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PubMed2006

Snake venom glutaminyl cyclase.

Glutaminyl cyclase (QC) catalyzes N-terminal glutamine cyclization of many endocrine peptides and is typically abundant in brain tissue. As three-finger toxins in the venoms of colubrid snakes Boiga dendrophila and Boiga irregularis contain N-terminal pyroglutamate, we searched for QC in venom glands of both snakes. Here we report cDNA sequences of QC from brain and venom gland tissues of Boiga species. We propose that QC expressed in snake venom gland tissue plays a role in the N-terminal pyroglutamate formation of several snake venom toxins, indirectly contributing to venom potency.

Amino Acid Sequence↗
PubMed2006

Systemic pathological alterations caused by Philodryas patagoniensis colubrid snake venom in rats.

Very little is known about the systemic effects caused by Philodryas patagoniensis colubrid snake venom. In this work, this venom was tested for its ability to induce histopathological changes in rats after its intramuscular, subcutaneous or intravenous administration, by light microscopic examination of some organs (cerebellum, cerebrum, lung, liver, kidney and heart). Four rats were used for each dose of 0.23, 0.45 and 0.90 mg of venom in 0.3 ml of phosphate-buffered saline solution (pH 7.4). Aliquots of blood were withdrawn at different time intervals for enzymatic determination of alanine aminotransferase, aspartate aminotransferase and creatine kinase levels. After 2h the animals were killed by an overdose of anesthetic, and samples of kidney, heart, liver, lung, cerebrum and cerebellum were taken to microscopic examination (hematoxylin and eosin stain). Histologically, no abnormality was observed in heart tissue, in none of the administration routes of the venom used. However, histological observations showed multifocal hemorrhage in cerebellum, cerebrum and lung sections, severe peritubular capillary congestion in kidney sections and hydropic degeneration in liver sections, when venom was administrated intravenously. The subcutaneous route showed similar results to the previous one, with the exception of cerebellar hemorrhage. Intramuscularly, neither cerebral nor cerebellar hemorrhage was observed. Plasma alanine aminotransferase and aspartate aminotransferase increased levels were demonstrated, mainly when venom was administered intravenously or subcutaneously. Our results suggest that P. patagoniensis venom induces moderate histopathological changes in vital organs of rats. These changes are initiated at early stages of the envenomation and may be associated with a behavioral or functional abnormality of those organs during envenoming.

Alanine Transaminase↗
PubMed2005

A serotonin receptor antagonist, but not melatonin, modulates hormonal responses to capture stress in two populations of garter snakes (Thamnophis sirtalis parietalis and Thamnophis sirtalis concinnus).

Hormonal and behavioral responses to a stressor depend on many factors, including the influence of other hormones. We examined the role of melatonin in modulating hormonal responses to capture stress in two populations of male garter snakes, Thamnophis sirtalis. Studies of red-sided (T. sirtalis parietalis) and red-spotted (T. sirtalis concinnus) garter snakes were conducted in the field with free-living snakes. Populations of red-sided garter snakes in south-central Manitoba, Canada undergo a period of winter dormancy for approximately 8 months each year followed by an attenuated mating season (4-5 weeks) in early spring. In contrast, the mid-latitude red-spotted garter snake in western Oregon, USA has an extended breeding season and can be active during 10-12 months of the year given appropriate environmental conditions. We chose to study these two populations of garter snakes to investigate possible variation in melatonin function among snakes with different suites of environmental adaptations. To better address these questions, we also examined the effects of 5-hydroxytryptophan (a precursor of melatonin synthesis) and ketanserin (a serotonergic type 2A receptor antagonist) on hormonal responses to capture stress. We observed a trend of increased corticosterone and decreased androgen concentrations in northern-latitude red-sided garter snakes (T. sirtalis parietalis) subjected to 4 h of capture stress during the spring. However, these differences were not statistically significant. During the fall, red-sided garter snakes showed no change in corticosterone or androgen concentrations in response to the capture stress treatments. We speculate that northern-latitude red-sided garter snakes suppress hormonal responses to capture stress during preparation for winter dormancy. Treatment with melatonin, 5-hydroxytryptophan, or ketanserin did not significantly influence corticosterone or androgen concentrations of northern-latitude red-sided garter snakes during the spring or fall. Mid-latitude red-spotted garter snakes (T. sirtalis concinnus) from Oregon showed a statistically significant increase in corticosterone concentrations in response to 4 h of capture stress; treatment with melatonin, 5-hydroxytryptophan, or ketanserin prior to capture stress had no significant influence on plasma corticosterone concentrations. Androgen concentrations of mid-latitude red-spotted garter snakes in response to capture stress were significantly lower than those of non-stressed control snakes. Neither melatonin nor 5-hydroxytryptophan influenced the change in androgen concentrations during capture stress. However, androgen concentrations of snakes treated with ketanserin prior to 4 h of capture stress did not differ significantly from those of non-stressed control snakes. These studies suggest that melatonin does not modulate hormonal responses to capture stress in this ectothermic model. Our results also suggest that a serotonin-regulated system may play a role in modulating the activity of the hypothalamic-pituitary-gonadal axis during physiological stress responses.

5-Hydroxytryptophan↗
PubMed2006

Corticosterone and the transition from courtship behavior to dispersal in male red-sided garter snakes (Thamnophis sirtalis parietalis).

Seasonal modulation of baseline glucocorticoid concentrations as well as the sensitivity of the hypothalamic-pituitary-adrenal (HPA) axis plays an important role in supporting critical life-history events such as seasonal reproduction and migration. Despite numerous studies on adrenocortical modulation, little is known about the exact timing of this seasonal modulation with respect to critical life-history stages. We tested the hypothesis that seasonal modulation of the HPA axis during the spring mating season in male red-sided garter snakes (Thamnophis sirtalis parietalis) is temporally linked to the mechanisms regulating dispersal. We compared hormonal responses to capture stress in courting male red-sided garter snakes collected from the den site and den perimeter to those of dispersing snakes collected 0.6 km from the den. We also investigated possible changes in steroid hormones during the spring mating season. These studies support previous findings that plasma androgen and corticosterone concentrations significantly decline over the mating season. Our results demonstrate that males 0.6 km into a 15-20 km route to the feeding grounds have lower baseline corticosterone concentrations than male snakes actively courting at the den. Dispersing males also exhibit a typical stress response marked by a significant increase in corticosterone while actively courting males do not. Capture stress did not significantly influence androgen concentrations of either courting or dispersing male red-sided garter snakes. There were no significant differences in body composition indices among male snakes collected from the den, den perimeter, or 0.6 km away from the den. However, we did observe a significant negative correlation between baseline corticosterone levels and body composition indices. These data suggest that breeding is a distinct stage accompanied by specific physiological parameters that differ from those during dispersal to the feeding grounds. Our results indicate that declining baseline corticosterone concentrations may play a role in the behavioral switch between actively courting and dispersing (i.e., feeding) in the late spring.

Analysis of Variance↗
PubMed2004

Effects of melatonin on the behavioral and hormonal responses of red-sided garter snakes (Thamnophis sirtalis parietalis) to exogenous corticosterone.

We investigated possible interactions between melatonin and corticosterone in modulating the reproductive behavior of male red-sided garter snakes (Thamnophis sirtalis parietalis) following spring emergence. We also examined whether melatonin's modulatory actions could be explained by its potential properties as a serotonin receptor antagonist. Exogenous corticosterone significantly reduced courtship behavior of male snakes in a dose-dependent manner. Melatonin also significantly reduced courtship behavior of male garter snakes. Pretreatment with melatonin before administering corticosterone treatments further suppressed courtship behavior of red-sided garter snakes. These results indicate additive inhibitory effects of melatonin and corticosterone in modulating reproductive behavior. Snakes receiving ketanserin, a serotonergic type 2A receptor antagonist, followed by corticosterone also showed reduced courtship behavior; this serotonin receptor antagonist followed by treatment with vehicle did not significantly influence courtship behavior of male snakes. Neither melatonin nor corticosterone treatments significantly influenced testosterone + 5-alpha-dihydrotestosterone concentrations of male garter snakes, supporting a direct effect of melatonin and corticosterone on courtship behavior that is independent of any effect on androgen concentrations. We propose that a serotonin system is involved in the modulation of male courtship behavior by melatonin and corticosterone. In addition, our data support the hypothesis that melatonin may function as a serotonin receptor antagonist. Further research is necessary to discern whether the actions of melatonin and corticosterone are converging on the same pathway or if their effects on different pathways are having additive inhibitory effects on courtship behavior.

Analysis of Variance↗
PubMed2004

Relict snakes of North America and their relationships within Caenophidia, using likelihood-based Bayesian methods on mitochondrial sequences.

This paper focuses on the phylogenetic relationships of eight North American caenophidian snake species (Carphophis amoena, Contia tenuis, Diadophis punctatus, Farancia abacura, Farancia erytrogramma, Heterodon nasicus, Heterodon platyrhinos, and Heterodon simus) whose phylogenetic relationships remain controversial. Past studies have referred to these "relict" North American snakes either as colubrid, or as Neotropical dipsadids and/or xenodontids. Based on mitochondrial DNA ribosomal gene sequences and a likelihood-based Bayesian analysis, our study suggests that these North American snakes are not monophyletic and are nested within a group (Dipsadoidea) that contains the Dipsadidae, Xenodontidae, and Natricidae. In addition, we use the relationships proposed here to highlight putative examples of parallel evolution of hemipenial morphology among snake clades.

Animals↗
PubMed2006

How and when did Old World ratsnakes disperse into the New World?

To examine Holarctic snake dispersal, we inferred a phylogenetic tree from four mtDNA genes and one scnDNA gene for most species of the Old World (OW) and New World (NW) colubrid group known as ratsnakes. Ancestral area distributions are estimated for various clades using divergence-vicariance analysis and maximum likelihood on trees produced using Bayesian inference. Dates of divergence for the same clades are estimated using penalized likelihood with statistically crosschecked calibration references obtained from the Miocene fossil record. With ancestral areas and associated dates estimated, various hypotheses concerning the age and environment associated with the origin of ratsnakes and the dispersal of NW taxa from OW ancestors were tested. Results suggest that the ratsnakes originated in tropical Asia in the late Eocene and subsequently dispersed to the Western and Eastern Palearctic by the early Oligocene. These analyses also suggest that the monophyletic NW ratsnakes (the Lampropeltini) diverged from OW ratsnakes and dispersed through Beringia in the late Oligocene/early Miocene when this land bridge was mostly composed of deciduous and coniferous forests.

Animals↗
PubMed2006

Post-glacial recolonization of the Great Lakes region by the common gartersnake (Thamnophis sirtalis) inferred from mtDNA sequences.

Pleistocene events played an important role in the differentiation of North American vertebrate populations. Michigan, in particular, and the Great Lakes region, in general, were greatly influenced by the last glaciation. While several hypotheses regarding the recolonization of this region have been advanced, none have been strongly supported. We generated 148 complete ND2 mitochondrial DNA (mtDNA) sequences from common gartersnake (Thamnophis sirtalis) populations throughout the Great Lakes region to evaluate phylogeographic patterns and population structure and to determine whether the distribution of haplotypic variants is related to the post-Pleistocene retreat of the Wisconsinan glacier. The common gartersnake was utilized, as it is believed to have been one of the primary vertebrate invaders of the Great Lakes region following the most recent period of glacial retreat and because it has been a model species for a variety of evolutionary, ecological, behavioral, and physiological studies. Several genetically distinct evolutionary lineages were supported by both genealogical and molecular population genetic analyses, although to different degrees. The geographic distribution of the majority of these lineages is interpreted as reflecting post-glacial recolonization dynamics during the late Pleistocene. These findings generally support previous hypotheses of range expansion in this region.

Animals↗
PubMed2006

Postprandial responses in the African rhombig egg eater (Dasypeltis scabra).

The African rhombic egg eater (Dasypeltis scabra) is a colubrid snake feeding exclusively on bird eggs. Frequency of feeding is governed by the seasonal availability of bird eggs; i.e., long fasting intervals change with relatively short periods when plenty of food is available. Intermittent feeding snakes show a remarkable postprandial increase of metabolic rate and digestive organ size. The postprandial increase in metabolic rate (specific dynamic action, SDA) in snakes is affected by meal size, temperature, and meal composition. A major portion of SDA in snakes is allocated to gastric function and the breakdown of the meal. We hypothesize that SDA in egg eaters is lower than in other snake species, because egg eaters feed on "liquid" food that does not require enzymatic breakdown in the stomach. We also hypothesized that other components of the postprandial response of egg eaters (e.g., size changes of the intestine and the liver) do not differ from other snakes. The standard metabolic rate and metabolic response to feeding were measured using closed-chamber respirometry. Size changes of small intestine and liver were measured using high-resolution transcutaneous ultrasonography. Standard metabolic rates of fasting egg eaters were in the same range of mass specific values as known from other snakes. Within 24h after feeding, oxygen consumption doubled and peaked at 2 days after feeding. At the same time, the size of the small intestine and the cross-sectional diameter of the liver increased. Within 2 days after feeding, the size of the mucosal epithelium doubled its thickness. Liver size increased significantly within 24h reaching maximum size 2-4 days after feeding. The size of both organs returned to fasting values within 7-10 days after feeding. The postprandial response of African rhombic egg eaters shows the same pattern and dynamics as known from other snake species. However, the factorial increase of metabolic rate during SDA is the lowest reported for any snake. A comparison with literature data supports the idea that SDA is mainly determined by gastric function and that it is low in egg eaters because they do not have to break down solid meals in the stomach as other snake species do.

Animals↗
PubMed2006

Alpha- and beta-keratins of the snake epidermis.

Snake scales contain specialized hard keratins (beta-keratins) and alpha- or cyto-keratins in their epidermis. The number, isoelectric point, and the evolution of these proteins in snakes and their similarity with those of other vertebrates are not known. In the present study, alpha- and beta-keratins of snake molts and of the whole epidermis have been studied by using two-dimensional electrophoresis and immunocytochemistry. Specific keratins in snake epidermis have been identified by using antibodies that recognize acidic and basic cytokeratins and avian or lizard scale beta-keratin. Alpha keratins of 40-70 kDa and isoelectric point (pI) at 4.5-7.0 are present in molts. The study suggests that cytokeratins in snakes are acidic or neutral, in contrast to mammals and birds where basic keratins are also present. Beta keratins of 10-15 kDa and a pI of 6.5-8.5 are found in molts. Some beta-keratins appear as basic proteins (pI 8.2) comparable to those present in the epidermis of other reptiles. Some basic "beta-keratins" associate with cytokeratins as matrix proteins and replace cytokeratins forming the corneous material of the mature beta-layer of snake scales, as in other reptiles. The study also suggests that more forms of beta-keratins (more than three different types) are present in the epidermis of snakes.

Animals↗
PubMed2002

Molecular cloning and characterization of protein phosphatase 2C of vomeronasal sensory epithelium of garter snakes.

The earthworm-derived chemoattractant ES20 interacts with its G-protein-coupled receptors on the plasma membrane of vomeronasal (VN) sensory neurons of garter snakes, resulting in an increase in inositol trisphosphate [J. Biol. Chem. 269 (1994) 16867] and a rapid phosphorylation of the membrane-bound proteins, p42/44 [Biochim. Biophys. Acta 1450 (1999) 320]. The phosphorylation of p42/44 proteins are countervailingly regulated by a protein kinase and an okadaic acid-insensitive but fluoride-sensitive protein phosphatase (PPase) [J. Liu et al. (loc. cit.)]. The phosphorylation of p42/44 induced by ES20 appears to play a role in the regulation of signal transduction pathways by modulating the GTPase activity [J. Liu et al. (loc. cit.)]. A 564-bp fragment of cDNA was obtained from VN RNA of garter snakes by reverse transcription polymerase chain reaction with degenerate primers. The 564-bp fragment was amplified, cloned, and sequenced. Northern blot analysis revealed that both the VN organ (VNO) and brain contained the gene of PPase 2C. A full-length complementary 4119-bp DNA containing an open reading frame of 1146bp that encodes a protein of 382 amino acids with a molecular mass of 49,123Da was obtained from the VN cDNA library of garter snakes. The deduced amino acid sequence showed 88% amino acid identity to bovine protein phosphatase 2C alpha and 87% identity to human and rat PP2C alpha and to Mg(2+)-dependent protein phosphatase 1A of rat and rabbit. In situ hybridization revealed that the mRNA of VN protein phosphatase 2C is expressed in the vomeronasal sensory epithelium. This is the first report of the identification of a type 2C serine/threonine protein phosphatase in the VN system.

Amino Acid Sequence↗
PubMed2002

Immunohistochemical identification of components of the chemoattractant signal transduction pathway in vomeronasal bipolar neurons of garter snakes.

The chemosignal transduction pathway in the vomeronasal sensory epithelium of garter snakes involves activation of G-protein-coupled receptors and subsequent generation of second messengers leading to production of an electrical signal. Calcium imaging experiments demonstrate that ligand binding to the receptor leads to an increase in intracellular calcium and that the phosphatidylinositol-turnover pathway plays a major role in this Ca(2+) increase. Here, we demonstrate, using immunohistochemistry, that IP(3) receptors are largely distributed in dendritic regions of the epithelium, ryanodine receptors are confined to the somata region, and Na(+)/Ca(2+) exchanger protein is expressed throughout the vomeronasal (VN) sensory epithelium.

Animals↗
PubMed1997

Development of the plerocercoid I of Ophiotaenia europaea in reptiles.

The existence of a two-host life-cycle in ophiotaeniid proteocephalideans was tested experimentally using Ophiotaenia europaea as a model. Three species of reptiles, Natrix natrix, Natrix tessellata and Lacerta viridis, were fed with experimentally infected copepods containing a large number of infective plerocercoids I. A few plerocercoids, most of which were dead, corresponding morphologically to the plerocercoid II developmental stage of O. europaea, were found encysted in the intestinal wall of N. natrix (8 days p.i.), N. tessellata (5 and 150 days p.i.) and L. viridis (40 days p.i.), while no plerocercoids or adult worms were recovered from their intestines. The results indicate that the infective plerocercoid I of O. europaea cannot undergo further development when ingested directly by the final host (a reptile), and that environmental temperature stimuli cannot initiate a reverse plerocercoid migration to the gut followed by strobilization.

Animals↗
PubMed2002

First structural evidence of a specific inhibition of phospholipase A2 by alpha-tocopherol (vitamin E) and its implications in inflammation: crystal structure of the complex formed between phospholipase A2 and alpha-tocopherol at 1.8 A resolution.

This is the first structural evidence of alpha-tocopherol (alpha-TP) as a possible candidate against inflammation, as it inhibits phospholipase A2 specifically and effectively. The crystal structure of the complex formed between Vipera russelli phospholipase A2 and alpha-tocopherol has been determined and refined to a resolution of 1.8 A. The structure contains two molecules, A and B, of phospholipase A2 in the asymmetric unit, together with one alpha-tocopherol molecule, which is bound specifically to one of them. The phospholipase A2 molecules interact extensively with each other in the crystalline state. The two molecules were found in a stable association in the solution state as well, thus indicating their inherent tendency to remain together as a structural unit, leading to significant functional implications. In the crystal structure, the most important difference between the conformations of two molecules as a result of their association pertains to the orientation of Trp31. It may be noted that Trp31 is located at the mouth of the hydrophobic channel that forms the binding domain of the enzyme. The values of torsion angles (phi, psi, chi(1) and chi(2)) for both the backbone as well as for the side-chain of Trp31 in molecules A and B are -94 degrees, -30 degrees, -66 degrees, 116 degrees and -128 degrees, 170 degrees, -63 degrees, -81 degrees, respectively. The conformation of Trp31 in molecule A is suitable for binding, while that in B hinders the passage of the ligand to the binding site. Consequently, alpha-tocopherol is able to bind to molecule A only, while the binding site of molecule B contains three water molecules. In the complex, the aromatic moiety of alpha-tocopherol is placed in the large space at the active site of the enzyme, while the long hydrophobic channel in the enzyme is filled by hydrocarbon chain of alpha-tocopherol. The critical interactions between the enzyme and alpha-tocopherol are generated between the hydroxyl group of the six-membered ring of alpha-tocopherol and His48 N(delta1) and Asp49 O(delta1) as characteristic hydrogen bonds. The remaining part of alpha-tocopherol interacts extensively with the residues of the hydrophobic channel of the enzyme, giving rise to a number of hydrophobic interactions, resulting in the formation of a stable complex.

Animals↗
PubMed1997

Gonadotropin antagonist modulates courtship behavior in male red-sided garter snakes, Thamnophis sirtalis parietalis.

Behavioral studies were used to investigate the central effects of chicken-I GnRH, chicken-II GnRH, and D-Phe2,6,Pro3-GnRH, a GnRH antagonist, on the courtship behavior of male red-sided garter snakes, Thamnophis sirtalis parietalis. Intracerebroventricular (i.c.v.) injections of chicken-I or chicken-II GnRH had no effect on time spent courting or latency to court when experimental males were exposed to unmated females, or when experimental males were exposed to the female sex attractiveness pheromone. I.c.v. injections of D-Phe2,6,Pro3-GnRH caused a significant decrease in latency to court when experimental males were exposed to unmated females. When males injected with D-Phe2,6,Pro3-GnRH were exposed to the female sex attractiveness pheromone, it caused a significant increase in time spent courting compared to that in saline-injected controls. D-Phe2,6,Pro3-GnRH was not able to initiate courtship behavior during the nonbreeding season, indicating that courtship behavior is dependent on the interaction of multiple components. This study does demonstrate that a hormone or neuropeptide can modulate sexual behavior in garter snakes.

Animal Communication↗
PubMed2001

Airway ammonia negates the normal ventilatory response to airway CO(2) in garter snakes.

Earlier studies from this lab showed that unidirectionally ventilated snakes, maintained on high airway [CO(2)], breathe slowly with a large tidal volume. If airway [CO(2)] is abruptly reduced during inspiration, inspiratory duration (TI) and tidal volume increase. On the other hand, in an animal normally receiving fresh air (no CO(2) in the inspired air) on each inspiration, if fresh air is withheld for one breath, TI and tidal volume decrease. To test the effect of producing an alkaline environment in the presence of CO(2) during these two maneuvers, six unidirectionally ventilated snakes weighing 32+/-16 g were maintained on 4% CO(2) in air flowing at 300 ml/min even during inspiration. NH(3) (4%) was introduced into the lung during one inspiration. During that breath, TI increased significantly, as if CO(2) had been removed from the lung. In another study, six different snakes weighing 21+/-6 g received fresh air at each inspiration. At the end of inspiration [CO(2)] was again raised to 4%. However, for one breath, [CO(2)] was maintained high during inspiration and airway NH(3) (3--4%) was simultaneously introduced into the airstream. During this breath, TI was expected to decrease, owing to the elevated [CO(2)]; however, there was no significant difference in the TI of the test breath compared with the previous breath, indicating that ammonia interfered with the expected CO(2) response. This study suggests that pH (probably intracellular) rather than P(CO(2)) produces this ventilatory reflex.

Administration, Inhalation↗
PubMed2000

Mass spectrophotometric evidence for P-III/P-IV metalloproteinases in the venom of the Boomslang (Dispholidus typus).

The Boomslang, Dispholidus typus, is a mid- to rear-fanged arboreal colubrid widely distributed throughout much of the African continent. Envenoming by this species is rare although deaths have been recorded. Typical symptoms associated with envenoming include diffuse intravascular coagulation (DIC) caused by fibrinogen consumption and consequent incoagulable blood together with haemorrhage into tissues such as muscle and brain; together, these procoagulant and haemorrhagic effects of the venom result in a very poor prognosis in patients who receive a large dose of venom and who are not treated with antivenom. Renal failure may also result from acute tubular necrosis resulting from pigment nephropathy. Little is known about the toxic components present in the venom; however, proteolytic activity has been reported although the proteinases involved have not been identified. In this study we provide LC/MS/MS (liquid chromatography/mass spectrometry/mass spectrometry) data supporting the presence of class P-III/P-IV snake venom metalloproteinases (SVMPs) in Boomslang venom. Using a polyclonal antibody raised against the P-III haemorrhagic toxin (Jararhagin) obtained from the venom of the Brazilian pit viper, Bothrops jararaca, we identified by western blot a 65 kDa protein from Boomslang venom which cross-reacted with the jararhagin antibody. A corresponding band from SDS-PAGE was subjected to tryptic digestion followed by LC/MS/MS sequence analysis of the digestion mixture. A variety of peptide sequences were identified in the digest, one of which was clearly homologous with a highly conserved region of the disintegrin-like domains of P-III/P-IV SVMPs. These data provide the first structural evidence for the presence of SVMPs in Boomslang venom; it is possible that SVMPs may also be present in the venoms of other colubrids, which cause similar symptoms in envenomed humans. In other snake venoms, most notably those of the Viperinae and Crotalinae subfamilies, many of the coagulopathic and haemorrhagic syndromes associated with systemic and local envenoming are attributed to SVMPs. The identification of a P-III/P-IV SVMP sequence in D. typus venom suggests that many of the pathological signs resulting from envenoming by this species may also be due to the presence of SVMPs in the venom. It is hoped that these results may accelerate research into colubrid venoms and may provide new insights into novel and more efficacious treatments for colubrid envenoming.

Amino Acid Sequence↗
PubMed2000

Characterization of venom (Duvernoy's secretion) from twelve species of colubrid snakes and partial sequence of four venom proteins.

R.E. Hill and S.P. Mackessy. Characterization of venom (Duvernoy's secretion) from twelve species of colubrid snakes and partial sequence of four venom proteins. Toxicon XX, xx-yy, 2000. - Venomous colubrids, which include more than 700 snake species worldwide, represent a vast potential source of novel biological compounds. The present study characterized venom (Duvernoy's gland secretion) collected from twelve species of opisthoglyphous (rear-fanged) colubrid snakes, an extremely diverse assemblage of non-venomous to highly venomous snakes. Most venoms displayed proteolytic activity (casein), though activity levels varied considerably. Low phosphodiesterase activity was detected in several venoms (Amphiesma stolata, Diadophis punctatus, Heterodon nasicus kennerlyi, H. n. nasicus and Thamnophis elegans vagrans), and acetylcholinesterase was found in Boiga irregularis saliva and venom, but no venoms displayed hyaluronidase, thrombin-like or kallikrein-like activities. High phospholipase A(2) (PLA(2)) activity was found in Trimorphodon biscutatus lambda venom, and moderate levels were detected in Boiga dendrophila and D. p. regalis venoms as well as B. dendrophila and H. n. nasicus salivas. Non-reducing SDS-PAGE revealed 7-20 protein bands (3.5 to over 200 kD, depending on species) for all venoms analyzed, and electrophoretic profiles of venoms were typically quite distinct from saliva profiles. Components from A. stolata, Hydrodynastes gigas, Tantilla nigriceps and T. e. vagrans venoms showed protease activity when run on gelatin zymogram gels. N-terminal protein sequences for three 26 kD venom components of three species (H. gigas, H. torquata, T. biscutatus) and one 3.5 kD component (T. nigriceps) were also obtained, and the 3.5 kD peptide showed apparent sequence homology with human vascular endothelial growth factor; these data represent the first sequences of colubrid venom components. Protease, phosphodiesterase and PLA(2) activities are also common to elapid and viperid snake venoms, but it is apparent that numerous other (as yet undescribed) components make up the majority of colubrid venom proteins. The complex nature of venoms produced by most species surveyed, and the high levels of protease or phospholipase A(2) activity of some venoms, suggest that many colubrids could become an important source of human health concern as encounters with these snakes increase.

Acetylcholinesterase↗