrenic nerve. It was proposed that the venom produces muscle fasciculation by causing abnormal, spontaneous, repetitive firing of motor nerves. The mechanism of this action was investigated in aplysia neurones. The venom produced abnormal, spontaneous, repetitive inward currents in voltage clamped neurones and changed the current-voltage characteristics of the surface membrane. It is suggested that the basic mode of action of Funnel-web venom is to change the electrical field in nerve membrane.     

Exercise and glycogen depletion: effects on ability to activate muscle phosphorylase

Phosphorylase activation reverses during prolonged contractile activity. Our first experiment was designed to determine whether this loss of ability to activate phosphorylase by stimulation of muscle contraction persists following exercise. Phosphorylase activation by stimulation of muscle contraction was markedly inhibited in rats 25 min after exhausting exercise. To evaluate the role of glycogen depletion, we accelerated glycogen utilization by nicotinic acid administration. A large difference in muscle glycogen depletion during exercise of the same duration did not influence the blunting of phosphorylase activation. Phosphorylase activation by stimulation of contraction was more severely inhibited following prolonged exercise than after a shorter bout of exercise under conditions that resulted in the same degree of glycogen depletion. A large difference in muscle glycogen repletion during 90 min of recovery was not associated with a significant difference in the ability of muscle stimulation to activate phosphorylase, which was still significantly blunted. Phosphorylase activation by epinephrine was also markedly inhibited in muscle 25 min after strenuous exercise but had recovered completely in glycogen-repleted muscle 90 min after exercise. These results provide evidence that an effect of exercise other than glycogen depletion is involved in causing the inhibition of phosphorylase activation; however, they do not rule out the possibility that glycogen depletion also plays a role in this process.     

Early effect of Crotalus durissus terrificus venom on kidney circulation

Local blood flow was measured in renal cortex (1 mm below cortical surface) by means of the hydrogen clearance method in urethanized rats. Recording of blood pressure from femoral artery was performed. Crotalus durissus terrificus venom injection (one mg/kg i.v.) significantly decreased cortical blood flow at 10 min, without a significant arterial pressure modification. Posterior injection of mannitol 200 mg induced a significant increase in cortical blood flow, although initial values were not reached. Electron microscopy showed thromboses in the glomerular capillaries 35 minutes after venom injection. It is suggested that the precocious effect of this venom on renal cortical blood flow may be instrumental in the development of the renal acute insufficiency induced by Crotalus durissus terrificus venom.     

Exogenous testosterone treatment decreases diaphragm neuromuscular transmission failure in male rats.

The effect of chronic exogenous testosterone (T) treatment on neuromuscular transmission in the diaphragm (Dia) muscle of adult male rats was determined. The contribution of neuromuscular transmission failure (NTF) to Dia fatigue was evaluated by superimposing intermittent direct muscle stimulation on repetitive nerve stimulation of isometric contraction in vitro. T treatment significantly reduced the contribution of NTF to Dia fatigue by approximately 20% (P < 0.001). Fiber type-specific effects on NTF were determined by measuring Dia fiber glycogen levels subsequent to repetitive nerve or muscle stimulation. T treatment had no effect on glycogen depletion in Dia type I and IIa fibers regardless of stimulation route. In the control group, type IIx fibers demonstrated significantly less glycogen depletion after nerve stimulation compared with direct muscle stimulation (P < 0.05), suggesting the presence of NTF. In contrast, T treatment increased glycogen depletion of type IIx fibers during nerve stimulation to levels similar to those after direct muscle stimulation. These data indicate that testosterone treatment substantially improves neuromuscular transmission in the Dia.     

Glycogen degradation during isometric exercise at low contraction force

The glycogen content was measured in biopsy sample of human vastus lateralis muscle during prolonged isometric contraction with low force generation. In the first experiment 15% of the maximum voluntary contraction force (MVC) was held for 10 min. Glycogen utilization was 68.1 mmol glucosyl units.kg-1 dry muscle (d.m.). The study was continued by intermittent contractions of 50 s duration and 10 s rest repeated for 50 min. This resulted in a total glycogen utilization of 167.5 mmol glycosyl units.kg-1 d.m. The study was repeated with a force set of 7.5% MVC starting with 20 min continuous contraction followed by the same intermittent contractions for a further 100 min. The glycogen decrease was 15 mmol after the continuous contraction and totally 50 mmol after 2 h with the lower force. Thus the glycogen degradation rate even at low contraction force was related to the force level, being 6 times higher when the force was increased from 7.5 to 15% MVC. With prolonged isometric work periods at work loads corresponding to 15% MVC or higher depletion of the glycogen store can limit work performance capacity.     

In vivo actions of atraxin, a protein neurotoxin from the venom glands of the funnel-web spider (Atrax robustus)

The effects of atraxin, a neurotoxic protein from the venom glands of the funnel-web spider (Atrax robustus), have been studied in anaesthetized monkeys. At doses of 70 and 80 micrograms kg-1 i.v., atraxin caused respiratory disturbances (dyspnoea and apnoea), and profound alterations in heart rate and blood pressure. These doses also caused salivation, lachrymation, skeletal muscle fasciculation and an elevation in body temperature. Concurrent increases in firing were recorded from the phrenic nerve and from respiratory and other skeletal muscles. It is concluded that atraxin produces the same syndrome in primates as that observed with whole milked male funnel-web venom.     

Venom glands and some associated muscles in sea snakes

The venom glands and related muscles of sea snakes conform in their general structure to those of the terrestrial elapids. The venom gland, however, is smaller in size and the accessory gland is considerably reduced. A similar pattern is found in the Australian elapid Notechis. The musculus compressor glandulae is well developed in the sea snakes and in some species its posterior-medial portion runs uninterruptedly from the origin to the insertion of the muscle. This might be considered as a primitive condition suggesting an early divergence of the sea snakes from an ancestral elapid stock. Three species of sea snakes, Aipysurus eydouxi, Emydocephalus annulatus, and E. ijimae, feed on fish eggs and have very small, but still functioning, venom glands. The reduced accessory gland of the sea snakes is apparently connected with their aquatic environment, as a similar condition is found also in the elapine Boulengerina annulata which lives in large lakes of Central Africa. The similarity in structure of the venom gland between sea snakes and Notechis scutatus may point to a possible phylogenetic relationship between this group of Australian elapids and hydrophiine snakes.     

Histology,histochemistry, and emptying mechanism of the venom glands of some elapid snakes

The venom glands of several species of elapid snakes are described. The main venom gland consists of many tubules which usually contain large amounts of secretion product. The accessory gland surrounds the entire venom duct and is usually composed of uniform mucous epithelium. The epithelium lining the tubules of the accessory gland of Naja naja is composed of two distinct types of cells. Histochemical tests indicate that the main venom gland reacts with mercury bromphenol blue and PAS but not with alcian blue. The accessory gland reacts with PAS and alcian blue, and not with mercury bromphenol blue. Treatment of sections with sialidase demonstrates the presence of a sialomucin in the accessory gland. Stimulation of the muscles associated with the venom gland offers an indication of the venom expulsion mechanism of Bungarus caeruleus. A comparison of the venom apparatus of elapid and viperid snakes emphasizes marked differences in the internal anatomy of the venom glands, muscles associated with the gland, and arrangement of glandular components. The morphological differences and dissimilar venom expulsion mechanisms support the recent view of the polyphyletic origin of venomous snakes.     

Intramuscular pressure, force and blood flow in rabbit tibialis anterior muscles during single and repetitive contractions

The elevated intramuscular pressure (IMP) associated with sustained muscle contraction can affect blood flow, and could influence the long-term viability of functional skeletal muscle grafts. We therefore examined the relationship between force, peak IMP and blood flow in the tibialis anterior muscle of the anaesthetized rabbit. During isometric contractions, IMP was related linearly to force, and only the slope of the relationship varied between animals. During isotonic contractions, however, the highest values of IMP were found at the lowest force levels, and IMP appeared to be related to the amount and speed of shortening. During repeated isometric contractions, the ratio of IMP to force varied with time, stimulation pattern and subject. Mean blood flow did not differ appreciably between␣repetitive isometric contractions at duty cycles of 10–40%, and was unrelated to integrated pressure, integrated force, or depth from the surface. We conclude: (1) that IMP is unlikely to affect mean blood flow during cyclic activity that has a duty cycle less than 40%; and (2) that the clinical use of IMP as a predictor of muscle force appears to be justified only for single isometric contractions, and needs to be interpreted cautiously when contractions involve shortening or fatigue. Accepted: 17 November 1997     

Morphology and variability of masticatory structures in the orangutan

The present study examines and describes variability and ontogenetic change in the masticatory structures of the orangutan. Mm. temporalis, masseter, pterygoideus lateralis, and pterygoideus medialis as well as m. digastricus are dissected, described, and removed unilaterally for drying from a sample of 10 orangutans. The temporomandibular joint and related structures are also described. Results indicate substantial interindividual variability in muscle morphology as well as possible age-related changes. Age-related changes in m. temporalis include an expansion of attachment area, adjustments in fiber direction, and changes in related bony structures. An increased attachment area is likewise seen in m. masseter in older individuals. Age-related variation is also seen in the position and extent of the origin of m. digastricus as well as in its fiber direction. A comparison of muscle weights indicates that m. pterygoideus lateralis is relatively larger in the neonate, whereas in the older individuals, it is relatively smaller compared to other masticatory muscles. The general morphology of the masticatory structures in the orangutan resembles that of other extant great apes and humans with the exception of m. digastricus. The orangutan differs in possessing only a posterior belly of this muscle.     

Effects of fibre type and diffusion distance on mouse skeletal muscle glycogen content in vitro

In vitro incubation of isolated rodent skeletal muscle is a widely used procedure in metabolic research. One concern with this method is the development of an anoxic state during the incubation period that can cause muscle glycogen depletion. Our aim was to investigate whether in vitro incubation conditions influence glycogen concentration in glycolytic extensor digitorum longus (EDL) and oxidative soleus mouse muscle. Quantitative immunohistochemistry was applied to assess glycogen content in incubated skeletal muscle. Glycogen concentration was depleted, independent of insulin‐stimulation in the incubated skeletal muscle. The extent of glycogen depletion was correlated with the oxidative fibre distribution and with the induction of hypoxia‐induced‐factor‐1‐alpha. Insulin exposure partially prevented glycogen depletion in soleus, but not in EDL muscle, providing evidence that glucose diffusion is not a limiting step to maintain glycogen content. Our results provide evidence to suggest that the anoxic milieu and the intrinsic characteristics of the skeletal muscle fibre type play a major role in inducing glycogen depletion in during in vitro incubations. J. Cell. Biochem. 107: 1189–1197, 2009. © 2009 Wiley‐Liss, Inc.     

Neuromuscular organization of avian flight muscle: Morphology and contractile properties of motor units in the pectoralis (pars thoracicus) of pigeon (Columba livia)

We used acid digestion and glycogen depletion to determine fascicle organization, fiber morphology, and physiological and anatomical features of individual motor units of an in-series muscle, the pectoralis (pars thoracicus) of the pigeon (Columba livia). Most fascicles are attached at one end to connective tissue. Average fiber length in the four regions examined range from 42% to 66% of average fascicle length. More than 65% of fibers are blunt at one end of a fascicle and taper intrafascicularly. Fibers with blunt–blunt endings range from 13% to 31% of the population in different regions; taper–taper fibers range from 2% to 17%. Pigeon pectoralis fibers are distinguished histochemically into fast-twitch glycolytic (FG) and fast-twitch oxidative-glycolytic (FOG) populations. Three units composed of FG fibers (FG units) contract more quickly than three units composed of FOG fibers (FOG units) (range 31–37 vs 47–62 msec), produce more tetanic force (0.11–0.32 vs 0.02–0.05 N) and are more fatigable (<18% initial force vs >50% after repeated stimulation). Most motor units are confined to one of the four muscle regions. Territory of two FOG units is <30% of parent fascicle length. Territories of other units spanned parent fascicles; most fibers in these units do not extend the full fascicle length. Compared to FG units, FOG units have lower maximum innervation ratios and density indices (ratio of depleted/total FOG fibers in territory 8–14% vs 58–76% for FG units). These differences support the hypothesis that FG units are organized to produce substantial force and power for takeoff, landing and other ballistic movements whereas FOG units are suited for sustained flight when power requirements are reduced. Implications of findings for understanding the control of in-series muscles and the use of connective tissue elastic elements during wing movements are discussed. J.Morphol. 236:179–208, 1998. © 1998 Wiley-Liss, Inc.     

The evolution of venom-delivery systems in snakes

The Colubroidea represents approximately 2300 of the 2700 species of living snakes and includes all venomous taxa. Although many morphological studies of colubroid snakes have been carried over the last hundred years, the phylogenetic relationships within this group are poorly known. In this study, components of the venom-delivery system (VDS) were examined within the context of two conflicting phylogenetic hypotheses proposed in 1988 by Cadle and in 1998 by Kraus & Braun. The results suggest that several major morphological changes occurred early in colubroid evolution: a Duvernoy's gland evolved, the posterior maxillary teeth became specialized relative to the anterior maxillary teeth, and the attachment of the pterygoideus muscle moved forward to a position associated with the posterior maxillary teeth. These innovations may have allowed the great radiation of colubroid snakes that led to the Colubroidea representing such a large percentage of living snakes. More recently, three separate lineages of colubroids have independently evolved highly specialized front-fanged VDSs with large and complex venom glands, venom gland compressor muscles, and tubular fangs.  © 2003 The Linnean Society of London, Zoological Journal of the Linnean Society, 2003, 137 , 337−354.     

Pterygoideus muscles and other jaw adductors in amphibians and reptiles

The general structural patterns of jaw adductors in all orders of extant amphibians and reptiles, and also polypteriforms, crossopterygians (coelacanth), and dipnoans, are compared. The pterygoideus muscles probably developed independently and in parallel in gymnophions and amniotes from the profound pseudotemporalis muscle, which was present in their fishlike ancestors and was retained in caudate and anuran amphibians. The functional causes of the development of pterygoideus muscles in the majority of tetrapod groups and the absence of these muscles in Urodela and Anura are discussed. The anterior pterygoideus muscle of crocodiles is homologous to the pseudotemporalis (superficial) muscle of other reptiles.     

How important is endogenous muscle glycogen to fatigue in prolonged exercise?

Endogenous muscle glycogen represents a primary fuel source during large muscle group activity in the human. The depletion of this fuel source during submaximal exercise at intensities ranging between 60 and 85% of maximal aerobic power (Vo2max) is widely believed to be the cause of an inability to sustain exercise. Alterations of preexercise muscle glycogen reserves by dietary and exercise manipulations and changing the degree of dependency on endogenous glycogen during exercise by modifying the availability of other fuel sources have in general served to establish a close relationship between muscle glycogen and fatigue resistance. However, in spite of the evidence implicating glycogen depletion to fatigue, the mechanism remains elusive. The most popular theory is that glycogen is an essential substrate, the depletion of which results in a reduction in the rate of ATP regeneration and an inability to maintain energy supply to one or more of the processes involved in excitation and contraction in the muscle. As a consequence, the muscle is unable to translate the motor drive into an expected force and fatigue develops. However, there is little experimental evidence to support this theory. Most studies report no or only minimal changes in ATP concentration at fatigue with low glycogen and no further change in the by-products of ATP hydrolysis. These findings suggest that fatigue might be caused by other nonmetabolic factors. This review examines these other nonmetabolic factors and analyzes their potential role in fatigue during prolonged exercise with depletion of muscle glycogen reserves.     

The effects of swimming and running regimens on skeletal muscle glycogen in the rat.

Endurance capacity and the effects of different post-exercise states on skeletal muscle glycogen have been studied in rats trained by swimming or running and in sedentary controls. Regular endurance exercise resulted in increased skeletal muscle glycogen stores. A greater depletion was observed in trained animals than in non-trained animals after a training bout or exhaustive exercise. While muscle glycogen levels did not reflect a differential training stimulus (running vs swimming), swimming as a measure of exhaustive exercise was deemed invalid because of the ability of trained swimmers to avoid stenuous exercise by an alteration of swimming pattern.     

Characterization of beta-adrenoceptors responsible for venom production in the venom gland of the snake Bothrops jararaca

We have shown that the stimulation of beta-adrenoceptors is an important step in venom production in the Bothrops jararaca venom gland. In the present study, the pharmacological profile of the beta-adrenoceptor present in Bothrops jararaca venom gland was characterized by radioligand binding assay and by the ability of isoprenaline to promote accumulation of cyclic AMP in dispersed secretory cells. In both cases, the venom glands were obtained from non-extracted snakes (quiescent stage) or from snakes which venom was extracted 4 days before sacrifice (venom production stimulated stage). [125I]-iodocyanopindolol ([125I]-ICYP) bound to extracted gland membranes in a concentration-dependent and saturable manner, but with low affinity. Propranolol, beta1- or beta2-selective adrenoceptors ligands displaced the [125I]-ICYP binding with low affinity, while selective beta3-adrenoceptor ligands did not displace the [125I]-ICYP binding. The displacement of [125I]-ICYP by propranolol was similar in non-extracted and extracted glands, showing the presence of beta-adrenoceptors in both stages. In dispersed secretory cells of non-extracted glands, isoprenaline (1 microM) increased the cyclic AMP production and propranolol (10 microM) was able to block this effect. On the other hand, in extracted glands, isoprenaline had no effect. The results suggest that the beta-adrenoceptors present in the Bothrops jararaca venom glands are different from those (beta1, beta2 or beta3) described in mammals, but are coupled to the Gs protein, like the known beta-adrenoceptor subtypes. Moreover, previous in vivo stimulation of venom production desensitizes the beta-adrenoceptors system and, although the receptors could be detected by binding studies, they are not coupled to the Gs protein, indicating that beta-adrenoceptors stimulation contributes to the initial steps of venom synthesis.     

Absolute strength of the muscles attached to the mandible

In the course of an anatomical investigation of the muscles, securing movements of the mandible, performed on 10 human corpses, the muscle fibre length, volume and weight of each muscle has been estimated. Owing to the formula suggested by P. F. Leshaft (1880)--q = v/e, the physiological diameter of the muscles has been determined. Since the muscle with the diameter equal to 1 cm2 develops an absolute forse of 10 kg, the absolute muscle force value of the anterior group of muscles has been obtained for the first time (venter anterior musculi digastri--4.8 kg, musculus mylohyoideus--10.7 kg and musculus geniohyoideus--6.3 kg). The data on the absolute force of the posterior group of muscles has been verified (musculus masseter--24.2 kg, musculus temporalis--28 kg, musculus pterygoideus medialis--15.6 kg and musculus pterygoideus lateralis--15.5 kg). Analysing the interaction of forces of the muscles participating in the mandible movements, direction and value of displacements of the mandibular fragments have been explained and confirmed on some clinical examples. The data on the absolute force of the muscles studied can be used for investigating the displacement mechanism of the mandibular fragments after its resection and when its integrity is broken.