Expanding the invertebrate medicine toolbox: evaluation of opisthosoma tonometry as a novel diagnostic tool for arachnids.

OBJECTIVE: The objective of this study was to analyze the use of rebound tonometry, as a noninvasive diagnostic tool, in arachnids. METHODS: 5 juvenile (yearling) female and 1 juvenile male curlyhair tarantulas (Tliltocatl albopilosus, previously Brachypelma albopilosum) were used to track estimated ventral opisthosoma pressures over a 9-month period of time. Younger, growing animals were selected as they are more likely than adults to go through multiple molts throughout the 9 months of the study length. An iCare TONOVET TV01 rebound tonometer was used to measure the estimated ventral opisthosoma pressures of the spiders. Measurements were obtained from the ventral opisthosoma, 1 of the thinnest areas of exocuticle throughout the body. Readings were obtained once per week from the ventral opisthosoma for the first 2 months, then once every 2 weeks for 1 month, then back to once per week for the remainder of the study. Additional measurements were obtained following each ecdysis, after a 2% body weight sampling of hemolymph, and at the end of the study to evaluate readings in response to induction of general anesthesia with 5% isoflurane gas and oxygen flow at 2 L/min for 10 minutes. RESULTS: The average of all estimated ventral opisthosoma pressure when spiders were not in molt was 26.19 mm Hg (SD, 3.54), with a statistically significant decrease postmolt to an average of 15.31 mm Hg (SD, 3.81), followed by a gradual increase back to premolt pressures over a 3-week period with an average of 22 days (SD, 1.93). Estimated ventral opisthosoma pressures decreased post hemolymph removal. There is not sufficient evidence that estimated ventral opisthosoma pressures changed over time following the anesthesia. CLINICAL RELEVANCE: This study demonstrates that tonometry can be used to generally assess the estimated ventral opisthosoma pressure, which could correlate with where a spider is in an ecdysis cycle.

Envenomation by the Indian ornamental tarantula (Poecilotheria regalis): A case report on treatment with Latrodectus mactans antivenom.

Poecilotheria spiders are considered theraphosids of underestimated clinical importance, with bites from these species inducing symptoms such as severe pain and intense muscle cramps. However, there is no specific treatment for the envenomation caused by these species, which, while native to India and Sri Lanka, are widely distributed worldwide. The present study reports the case of a 31-year-old man bitten by a Poecilotheria regalis specimen. The patient's clinical presentation was similar to Latrodectus envenomation, and patient was treated with an L. mactans antivenom. Most of patient's symptoms improved (fasciculations, pain, erythema, and local swelling), except muscle cramps. A toxicological study conducted on mice did not show that L. mactans antivenom has a neutralizing effect on the toxicity of P. regalis. The present report discusses the envenoming process of Poecilotheria species and the possible neutralizing effect exerted by L. mactans antivenom.

Perioral Paresthesia, Generalize Muscle Cramps, and Rhabdomyolysis After a Socotra Island Blue Baboon Tarantula (Monocentropus Balfouri) Bite.

BACKGROUND: Literature on systemic envenomation caused by tarantula bites, particularly from the Theraphosidae family, is relatively scarce. This case report provides a formal description of the first known instance of systemic envenomation caused by the Socotra Island Blue Baboon Tarantula (Monocentropus balfouri). CASE REPORT: In this case, a 23-year-old employee of an exotic pet shop suffered from perioral paresthesia, generalized muscle cramps, and rhabdomyolysis because of a Monocentropus balfouri bite. His symptoms were successfully relieved with oral benzodiazepines. EMERGENCY PHYSICIAN BE AWARE OF THIS?: This case highlights the potential for serious complications resulting from the bite of Monocentropus balfouri, a species gaining popularity among global exotic pet collectors.

Innovative tarantula hair-inspired washing machine filters for enhanced microfiber capture.

Aquatic environments are being polluted by microplastics primarily originating from the washing of synthetic textiles. Microfibers (MF), which are microplastics in synthetic fibers, are consistently introduced into the environment with each domestic laundry cycle. To address this issue, we developed a specialized MF capture "barbed filter" (BF) by transforming PET monofilaments of different diameters (0.4, 0.6, and 0.8 mm) into structures that closely resemble the characteristics of tarantula urticating hairs. BFs feature sharp barbs that effectively capture and retain microfibers of diverse lengths, surpassing the performance of conventional control filters. The BFs had a retention efficiency of 88-91 %, while the CFs had an efficiency of 79-86 %. Our findings revealed that the barbed filter significantly outperformed the conventional control filter in capturing microfibers due to its smaller pore size, shorter pore distance, and unique filter shape. This design not only enhances the surface area and friction, facilitating microfibril strong entrapment but also minimizes the probability of microfibril passage through the filter. This research offers a promising solution for reducing microfibril release from laundry and textile industrial wastewater. The implementation of BFs in real life has the potential to significantly reduce microplastic pollution and promote a cleaner and more sustainable environment.

All about being old and shooting hairs: clade age and urticating hair explain the patterns of diversification in tarantulas.

The extreme asymmetry of species richness distribution across the tree of life has always intrigued evolutionary biologists. Two competing explanations have been proposed to explain this pattern-the clade age hypothesis and diversification rate variation. While these two scenarios may not be mutually exclusive, to what extent time and diversification rates interact to explain species richness patterns remains understudied. Here, we investigate the relative influence of these two scenarios using tarantulas (Family: Theraphosidae) as a model. Tarantulas represent a speciose group of spiders found worldwide but exceptionally diverse in South America. These spiders show two distinct patterns of microhabitat use (ground-dwelling or arboreal) and defense strategies (presence or absence of urticating hairs). Using various trait-independent and dependent diversification models, we test the clade age hypothesis, the role of microhabitat, antipredator defense strategy, and geography in influencing diversification rates. Our results suggest that clade age is the primary predictor of species richness distribution across the tarantula subfamilies. However, the presence of urticating hair probably disrupted this pattern in some clades by increasing the net diversification rates, not by increasing the speciation rate but by reducing the extinction rate.

A Case Report on Generalised Muscle Spasms After Ornamental Tarantula Bites That Responded Well to Intravenous Calcium Gluconate Treatment.

Here, we present a case of a 55-year-old male, who was admitted with a spider bite, which caused swelling of the hand and painful muscle spasms along with palpitations. The patient made a complete recovery after the administration of intravenous calcium gluconate, followed by oral calcium supplements. Although no specific treatment exists in Sri Lanka for spider bites, calcium supplements can be beneficial for Sri Lankan ornamental tarantula (Poecilotheria fasciata) bites.

Increasing knowledge of Cymbiapophysa Gabriel & Sherwood, 2020 (Araneae, Theraphosidae): general distribution, key to species, and three new species from Ecuador.

Three new species of Cymbiapophysa Gabriel & Sherwood, 2020 are described from south, central, and north-western Ecuador, showing the wide range of distribution that this genus has in Ecuador and its biogeographical provinces. These three new species are easily differentiated from other congeners based on keel morphology of the male palpal bulb. Supplementary information about the locality of C.magna Sherwood, Gabriel, Brescovit & Lucas, 2021 is provided, alongside additional data on morphology and some commentaries about the general distribution and biogeography of Cymbiapophysa. Additionally, a taxonomic key for males of Cymbiapophysa species is presented, based on the palpal bulb morphology.

A Case Report of Venezuelan Suntiger Tarantula (Psalmopoeus Irminia) Envenomation and Review of Tarantula Exposures.

BACKGROUND: Tarantula envenomations are encountered infrequently but may increase with increased exotic animal ownership. This case report presents the first documented toxicity from a Venezuelan suntiger tarantula (VST), Psalmopoeus irminia, and provides a general framework for approaching patients with tarantula exposures. CASE REPORT: A 35-year-old man presented to an emergency department 4 h after experiencing a bite from his pet VST. He developed erythema, pain, and edema to the bite site on the left thenar eminence that extended proximally. Within 4 h, he developed abdominal pain, nausea, vomiting, throat itching, and tightness. The patient had a blood pressure of 131/105 mm Hg, heart rate of 102 beats/min, 36.6°C, respiratory rate of 20 breaths/min, and SpO2 of 94%. Laboratory evaluations were within normal limits (other than chronically elevated but improved transaminases). The patient received 0.5 mg epinephrine intramuscularly, 50 mg diphenhydramine IV, 20 mg famotidine IV, 0.4 mg ondansetron IV, and 1 L of normal saline for a suspected anaphylactic reaction. Shortly after epinephrine administration, his gastrointestinal and upper airway symptoms resolved. All symptoms resolved within 1 week. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Little is known about VST toxicity. Therefore, providers should rely on a general framework for approaching patients with tarantula exposures. Morbidity from tarantula exposures is mediated by mechanical injury, venom effects, and hypersensitivity reactions. Typical clinical findings include local pain, pruritis, edema, erythema, and burning. Muscle cramping, ophthalmia nodosa, and hypersensitivity reactions may occur. Treatment is primarily supportive and includes decontamination, cool compresses, analgesia, treatment of anaphylaxis, and ophthalmology evaluation if ocular exposure.

Indian Ornamental Tarantula (Poecilotheria regalis) Venom Affects Myoblast Function and Causes Skeletal Muscle Damage.

Envenomation by the Indian ornamental tarantula (Poecilotheria regalis) is medically relevant to humans, both in its native India and worldwide, where they are kept as pets. Muscle-related symptoms such as cramps and pain are commonly reported in humans following envenomation by this species. There is no specific treatment, including antivenom, for its envenomation. Moreover, the scientific knowledge of the impact of this venom on skeletal muscle function is highly limited. Therefore, we carried out this study to better understand the myotoxic properties of Poecilotheria regalis venom by determining its effects in cultured myoblasts and in the tibialis anterior muscle in mice. While there was no effect found on undifferentiated myoblasts, the venom affected differentiated multinucleated myotubes resulting in the reduction of fusion and atrophy of myotubes. Similarly, intramuscular administration of this venom in the tibialis anterior muscle in mice resulted in extensive muscle damage on day 5. However, by day 10, the regeneration was evident, and the regeneration process continued until day 20. Nevertheless, some tissue abnormalities including reduced dystrophin expression and microthrombi presence were observed on day 20. Overall, this study demonstrates the ability of this venom to induce significant muscle damage and affect its regeneration in the early stages. These data provide novel mechanistic insights into this venom-induced muscle damage and guide future studies to isolate and characterise individual toxic component(s) that induce muscle damage and their significance in developing better therapeutics.

The effects of tarantula cubensis extract on renal ischemia reperfusion injury in rats.

BACKGROUND: Renal ischemia-reperfusion (IR) related acute kidney injury (AKI) is an important health problem and has not yet been fully treated. Tarantula cubensis extract (TCE) is a homeopathic drug that has antiinflammatory and antioxidant effects. This study aimed to investigate the effects of TCE on renal ischemia-reperfusion injury in rats. METHODS: This study was carried out on 48 Spraque-Dawley male rats, which were divided into six groups. The first, second, and third groups were control, sham, and IR groups, respectively. Group four received IR and 0.2 mL of 96% ethanol. Group five and six received ischemia and reperfusion and TCE 0.01 and 0.1 mg per rat (which correspond to approximately 0.04 mg/kg, and 0.4 mg/kg), respectively. Tumor necrosis factor alpha (TNF-α), interleukin-1beta (IL-1ß), total antioxidant status (TAS), and total oxidant status (TOS) levels in renal tissue were measured by enzyme-linked immunosorbent assay (ELISA). Oxidative stress index (OSI) was obtained by proportioning TAS and TOS. Superoxide dismutase (SOD), myeloperoxidase (MPO) activities, and malondialdehyde (MDA) levels were determined by manual spectrophotometric methods. The histopathological changes were evaluated via hematoxylineosin and immunohistochemical staining. RESULTS: In IR group, renal tissue TNF-α and IL-1ß levels were significantly higher than control group (p < 0.0001 for both), and low(p < 0.0001 for both) and high dose (p < 0.0001 for both) TCE administration decreased these markers. Low and high doses of TCE decreased OSI values compared with IR group (p = 0.04 and p = 0.001 respectively). Although TCE decreased MDA levels, it was not statistically significant. MPO levels significantly decreased. In addition, TCE has been found to prevent hemorrhage, cast formation, and dilatation caused by IR in renal tissues stained with hematoxylin-eosin. And also, the most intense nuclear factor kappa B (NFκB) and caspase-3 immunopositivity found in IR group was decreased in both of the TCE groups. DISCUSSION: Although TCE showed a protective effect by inhibiting inflammation against IR damage in renal tissues, there was no clear effect on oxidative stress. Larger and more detailed studies are needed to clarify the issue.

Expression of posterior Hox genes and opisthosomal appendage development in a mygalomorph spider.

Spiders represent an evolutionary successful group of chelicerate arthropods. The body of spiders is subdivided into two regions (tagmata). The anterior tagma, the prosoma, bears the head appendages and four pairs of walking legs. The segments of the posterior tagma, the opisthosoma, either lost their appendages during the course of evolution or their appendages were substantially modified to fulfill new tasks such as reproduction, gas exchange, and silk production. Previous work has shown that the homeotic Hox genes are involved in shaping the posterior appendages of spiders. In this paper, we investigate the expression of the posterior Hox genes in a tarantula that possesses some key differences of posterior appendages compared to true spiders, such as the lack of the anterior pair of spinnerets and a second set of book lungs instead of trachea. Based on the observed differences in posterior Hox gene expression in true spiders and tarantulas, we argue that subtle changes in the Hox gene expression of the Hox genes abdA and AbdB are possibly responsible for at least some of the morphological differences seen in true spiders versus tarantulas.

Four new species of mygalomorph spiders (Araneae, Halonoproctidae and Theraphosidae) from the Colombian Pacific region (Bahía Solano, Chocó).

The Colombian Pacific coast is an amazing natural region, immersed in one of the most unknown biodiversity hotspots in the world. An expedition carried out in the north of this area, at the Jardín Botánico del Pacífico (JBP) in Bahía Solano, Chocó, focused on studying the diversity of the mygalomorph spider fauna, allowed us to discover four new species included in the families Halonoproctidae and Theraphosidae. The trapdoor species Ummidiasolanasp. nov., and the theraphosids species Euthycaeluscunampiasp. nov. (Schismatothelinae), Melloinapacificasp. nov. (Glabropelmatinae), and Neischnocolusmecanasp. nov. (Theraphosinae) are illustrated, diagnosed, and described in detail. Photographs of somatic features and copulatory organs and a distribution map are provided. Morphological, taxonomical, and biogeographical aspects are discussed for each species. All these taxonomic novelties represent the first records of these genera for the region, expanding the range of geographic distribution of each of them. This work constitutes the first effort focused on characterizing the community of Mygalomorphae species in the Chocó Biogeographic Region.

Vasodilator activity of Poecilotheria ornata venom involves activation of the NO/cGMP pathway and inhibition of calcium influx to vascular smooth muscle cells.

Tarantula venoms may be a natural source of new vasodilator components useful in pharmacological research. Moreover, biological function data of the venoms are important to enhance the knowledge about the biodiversity and evolution of these species. The present study aims to describe the vasodilatory activity induced by the venom of Poecilotheria ornata on isolated rat aortic rings. This venom induced a vasodilator activity that was significantly reduced after incubation with L-NAME or ODQ. Measurements of nitrite concentrations on rat aorta homogenates showed that the venom significantly increased the basal levels. Moreover, the venom attenuates the contraction induced by calcium. These results suggest that P. ornata venom contains a mixture of vasodilator components that act through the activation of the nitric oxide/cGMP pathway, as well as, through an endothelium-independent mechanism that involves the calcium influx into vascular smooth muscle cells.

Using the Integrative Approach to Update a Gap of One Century: Redescription and New Distribution Records of the South American Tarantulas Grammostola pulchra (Araneae: Mygalomorphae: Theraphosidae).

Taxonomic researchers have used multiple sources of evidence to support species hypotheses and delimitations. Grammostola Simon (Mygalomorphae: Theraphosidae) comprises 20 valid species endemic to South America, six occurring in Brazil. The classical morphological approach based mainly on genitalia may be misleading in recognizing species in this genus. Thus, we used morphology, geographical distribution, genetic distance, and phylogeny to support the redescription of Grammostola pulchra from southern Brazil, a species described a century ago. We also diagnosed and illustrated the species. Males have a developed apical keel at the apex of the embolus; for the first time, this type of structure has been reported in a species of Grammostola. The molecular analyses using the partial sequence of Cytochrome c oxidase subunit I showed 7% of genetic distance (p-distance) between G. pulchra and Grammostola anthracina. Distance and tree-based methods (ASAP and bPTP, respectively) assigned G. pulchra as a valid species. The gene-tree under Bayesian and Maximum-Likelihood recovered a similar topology, placing G. pulchra as closely related to Grammostola burzaquensis and G. anthracina. Morphological characters which could be important in the taxonomy of the genus are further discussed.

Retrospective analysis of histologic lesions in captive arachnids.

Invertebrates, including arachnids, are a common taxon in zoological collections. Invertebrate medicine and pathology are emerging subspecialties, but there is limited reference material or published resources describing histologic lesions in arachnids. Histopathology of 26 captive arachnids (20 spiders and 6 scorpions) from institutional collections was reviewed. Most animals were found dead with limited clinical signs. Tissues evaluated included body wall (cuticle and epidermis), skeletal muscle, book lungs, digestive tract (pharynx, esophagus, sucking stomach, midgut tube, midgut diverticula, and stercoral pocket), central and peripheral nervous system, heart, hemolymph vessels and sinuses, Malpighian tubules, coxal glands, and gonads. Inflammation was frequent (24/26, 92%), and seen in multiple organs (18/24, 75%) with the midgut diverticulum most commonly affected (14/24, 58%) followed by the book lungs (13/24 arachnids, 54%), and body wall (8/24 arachnids, 33%). Inflammation comprised hemocyte accumulation, hemocytic coagula, melanization, and nodulation. Infectious agents, including bacteria (11/26, 42%), fungi (10/26, 38%), and parasites (2/26, 8%), were seen within inflammatory aggregates. Coinfection with multiple infectious agents was common (6/24, 25%). No etiologic agent was identified in 7/24 (29%) cases with inflammatory lesions. Lesions suggestive of decreased nutritional status or increased metabolic rate included midgut diverticula atrophy in 11/26 (42%) animals and skeletal muscle atrophy in 6/26 (23%) animals. Atrophic lesions were seen in combination with infection (8/11, 73%), pregnancy (2/11, 18%), male sex (2/11, 18%), or without other lesions (1/11, 9%). Other suspected contributors to death included dysecdysis-associated trauma (2/26, 8%) and uterine intussusception (1/26, 4%). No animals had neoplasia.

Wessel's tiger ornamental tarantula bite envenomation: A case report and venom analysis.

Tarantulas are commonly kept as pets and bites from some species can cause severe symptoms. Here we describe a case of a patient with transient atrial fibrillation (afib) and painful muscle cramps requiring hospitalization for pain management after being bitten by a Poecilotheria tigrinawesseli (Wessel's Tiger Ornamental) spider. He was discharged with a cardiac event monitor and outpatient cardiology follow-up. The event monitor documented transient afib which decreased in frequency then resolved halfway through the three-week monitoring period. In conclusion, tarantula envenomation is usually mild with local pain and edema most reported. However, bites by some species, such as P. tigrinawesseli may have local and more systemic, long-lasting effects.

Effect of intravesical tarantula cubensis extract (Theranekron) on inflammation in an interstitial cystitis rat model.

OBJECTIVES: To reveal the histopathological and immunological outcomes of intravesical treatment with tarantula cubensis extract (TCE) in a rat model of interstitial cystitis. METHODS: A total of 30 female Wistar albino rats were divided into three groups: group 1 (control group), group 2 (disease group), and group 3 (treatment group). The rat model of interstitial cystitis was created by biweekly intraperitoneal administration of cyclophosphamide (CYP). In group 3, TCE (a venom extracted from a brown spider known as tarantula cubensis) was administered intravesically after the model had been created. Urothelial degeneration, necrosis, ulcer, bleeding, edema, inflammation and mast cell count, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), myeloperoxidase (MPO), and hydroxyproline parameters were evaluated. Statistical analysis was performed using one-way analysis of variance, chi-square tests, and Kruskal-Wallis tests. RESULTS: All parameters were found to be lower in the rats in group 1 than in the other groups, and IL-6 and MPO values were found to be higher in group 2 (p < .001). The mean TNF-alpha value was highest in group 2 (p = .078). No difference was found between all groups regarding ulcer (p = .087). Urothelial degeneration, necrosis, edema, inflammation, hemorrhage and fibroblast proliferations, and hydroxyproline values were higher in group 3 (p < .001). CONCLUSIONS: Intravesical TCE instillation produces an anti-inflammatory effect by reducing the levels of inflammatory parameters such as IL-6, TNF-alpha, and MPO in bladder tissue. It also accelerates tissue healing by increasing hydroxyproline and fibroblast proliferation.

The effect of Tarantula cubensis D6 on zone of stasis in a rat burn model.

AIM: Burn injuries are one of the most devastating injuries. Saving the zone of stasis decreases burn size, morbidity, and mortality. Tarantula cubensis (TC) increases epithelization of wounds, and increases wound contraction. In this study, we investigated the effects of TC on the stasis zone. MATERIALS AND METHODS: 36 Wistar albino female rats were divided into 3 groups. Using brass comb burn model, at days 0 and 3, physiological serum solution in group 2, TC injection in group 3and no injection in group 1 was applied. Thermal and normal images were taken on day 10 and rats were sacrificed for histopathological examination. RESULTS: We found a statistically significant difference between the 1st and 3rd group, 2nd and 3rd group regarding viable wound areas (p < 0.05), temperature difference (body-stasis zone) and mean body temperature (p < 0.05). We found no statistically significant difference between groups regarding neovascularization, inflammatory density, and vital cutaneous appendages. We found a statistically significant difference in epithelial layer thickness between the1st and 3rd groups, 2nd and 3rd groups(p < 0.05). CONCLUSION: TC increases stasis zone viability macroscopically, increases epithelial layer thickness histopathologically, thus it promotes wound healing in burn wounds. This increase in stasis zone viability was also established with thermal imaging.

Improving the Annotation of the Venom Gland Transcriptome of Pamphobeteus verdolaga, Prospecting Novel Bioactive Peptides.

Spider venoms constitute a trove of novel peptides with biotechnological interest. Paucity of next-generation-sequencing (NGS) data generation has led to a description of less than 1% of these peptides. Increasing evidence supports the underestimation of the assembled genes a single transcriptome assembler can predict. Here, the transcriptome of the venom gland of the spider Pamphobeteus verdolaga was re-assembled, using three free access algorithms, Trinity, SOAPdenovo-Trans, and SPAdes, to obtain a more complete annotation. Assembler's performance was evaluated by contig number, N50, read representation on the assembly, and BUSCO's terms retrieval against the arthropod dataset. Out of all the assembled sequences with all software, 39.26% were common between the three assemblers, and 27.88% were uniquely assembled by Trinity, while 27.65% were uniquely assembled by SPAdes. The non-redundant merging of all three assemblies' output permitted the annotation of 9232 sequences, which was 23% more when compared to each software and 28% more when compared to the previous P. verdolaga annotation; moreover, the description of 65 novel theraphotoxins was possible. In the generation of data for non-model organisms, as well as in the search for novel peptides with biotechnological interest, it is highly recommended to employ at least two different transcriptome assemblers.

The Tarantula Toxin ω-Avsp1a Specifically Inhibits Human CaV3.1 and CaV3.3 via the Extracellular S3-S4 Loop of the Domain 1 Voltage-Sensor.

Inhibition of T-type calcium channels (CaV3) prevents development of diseases related to cardiovascular and nerve systems. Further, knockout animal studies have revealed that some diseases are mediated by specific subtypes of CaV3. However, subtype-specific CaV3 inhibitors for therapeutic purposes or for studying the physiological roles of CaV3 subtypes are missing. To bridge this gap, we employed our spider venom library and uncovered that Avicularia spec. ("Amazonas Purple", Peru) tarantula venom inhibited specific T-type CaV channel subtypes. By using chromatographic and mass-spectrometric techniques, we isolated and sequenced the active toxin ω-Avsp1a, a C-terminally amidated 36 residue peptide with a molecular weight of 4224.91 Da, which comprised the major peak in the venom. Both native (4.1 µM) and synthetic ω-Avsp1a (10 µM) inhibited 90% of CaV3.1 and CaV3.3, but only 25% of CaV3.2 currents. In order to investigate the toxin binding site, we generated a range of chimeric channels from the less sensitive CaV3.2 and more sensitive CaV3.3. Our results suggest that domain-1 of CaV3.3 is important for the inhibitory effect of ω-Avsp1a on T-type calcium channels. Further studies revealed that a leucine of T-type calcium channels is crucial for the inhibitory effect of ω-Avsp1a.