Electrophysiological properties of biventer cervicis muscle fibers of normal and roller pigeons.

Cable parameters, excitability characteristics, and contractile response to acetylcholine were measured in biventer cervicis muscles from Helmet pigeons, Racing Homer pigeons and Parlor (nonflying) Roller pigeons. Cable parameters for the three strains, were respectively: calculated diameter, 30.1, 42.5, and 37.3 mum; membrane resistance, 450, 556, and 386 omega-cm2; membrane capacitance, 4.2, 3.9, and 4.5 muF/cm2, and myoplasmic resistivity, 79, 185, and 116 omega-cm. Significant differences between excitability characteristics of Homer pigeon and Roller pigeon fibers were a 17% shorter maximal latency for spike initiation (P less than 0.025) and 24% lower rheobasic current (P less than 0.05) in Roller fibers. Dose-response curves of isolated biventer cervicis to acetylcholine revealed slight, but significant, differences between Helmets and Rollers. These are the first electrophysiological data from pigeon skeletal muscle and the first from any avian biventer cervicis. The biventer muscles of chickens contain mainly "slow" fibers, but our results show that pigeon biventer fibers have properties similar to the "fast" PLD fibers of the chicken. Furthermore, the existence of different myoplasmic resistivities for each strain of pigeons used in this study suggests the need for more careful determination of this parameter in electrophysiological investigations. Although our results show that Roller pigeon fibers differ from those of nonrolling pigeons in the respects described above, these differences are minor in comparison to the severe behavioral abnormalities of Roller pigeons. Some yet untested component of neuromuscular transmission may be directly involved in the rolling phenomenon, but the differences we report may simply be due to strain differences, muscle hypertrophy, or a more severe defect elsewhere in the nervous system.     

Studies on action mechanisms of a possible false cholinergic transmitter, (2-hydroxyethyl) methyldiethylammonium

(2-Hydroxyethyl) methyldiethylammonium (DEC; Diethylcholine) was found to inhibit cholinergic fibers slowly, both in skeletal muscle (ED₅₀: 2.25 × 10^?5 M in chick biventer cervicis and 42 mg/kg in rat sciatic-gastrocnemius) and in smooth muscle preparations (ED₅₀: 7.7 × 10^?4 M in transmurally stimulated guinea-pig ileum) without having any effect on dose-response curves of acetylcholine to contract chick biventer cervicis, frog rectus abdominis and guinea-pig ileum. These results indicate that DEC acts at the prejunctional nerve fibers, but not at the postjunctional cholinergic receptor sites. DEC was acetylated efficiently both by choline acetyltransferase and by minced rat brain, suggesting that it can be acetylated to acetyl-DEC in the nerve ending. Acetyl-DEC was found to block acetylcholine actions competitively both in smooth and in skeletal muscle preparations (1 × 10^?3 ? 1 × 10^?2M) indicating that the acetylated product of DEC can serve as an antagonist at the cholinergic receptor site. It is therefore concluded that DEC is a false cholinergic transmitter.     

The functional organization and motor innervation of the cervical skeletal muscle in mammals

Structural and functional studies have been done on the nerve-muscle transmission in m. biventer cervicis of mouse using electrophysiological, mechanographic and microscopic techniques. It was shown that each muscle fiber has several synaptic inputs, although m. biventer cervicis is a typical phasic skeletal muscle. Usually, there is a single large synapse of the end-plate type and several small synaptic contacts. Some of the synapses investigated produce typical end-plate potential which is transformed into the action potential. Certain part of synapses are capable of producing action potentials only during facilitation of the transmission by rhythmic stimulation. In curarized preparations, supramaximum single indirect stimulus may evoke in a single point of a fiber multicomponent postsynaptic responses of various amplitude and, sometimes, of various latency. Contractile response of a single segment to a maximum direct stimulation is always higher than to an indirect one. Unusual functional properties of m. biventer cervicis are discussed in relation to peculiarities of its origin.     

Neuromuscular action of venom from the South American colubrid snake Philodryas patagoniensis

Snakes of the opisthoglyphous genus Philodryas are widespread in South America and cause most bites by colubrids in this region. In this study, we examined the neurotoxic and myotoxic effects of venom from Philodryas patagoniensis in biventer cervicis and phrenic nerve-diaphragm preparations and we compared the biochemical activities of venoms from P. patagoniensis and Philodryas olfersii. Philodryas patagoniensis venom (40 microg/mL) had no effect on mouse phrenic nerve-diaphragm preparations but caused time-dependent neuromuscular blockade of chick biventer cervicis preparations. This blockade was not reversed by washing. The highest concentration of venom tested (40 microg/mL) significantly reduced (p<0.05) the contractures to exogenous acetylcholine (55 microM and 110 microM) and K(+) (13.4 mM) after 120 min; lower concentrations of venom had no consistent or significant effect on these responses. Venom caused a concentration- and time-dependent release of creatine kinase (CK) from biventer cervicis preparations. Histological analysis showed contracted muscle fibers at low venom concentrations and myonecrosis at high concentrations. Philodryas venoms had low esterase and phospholipase A(2) but high proteolytic activities compared to the pitviper Bothrops jararaca. SDS-PAGE showed that the Philodryas venoms had similar electrophoretic profiles, with most proteins having a molecular mass of 25-80 kDa. Both of the Philodryas venoms cross-reacted with bothropic antivenom in ELISA, indicating the presence of proteins immunologically related to Bothrops venoms. RP-HPLC of P. patagoniensis venom yielded four major peaks, each of which contained several proteins, as shown by SDS-PAGE. These results indicate that P. patagoniensis venom has neurotoxic and myotoxic components that may contribute to the effects of envenoming by this species.     

Isolation and purification of a lethal scorpion toxin with neuromuscular blocking activity

Toxin-L a lethal neuromuscular blocking agent was isolated from the venom of the scorpion, Lychas laevifrons (Pocock), by the CM-cellulose ion-exchange chromatography. It was a homogenous, thermolabile and low molecular weight protein. The toxin produced irreversible blockade of indirect stimulation induced twitch responses on innervated rat phrenic nerve-diaphragm and chick biventer cervicis preparation. The toxin did not produce any contractile response on toad rectus abdominis muscle preparation. On chronically denervated rat diaphragm, the toxin failed to alter the responses induced by direct stimulation, exogenous acetylcholine, potassium chloride and caffeine. Acetylcholine and carbachol induced contractions on isolated chick biventer cervicis remained unaltered by the toxin. Neostigmine failed to alter toxin induced neuromuscular blockade on innervated rat diaphragm. The toxin released a significant amount of acetylcholine from innervated rat diaphragm. It may be concluded that the toxin acts presynaptically through the release of acetylcholine, thereby producing neuromuscular blockade.     

Aging alters contractile properties and fiber morphology in pigeon skeletal muscle

In this study, we tested the hypothesis that skeletal muscle from pigeons would display age-related alterations in isometric force and contractile parameters as well as a shift of the single muscle fiber cross-sectional area (CSA) distribution toward smaller fiber sizes. Maximal force output, twitch contraction durations and the force–frequency relationship were determined in tensor propatagialis pars biceps muscle from young 3-year-old pigeons, middle-aged 18-year-old pigeons, and aged 30-year-old pigeons. The fiber CSA distribution was determined by planimetry from muscle sections stained with hematoxylin and eosin. Maximal force output of twitch and tetanic contractions was greatest in muscles from young pigeons, while the time to peak force of twitch contractions was longest in muscles from aged pigeons. There were no changes in the force–frequency relationship between the age groups. Interestingly, the fiber CSA distribution in aged muscles revealed a greater number of larger sized muscle fibers, which was verified visually in histological images. Middle-aged and aged muscles also displayed a greater amount of slow myosin containing muscle fibers. These data demonstrate that muscles from middle-aged and aged pigeons are susceptible to alterations in contractile properties that are consistent with aging, including lower force production and longer contraction durations. These functional changes were supported by the appearance of slow myosin containing muscle fibers in muscles from middle-aged and aged pigeons. Therefore, the pigeon may represent an appropriate animal model for the study of aging-related alterations in skeletal muscle function and structure.     

Absence of changes in fiber type with aging in avian muscles

Summary The anterior latissimus dorsi of the pigeon, a slow tonic muscle, and biventer cervicis, a mixed muscle, of two age groups (1–2 years old versus 6–8 years old) were compared with respect to percentages of fiber types and activities of adenosine triphosphatase (ATPase) and succinic dehydrogenase (SDH), as estimated histochemically, to determine whether these became altered with old age. These parameters did not change between the young and old birds for either type of muscle.This investigation was supported in part by research grants PCM 77-15960 and PCM 79-16540 from the National Science Foundation     

Oscillation in resting tension of chick skeletal muscle treated with toxin V from scorpion venom, reduced [calcium]o and cadmium

1. An oscillation in resting tension of the chick biventer cervicis muscle is induced by scorpion toxin (CsT-V), reduced [Ca]o and by Cd2+. Other chick muscles, e.g. semispinalis cervicis, anterior and posterior latissimus dorsi, also oscillate with the same treatments. The period of the oscillation in the biventer cervicis muscle is uniform, averaging 5-6 min. 2. The oscillation and the period are very sensitive to changes in temperature, [Ca]o, [K]o, ouabain and tetrodotoxin [TTX]. The oscillation is resistant to twitch blocking dosages of nicotinic antagonists. 3. The period of the oscillation varies inversely as the temperature (26-37 degrees C), log [K]o and [ouabain] and directly as the log [Ca]o and [TTX]. 4. These data support the inference that the oscillator is postjunctional and results from fluctuation in the electrochemical gradient of ions across the muscle cell membrane. The role, if any, of the Na+, K+ pump in the primary oscillogenic mechanism is not evident from our data.     

Reversible neuromuscular blocking action of carboxymethylated alpha-bungarotoxin

Carbamidomethylation of the reduced Cys29-Cys33 bridge of alpha-bungarotoxin (Bungarus multicinctus postsynaptic neurotoxin) did not alter the LD50 or irreversibility of the toxin, while carboxymethylated alpha-bungarotoxin was less potent and its neuromuscular blocking action in mouse diaphragm was reversible. The circular dichroic spectra of both modified toxins were similar but slightly different from that of native toxin. Neuromuscular transmission in the chick biventer cervicis nerve-muscle preparation could be blocked by the carboxymethylated toxin, and reactivated by washing, whereas the response of the muscle to extrinsic acetylcholine could also be blocked but was hardly restored by washing. These results suggest that carboxymethylated toxin can differentiate between junctional and extrajunctional acetylcholine receptors in chick skeletal muscle.     

Reversible neuromuscular blocking action of carbohymethylated α-bungarotoxin

Carbanidomethylation of the reduced Cys₂₉-Cys₃₃ bridge of α-bungarotoxin (Bungarus multicinctus postsynaptic neurotoxin) did not alter the LD₅₀ or irreversibility of the toxin, while carboxymethylated α-bungarotoxin was less potent and its neuromuscular blocking action in mouse diaphragm was reversible. The circular dichroic spectra of both modified toxins were similar but slightly different from that of native toxin. Neuromuscular transmission in the chick biventer cervicis nerve-muscle preparation could be blocked by the carboxymethylated toxin, and reactivated by washing, whereas the response of the muscle to extrinsic acetylcholine could also be blocked but was hardly restored by washing. These results suggest that carboxymethylated toxin can differentiate between junctional and extrajunctional acetylcholine receptors in chick skeletal muscle.     

Effect of phospholipase A on actions of cobra venom cardiotoxins on erythrocytes and skeletal muscle

The actions of two phospholipase-free cardiotoxins from the venom of the cobra Naja naja siamensis were compared to phospholipase-contaminated cardiotoxins in terms of their ability to lyse human erythrocytes and to depolarize and contract skeletal muscle. The presence of 3–5% (w/w) phospholipase caused a 20–30-fold increase in the haemolytic activity of the two cardiotoxins, the pure cardiotoxins being virtually without haemolytic activity at 10^?7-10^?6 M. Phospholipase contamination did not enhance the ability of the cardiotoxins to cause contracture of chick biventer cervicis muscles and it caused less than a 2-fold increase in the depolarizing activity of the cardiotoxins on cultured skeletal muscle. Phospholipase-free cardiotoxins were about 10–20-times more active on cultured skeletal muscle fibres than on erythrocytes. These results support the hypothesis that some cardiotoxins have more affinity for the membranes of excitable cells than for those of other cells such as erythrocytes.     

Pharmacological actions of tentacle extract of the jellyfish, Acromitus rabanchatu, occurring in the Bay of Bengal

Tentacle extract of A.rabanchatu, produced a fall of blood pressure in cat, rat and guinea pig. Hypotension produced in cat remained unantagonized by blockers of acetylcholine, histamine and 5-HT. On isolated guinea pig heart, the extract significantly reduced the rate and amplitude of contraction leading to irreversible cardiac arrest. In cats and rats, the respiratory rate and amplitude was decreased significantly and resulted in temporary apnoea. The extract also produced vasoconstriction in perfused rat hindquarter preparation and increased cutaneous capillary permeability. The extract produced contraction in several isolated smooth muscle preparations. Contraction on guinea pig ileum was partly antagonized by atropine and cyproheptadine. On isolated rat phrenic nerve diaphragm and chick biventer cervicis, the extract produced irreversible blockade of the electrical stimulation-induced twitch responses. Haemolytic and myonecrotic activity was exhibited by the extract. LD50 was found to be 7.7 mg/kg (iv, mice).     

Conduction velocities and fiber diameters in a cutaneous nerve of the pigeon

Summary The characteristics of fibers of a cutaneous nerve supplying the wing skin of the pigeon have been investigated with electrophysiological and electron microscopic techniques.Recordings of the compound action potential showed four distinct peaks with conduction velocities of about 30 m/s, 12 m/s, 4 m/s and 0.5 m/s.From electron micrographs both fiber diameters and thickness of myelin sheath were assessed and used as criteria for segregating various fiber populations. Altogether four groups could be discerned: large thickly myelinated fibers, small thickly myelinated fibers, small thinly myelinated fibers, and unmyelinated or C-fibers. The subdivision of the thickly myelinated fibers into two populations is evidenced mainly by corresponding peaks in the compound action potential. The thinly myelinated fibers with a mean diameter of 2 m contributed about 90% of all myelinated fibers in this nerve.When comparing fiber dimensions and conduction velocities of this avian nerve with those of mammalian cutaneous nerves, the lower CV's of avian nerve fibers can be explained by smaller diameters and thinner myelin sheaths.The results of this investigation are a prerequisite for latency considerations in central somatosensory pathways in birds.Abbreviations CAP compound action potential - CV conduction velocity - D fiber diameter - d axon diameter - g ratio d/D - m thickness of myelin sheath     

Tenotomy of the avian anterior latissimus dorsi muscle. II. Can regeneration from the stump occur in the pigeon?

Because the chick's anterior latissimus dorsi muscle (ALD) regenerates a fast-twitch muscular connection after tenotomy, the pigeon's ALD was tenotomized, either at the origin or through the muscle 0.5 cm from the origin, to determine whether this muscle behaves similarly to the chick muscle. These procedures were compared in pigeons operated upon at 7 weeks, versus 5 to 9 months of age. The pigeon's ALD did not regenerate a new connection, and other differences were observed between the pigeon and chick ALD. The pigeon ALD has only a single slow muscle-fiber type, has fewer fast fibers, and transforms to a fast-twitch muscle more readily than the chick ALD after tenotomy. The transformation of muscle fiber types occurred more readily in the older pigeons than those tenotomized at 7 weeks of age. Tenotomy induced morphological alterations of the muscle fiber structure in all of the pigeons, which is in contrast to the absence of changes in the tenotomized chick ALD. Therefore the pigeon and chick ALD respond completely differently to tenotomy.     

Shivering thermogenesis in leg and breast muscles of galliform chicks and nestlings of the domestic pigeon.

We studied the ontogeny of shivering thermogenesis in breast and leg muscles of precocial galliforms (domestic fowl, grey partridge, and Japanese quail) and the altricial domestic pigeon using electromyography (EMG) and indirect calorimetry. Galliforms were able to increase heat production by shivering in leg muscles at the youngest age studied (1-2 d). Pectorals contributed to heat production from days 7-10 onward, but in the partridge and especially in the fowl, shivering by the pectorals was weaker than in the quail. In the pigeon, shivering began in pectorals and legs at 2 and 4 d of age, respectively, and pectorals had clearly the predominant role in thermogenesis. Despite the early beginning of electrical signs of shivering, significant thermogenesis did not appear in the pigeon before the age of 6 d. All galliforms shivered in bursts, like pigeons aged 2-4 d. From the age of 6 d onward, continuous shivering became predominant in the pigeon. In pectorals of 2-6-d-old pigeons, shivering did not increase linearly during decreasing ambient temperature, as in other muscles and species, but started abruptly, at full intensity. Furthermore, in 2-4-d-old pigeons, cooling induced movement activity in legs. The median frequency of shivering EMGs varied (1) with maturation of the muscle, (2) with size of the adult bird, and (3) between altricials and precocials.     

Probability of Detecting Band-Tailed Pigeons During Call-Broadcast Versus Auditory Surveys

Abstract: Estimates of population trend for the interior subspecies of band-tailed pigeon (Patagioenas fasciata fasciata) are not available because no standardized survey method exists for monitoring the interior subspecies. We evaluated 2 potential band-tailed pigeon survey methods (auditory and call-broadcast surveys) from 2002 to 2004 in 5 mountain ranges in southern Arizona, USA, and in mixed-conifer forest throughout the state. Both auditory and call-broadcast surveys produced low numbers of cooing pigeons detected per survey route (x̄ ≤ 0.67) and had relatively high temporal variance in average number of cooing pigeons detected during replicate surveys (CV ≥ 161%). However, compared to auditory surveys, use of call-broadcast increased 1) the percentage of replicate surveys on which ≥1 cooing pigeon was detected by an average of 16%, and 2) the number of cooing pigeons detected per survey route by an average of 29%, with this difference being greatest during the first 45 minutes of the morning survey period. Moreover, probability of detecting a cooing pigeon was 27% greater during call-broadcast (0.80) versus auditory (0.63) surveys. We found that cooing pigeons were most common in mixed-conifer forest in southern Arizona and density of male pigeons in mixed-conifer forest throughout the state averaged 0.004 (SE = 0.001) pigeons/ha. Our results are the first to show that call-broadcast increases the probability of detecting band-tailed pigeons (or any species of Columbidae) during surveys. Call-broadcast surveys may provide a useful method for monitoring populations of the interior subspecies of band-tailed pigeon in areas where other survey methods are inappropriate.     

Muscle insensitivity to tetrodotoxin: Induction by α-bungarotoxin and removal by submechanical threshold stimulation

Intramuscular injection of alpha-bungarotoxin (αBGT) into rat extensor digitorum longus muscles produced pharmacological blockade of neuromuscular transmission and resulted in denervation-like changes (Berg, D. and Hall, Z. (1975), J. Physiol. (London) 244: 659–676). More than 80% of fibers from αBGT-injected muscles produced action potentials (ap's) in the presence of tetrodotoxin (TTX, 1 × 10^?6 M). Chronic electrical stimulation of these muscles, below the level necessary to elicit a contraction, resulted in a marked return toward normal of sarcolemmal sensitivity to TTX. After 4 days of submechanical threshold stimulation, less than 45% of αBGT-injected fibers produced ap' in the presence of TTX, whereas more than 80% of unstimulated fibers from contralateral control muscles exhibited resistance to TTX. These findings indicate that in addition to sarcolemmal sensitivity to acetylcholine, sensitivity of the sodium conductance mechanism to TTX is also directly influenced by muscle activity independent of contraction.     

Effects of 5-hydroxytryptophan on serotonin in nerve endings

—Preparations of synaptosomes (P₂) from the telencephalon and from the diencephalon plus optic lobes of the pigeon and from the telencephalon of the rat were used to study the effects of 5-hydroxytryptophan (5-HTP) on (a) the levels of serotonin (5-HT) in nerve endings and (b) the release of 5-HT from nerve endings. The levels of 5-HT were significantly higher (3.21 × 0.35 nmol/g original tissue weight) in the P₂ fraction isolated from the telencephalon of pigeons given intramuscular injections of 50mg/kg of d ,l -5-HTP in comparison to control values (1.42 ± 0.07). A similar twofold increase was observed with the P₂ fraction isolated from the diencephalon plus optic lobes. In addition, the levels of 5-HTP and 5-hydroxyindoleacetic acid also increased significantly in these P₂ fractions isolated from pigeons given d ,l -5-HTP injections in comparison to values obtained for pigeons given saline injections. In vitro studies using preparations of synaptosomes (from both pigeon and rat) labelled with [³H]5-HT indicated that 0.10 mil l -5-HTP increased the release of [³H]5-HT twofold over control values. A concentration as low as 0.001 mm l -5-HTP was tested on the P₂ fraction from the telencephalon of the pigeon and was found to significantly increase the release of [³H]5-HT over control values. This effect by l -5-HTP was blocked if a decarboxylase inhibitor was added to the medium. l -5-HTP at a concentration of 1.5 mm had no apparent effect on the release of [³H]norepinephrine or [³H]dopamine from synaptosomes prepared from the telencephalon of the rat or pigeon. The results are discussed in terms of the role of serotonin in producing certain types of behavioral depressions exhibited by pigeons and rats given injections of 5-HTP.     

Parasites from feral pigeons as a health hazard for humans

Feral pigeons live in almost every large city in the world. The feeding of pigeons by humans, deliberately or accidentally, allows the build‐up of large populations that can cause a variety of problems, primarily the fouling of buildings and monuments by their droppings. Excreta of wild birds are a well‐known source of pathogenic microorganisms that can cause infection in man. The most important ectoparasites of feral pigeons are the red blood mite, Dermanyssus gallinae, and the pigeon tick, Argas reflexus, both of which can migrate into human living space when they lose their natural hosts. The bites of the red blood mite are irritating but harmless, whereas the pigeon tick can cause severe health problems to predisposed persons. After repeated bites from A. reflexus, people can develop IgE‐mediated (type I) allergy, which in extreme situations may lead to life‐threatening symptoms of anaphylactic shock. So far, one fatality due to A. reflexus deriving from feral pigeons has been reported. In the case of parasitic infestations of humans, the source must be removed by excluding pigeons from further breeding and by disinfection of the environment of pigeon‐breeding sites.     

Differences in purity of commercial samples of hemicholinium-3: effects on high affinity choline uptake, neuromuscular transmission, acetylcholinesterase and carbachol and acetylcholine contractions.

Commercial samples of hemicholinium-3 (HC-3) have been found to vary in colour (from white to a sandy-yellow colour) and chemical composition. There were no significant differences between the various HC-3 samples with regards to inhibition of high affinity choline uptake into synaptosomes or inhibition of neuromuscular transmission in the chick biventer cervicis (CBC) preparation. Yellow HC-3 inhibited acetylcholinesterase more than white HC-3 with I₅₀ of 4.8 × 10^?5 M and 3.3 × 10^?4 M, respectively. Carbachol-induced contractions of the CBC preparation were inhibited more by yellow HC-3 than white HC-3; the opposite was true for acetylcholine- induced contractions. The results indicated that there is a minor contaminant in yellow HC-3 other than deanol which was a potent inhibitor of acetylcholinesterase and the carbachol response.