Latrodectus Mactans Tredecimguttatus Research Articles

Une paralysie inhabituelle : piqûre par la veuve noire

Introduction : The mediterranean black widow spider is a kind of latrodectus mactans tredecimguttatus. It is found in the mediterranean bassin. Its bite causes a rare syndrome called lactrodectism. Observation : A moroccan patient, aged 17 years old, came to the emergency ward with severe pain in the right chest and fever, insect bites scar at the elbow. The evolution was favorable with antibiotics, antihistamines, and intravenous calcium. The discovery by the family in the patient’s room of the spider identified type Latrodectus has consolidated the diagnosis of spider bite. Discussion : Latrodectus tredecimguttatus is a spider that can be responsible of envenomation in humans. Its bite is almost painless. The α-neurotoxin protein latrotoxine is implicated in the onset of symptoms. this symptomatology is polymorphic, including a latrodectismica facies, with or without constitutional symptoms, painful symptoms, cardiovascular signs and digestive signs that represent the the most common pattern of consulting and most often a source of diagnostic errors. Latrodectism is a source of frequent misdiagnosis, surgical acute abdomen, colic, tetanus, meningitis, psychosis, myocardial infarction or opiate withdrawal syndrome inducing inappropriate treatment, often detrimental to the poisoned patient. Conclusion : This poisoning, although rare, must be known because it can lead to unnecessary therapeutic procedures, and even be potentially fatal for extremely aged people with extreme ages of life.

Does extracellular calcium determine what pool of GABA is the target for α-latrotoxin?

Presynaptic neurotoxin α-latrotoxin, from the venom of Latrodectus mactans tredecimguttatus, causes massive [ 3 H ]GABA release from rat brain synaptosomes, irrespective of calcium presence in the extracellular medium. Whether the binding of α-latrotoxin to Ca 2+-dependent (neurexin 1α) or to Ca 2+-independent (latrophilin) receptor triggers [ 3 H ]GABA release by the same mechanisms or different ones, inducing either exocytotic process or outflow by mobile membrane GABA transporter, is unknown. We examined α-latrotoxin-evoked [ 3 H ]GABA release from synaptosomes which cytosolic [ 3 H ]GABA pool was depleted either by applying competitive inhibitors of the GABA transporter, nipecotic acid and 2,4-diaminobutyric acid, or by permeation with digitonin. We also compared the effect of the GABA transporter inhibitors on depolarisation-evoked and α-latrotoxin-evoked [ 3 H ]GABA release using as depolarising agents 4-aminopyridine and high KCl in the Ca 2+-containing and in Ca 2+-free medium, respectively. Incubation of synaptosomes with nipecotic acid induced the essential acceleration of unstimulated [ 3 H ]GABA release and deep inhibition of high KCl-evoked Ca 2+-independent [ 3 H ]GABA release. In contrast, at the similar conditions the effect of α-latrotoxin was greatly augmented with respect to the control response. Another way to assay what GABA pool was involved in α-latrotoxin-induced release lays in an analysis of the effects of depolarisation and α-latrotoxin in consecutive order. The preliminary 4-aminopyridine-stimulated [ 3 H ]GABA release attenuated the toxin effect. But when depolarisation occurred in Ca 2+-free medium, no influence on α-latrotoxin effect was revealed. Employing digitonin-permeated synaptosomes, we have shown that α-latrotoxin could stimulate [ 3 H ]GABA release in the medium with 1 mM EGTA, this effect of the toxin was blocked by concanavalin A and was ATP-dependent. The latter suggests that α-latrotoxin-released neurotransmitter has the vesicular nature. We assume that the type of the toxin membrane receptor does not determine the mechanisms of [ 3 H ]GABA release evoked by α-latrotoxin.

Latrodectisme chez un enfant

The malmignatte Latrodectus mactans tredecimguttatus, commonly known as a black widow spider, can be found in the Mediterranean region. Its bite is a cause of a rarely seen syndrome called latrodectism. During a visit to Corsica, a 13-year-old boy developed abrupt severe abdominal pain and spasmodic muscular contractions, headache and vomiting. The patient was restless and experienced hallucinations including distressing visions of death. His high blood pressure (154/100 mmHg) returned to normal within 3 days. Clinical examination revealed dyspnea and facial edema associated with blepharoconjunctivitis and hyperreflexia, together with a scattered erythema and pruritus. These changes took place within minutes, after a probable black widow spider bite. The abdominal and neuropsychiatric symptoms disappeared after 5 days. Treatment with calcium gluconate, paracetamol, phloroglucinol and hydroxyzine had no effect, but diazepam decreased the acuteness of the symptoms. Anti-venom serum was not used. Diagnosis of latrodectism must be based on clinical and epidemiological data. Erroneously diagnosing surgical acute abdomen, renal colic, meningitis, tetanus or opioid withdrawal would entail incorrect treatment.

Probing the structure-function relationship of α-latrotoxin-formed channels with antibodies and pronase

The major toxic component of black widow spider (Latrodectus mactans tredecimguttatus) venom, α-latrotoxin, is known to form ionic channels in different membranes. In order to probe the extramembrane domains of α-latrotoxin molecule, α-latrotoxin channels in planar lipid membrane were treated with antibodies to latrotoxin or with pronase added to different sides of the membrane. It was found that antibody addition to the same side as the toxin ( cis) decreased channel conductance only at positive potentials across the membrane. In contrast, trans side addition of antibodies changed the channel conductance at both positive and negative potentials: at positive potential conductance first slightly increased then decreased by more then 50%; at negative potential it decreased much more quickly, to only about 20% of the initial value. No dependence on membrane potential was found for pronase treatment of incorporated channels. For both cis and trans application of pronase, channel selectivity for Ca 2+ , Mg 2+, Ba 2+ and K +, Na +, Li + ions did not change significantly but Cd 2+ block was decreased. Trans pronase treatment also resulted in some rectification of I/V curves and an increase in channel conductance. We interpret these findings as evidence that α-latrotoxin channel has protruding parts on both sides of the membrane and that its conformation in the membrane depends on membrane potential.

Cloning and Structure of δ-Latroinsectotoxin, a Novel Insect-specific Member of the Latrotoxin Family: FUNCTIONAL EXPRESSION REQUIRES C-TERMINAL TRUNCATION

The venom of the black widow spider (BWSV) (Latrodectus mactans tredecimguttatus) contains several potent, high molecular mass (>110 kDa) neurotoxins that cause neurotransmitter release in a phylum-specific manner. The molecular mechanism of action of these proteins is poorly understood because their structures are largely unknown, and they have not been functionally expressed. This study reports on the primary structure of delta-latroinsectotoxin (delta-LIT), a novel insect-specific toxin from BWSV, that contains 1214 amino acids. delta-LIT comprises four structural domains: a signal peptide followed by an N-terminal domain that exhibits the highest degree of identity with other latrotoxins, a central region composed of 15 ankyrin-like repeats, and a C-terminal domain. The domain organization of delta-LIT is similar to that of other latrotoxins, suggesting that these toxins are a family of related proteins. The predicted molecular mass and apparent mobility of the protein (approximately 130 kDa) encoded in the delta-LIT gene differs from that of native delta-LIT purified from BWSV (approximately 100 kDa), suggesting that the toxin is produced by proteolytic processing of a precursor. MALDI-MS of purified native delta-LIT revealed a molecular ion with m/z+ of 110916 +/- 100, indicating that the native delta-LIT is 991 amino acids in length. When the full-length delta-LIT cDNA was expressed in bacteria the protein product was inactive, but expression of a C-terminally truncated protein containing 991 residues produced a protein that caused massive neurotransmitter release at the locust neuromuscular junction at nanomolar concentrations. Channels formed in locust muscle membrane and artificial lipid bilayers by the native delta-LIT have a high Ca2+ permeability, whereas those formed by truncated, recombinant protein do not.

Neurotoxin from black widow spider venom. Structure and function.

As known some natural neurotoxins can effectively influence the secretion process of a neurotransmitter from the nerve ending. Among such neurotoxins particular interest is presented by the toxic components of the black widow spider Latrodectus mactans tre-decimguttatus. Latrodectus toxins affect on all synapse types of various classes of animals, causing a massive release of a transmitter from the nerve terminal (Longenecker et al., 1970; Frontali, 1972; Grasso et al., 1979). α-Latrotoxin (α-LTX) is major toxic component of the black widow spider venom selectively toxic to vertebrates. The toxin can induce the transmitter release from nerve terminals and PC cells both in calcium and calcium-free medium (Grasso, 1988). The cDNA encoding the putative α-LTX precursor contains the 4203 base-pair open reading frame corresponding to the 156,855 Da protein composed of 1401 amino acid residues (Kiyatkin et al., 1990). But the molecular mass of the toxin deduced from the cDNA considerably differs from that determined for α-LTX by means of SDS gel electrophoresis (Ushkarev & Grishin, 1986) assuming possible processing in the C-terminal part of the polypeptide chain during its maturation. The size of the α-LTX molecule as well as its multifunctional properties might testify to the existence of several functional domains in the toxin structure. The paper summarizes the experimental results, which allow us to define some functional domains of the α-LTX molecule.

Cloning and structural analysis of alpha-latroinsectotoxin cDNA. Abundance of ankyrin-like repeats.

alpha-Latroinsectotoxin (alpha-LIT), purified from venom glands of the black widow spider Latrodectus mactans tredecimguttatus, is a presynaptic neurotoxin selective only for insects. A cDNA encoding the putative alpha-LIT precursor was isolated from a spider venom gland cDNA library. The cDNA contains a 4236-base-pair open reading frame corresponding to a 157826-Da protein composed of 1411 amino acids. The mature alpha-LIT, with molecular mass approximately 130 kDa, is probably derived from double processing in the N-terminal and C-terminal regions of the primary translation product. The structure region, extending over residues 464-1176, is composed almost entirely of ankyrin-like repeats which represent a motif also found in the alpha-latrotoxin (alpha-LTX), which has selective action on vertebrates. Total alignment of the alpha-LIT and alpha-LTX amino acid sequences reveals an overall similarity of 34.1%. Strong sequence divergence is observed in analogous cysteine-rich regions situated within the ankyrin-repeat domains of both alpha-LIT and alpha-LTX.

Crustacean-specific neurotoxin from the black widow spider Latrodectus mactans tredecimguttatus venom and its interaction with plasmatic membranes from Paralithodes camtshatica crab nerve ganglia: O. G. Shamotienko (Shemyakin Institute of Bioorganic Chemistry, U.S.S.R. Academy of Sciences, Moscow, Russia)

Crustacean-specific neurotoxin from the black widow spider Latrodectus mactans tredecimguttatus venom and its interaction with plasmatic membranes from Paralithodes camtshatica crab nerve ganglia: O. G. Shamotienko (Shemyakin Institute of Bioorganic Chemistry, U.S.S.R. Academy of Sciences, Moscow, Russia)

Effects of Latrodectus spider venoms on sensory and motor nerve terminals of muscle spindles.

The effects of the venoms of the spiders Latrodectus mactans tredecimguttatus (black widow) and Latrodectus mactans hasselti (red back) on sensory nerve terminals in muscle spindles were studied in the mouse. A sublethal dose of venom was injected into tibialis anterior and extensor digitorum longus muscles of one leg. After survival from 30 minutes to 6 weeks muscles were examined in serial paraffin sections impregnated with silver or by electron microscopy. Sensory endings became swollen, some within 30 minutes, while over the next few hours there was progressive degeneration of annulospiral endings. By 24 hours every spindle identified by light or electron microscopy was devoid of sensory terminals. Degenerated nerve endings were taken up into the sarcoplasm of intrafusal muscle fibres. Regeneration of sensory axons began within 24 hours, new incomplete spirals were formed by 5 days and by 1 week annulospiral endings were almost all normal in appearance. Intrafusal motor terminals underwent similar acute degenerative and regenerative changes. These experiments show that intrafusal sensory and motor terminals are equally affected by Latrodectus venoms. Sensory nerve fibres possess a capacity for regeneration equal to that of motor fibres and reinnervate intrafusal muscle fibres close to their original sites of innervation.

Peripheral catecholamine release by alpha-latrotoxin in the rat.

Intraarterial injection of alpha-latrotoxin (alpha LTx), the major toxin of the black widow spider (Latrodectus mactans tredecimguttatus) venom, into carotid catheterized rats, induced prompt and marked rises in plasma adrenaline and noradrenaline concentrations, indicating that the toxin stimulates catecholamine release from both the adrenal medulla and sympathetic nerve terminals. Pretreatment with the ganglionic blocker chlorisondamine greatly reduced the plasma adrenaline response to alpha LTx but had almost no effect on the noradrenaline response, indicating that alpha LTx-stimulation of sympathetic nerve terminals is direct, whereas catecholamine release from the adrenal medulla is probably mediated by preganglionic release of acetylcholine. In vitro, alpha LTx induced a dose-dependent release of 3H-noradrenaline from rat irides preincubated with this radioactive amine and this effect was not changed by chlorisondamine plus atropine. By contrast, the toxin had no effect on 3H-noradrenaline release from suspensions of cultured rat chromaffin cells. Specific, high affinity binding of 125I-alpha LTx in iris and adrenal medulla homogenates was found to be exceedingly low, suggesting that it might be restricted to nerve terminals. No 125I-alpha LTx binding was seen nor could any effect of the toxin on 14C-5-hydroxytryptamine release be found in rat blood platelet preparations. alpha LTx binding and its amine releasing effect seem, therefore, to be specific for neurons and absent from other cells, even those, like adrenomedullary cells and blood platelets, which share with neurons their origin and/or other important characteristics.

Characterization of α-latrotoxin interaction with rat brain synaptosomes and PC12 cells

α-latrotoxin, a polypeptide neurotoxin purified from the venom of the spider Latrodectus mactans tredecimguttatus , induces a massive release of a variety of neurotransmitters from rat brain synaptosomes and a clonal pheochromocytoma cell line (PC12 cells). In both systems secretion of catecholamines is dose- and calcium-dependent. Efflux of catecholamines is coupled with a substantial release of intracellular ATP. Interaction of α-latrotoxin with PC12 cells is followed by a rapid influx of calcium and sodium ions, the rate being dependent on toxin and calcium concentrations. By reductive methylation it is possible to radioactively label α-latrotoxin without appreciable loss of neurotoxicity. A sensitive binding assay in vitro allows the identification of a limited number of specific binding sites in central nervous system synaptic membranes and PC12 cells, for which tritiated α-latrotoxin displays nanomolar affinity.

A toxin purified from the venom of black widow spider affects the uptake and release of radioactive gamma-amino butyrate and N-epinephrine from rat brain synaptosomes.

A purified neurotoxin from the venom of black widow spider Latrodectus mactans tredecimguttatus, when tested on synaptosomes by means of a perfusion technique, stimulates the release of previously accumulated radioactive transmitters.Black widow spider toxin has also been shown to be a potent inhibitor of the uptake systems for γ‐aminobutyrate and N‐epinephrine. The release from synaptosomes of radioactive N‐epinephrine and γ‐aminobutyrate was rapidly increased by the presence of a low concentration of toxin (∼ 1 nM). The toxin does not appear to cause either non‐specific membrane damage or the release of a non‐transmitter substance like γ‐amino[14C]isobutyric acid.The venom‐induced release of γ‐aminobutyrate was shown not to be dependent on extracellular calcium concentration provided that sodium ions are present. By contrast, that of N‐epinephrine appeared to be Ca‐dependent. The release of γ‐aminobutyrate induced by the toxin is not modified by the presence in the perfusing fluid of tetrodotoxin, dithioerythritol and strontium ions, or by pretreatment of synaptosomes with diaminobutyrate. The mechanism of toxin‐induced release is different from that of a depolarization‐induced release.By correlating previous electrophysiological observations with the results obtained in this biochemical study it may be suggested that the toxin can be used in studies aimed at the identification of components specific to the presynaptic terminal membrane involved in neurotransmitter release.

Preparation and properties of a neurotoxin purified from the venom of black widow spider ( Latrodectus mactans tredecimguttatus)

A neurotoxin of the venom of the spider Latrodectus mactans tredecimguttatus, has been prepared in a homogeneous form and examined by a variety of techniques. The protein has a molecular weight of 130 000 and its toxicity in mice is about 49 000 LD 50 mg pure protein/g body weight. The toxin releases norepinephrine from synaptosomes prepared from rat brain and shows most of the toxic effects of the crude venom preparation.

Effects of chromatographic fractions of black widow spider venom on in vitro biological systems

Extracts of Latrodectus mactans tredecimguttatus venom glands were fractionated by means of gel filtration and ion exchange chromatography. Three protein peaks toxic to mice were assayed in vitro on three different preparations known to be affected by whole venom. The fractions elicited: (a) increased release of acetylcholine from rat brain cortex slices, (b) disappearance of the histochemical reaction due to catecholamines in adrenergic fibres of rat iris, and (c) amplitude decrease of preganglionically evoked action potentials recorded postganglionically from rat superior cervical ganglion, and appearance of asynchronous postganglionic action potentials recorded from unstimulated ganglia. Fractions C and D were active at protein concentrations as low as 2 μg/ml, while fraction B was considerably less potent.

Effects of black widow spider venom on acetylcholine release from rat cerebral cortex slices in vitro

Addition of a whole extract of Latrodectus mactans tredecimguttatus venom glands to rat cerebral cortex slices incubated in an eserinized low-potassium saline solution elicits over the following hour a more than 2-fold increase in the amount of acetylcholine (ACh) released into the medium, and a corresponding decrease in the ACh content of the tissue. The total amount of ACh synthesized in 1 hr is not significantly altered; however, if the tissue is incubated in a high-potassium medium, the resulting enhanced ACh synthesis is markedly reduced. Protein concentrations of venom ranging around 15 μg/ml in the incubation medium are sufficient to elicit a full response. Neurotransmitter depletion of cholinergic nerve terminals may contribute to explain the intoxication symptoms in man and in other vertebrates.

The effect of the venom of the black widow spider Latrodectus mactans tredecimguttatus on the giant neurones of Periplaneta americana

The effects of the venom of the black widow Latrodectus mactans tredecimguttatus have been studied by intracellular recordings from giant neurones at the sixth abdominal ganglion of Periplaneta americana. The venom induces: (i) a progressive decline of the resting potential of the giant neurones; (ii) failure of synaptic transmission through a progressive reduction of the EPSP; (iii) spontaneous repetitive firing of the giant neurones; (iv) alterations of the shape and reduction in size of the propagated action potential. It is suggested that the venom causes a massive release of transmitter which in turn depolarizes the post-synaptic membrane.

Studies on labeled animal poisons—II: Distribution of the venoms of various spiders labeled with Se 75 and P 32 in the guinea pig

P 32- and Se 75-labeled venoms of Latrodectus mactans tredecimguttatus, Steatoda paykulliana, Segestria florentina and Hogna ( Lycosa) tarentula were prepared in vivo. Electrophoretic analysis of Se 75-labeled venom of the tarantula showed rather uniform incorporation of Se 75 in all protein fractions obtained. Radioactivity of labeled venom delivered to the bitten snout of guinea pigs was measured.

Transmission in invertebrate and vertebrate ganglia.

Transmission in invertebrate and vertebrate ganglia.L TaucL TaucPublished Online:01 Jul 1967https://doi.org/10.1152/physrev.1967.47.3.521MoreSectionsPDF (12 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Download PDF FiguresReferencesRelatedInformationCited ByThe effects of acetylcholine, dopamine and noradrenaline on the visceral ganglion of Helix pomatia—I. 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The three species of Latrodectus (Araneae), found in Israel

The three species of widow spiders found in the Negev Desert of Israel, Latrodectus mactans tredecimguttatus, L. pallidus and L. revivensis, seem to differ more in their behaviour and habits than in their morphology. Differences in structure of the genitalia are minor compared with differences in the integument of the abdomen and the structure of the sternum.

Acquired immune response of the house fly, Musca domestica (Linnaeus), to injected venom of the spider Latrodectus mactans tredecimguttatus (Rossi)

Acquired immune response of the house fly, Musca domestica (Linnaeus), to injected venom of the spider Latrodectus mactans tredecimguttatus (Rossi)