Report from the first snake genomics and integrative biology meeting

Animal science

Renal plasticity in response to feeding in the Burmese python, Python molurus bivittatus

Burmese pythons are sit-and-wait predators that are well adapted to go long periods without food, yet subsequently consume and digest single meals that can exceed their body weight. These large feeding events result in a dramatic alkaline tide that is compensated by a hypoventilatory response that normalizes plasma pH; however, little is known regarding how plasma HCO3− is lowered in the days post-feeding. The current study demonstrated that Burmese pythons contain the cellular machinery for renal acid–base compensation and actively remodel the kidney to limit HCO3− reabsorption in the post-feeding period. After being fed a 25% body weight meal plasma total CO2 was elevated by 1.5-fold after 1day, but returned to control concentrations by 4days post-feeding (dpf). Gene expression analysis was used to verify the presence of carbonic anhydrase (CA) II, IV and XIII, Na+ H+ exchanger 3 (NHE3), the Na+ HCO3− co-transporter (NBC) and V-type ATPase. CA IV expression was significantly down-regulated at 3dpf versus fasted controls. This was supported by activity analysis that showed a significant decrease in the amount of GPI-linked CA activity in isolated kidney membranes at 3dpf versus fasted controls. In addition, V-type ATPase activity was significantly up-regulated at 3dpf; no change in gene expression was observed. Both CA II and NHE3 expression was up-regulated at 3dpf, which may be related to post-prandial ion balance. These results suggest that Burmese pythons actively remodel their kidney after feeding, which would in part benefit renal HCO3− clearance

Characterization of carbonic anhydrase XIII in the erythrocytes of the Burmese python, Python molurus bivittatus

Carbonic anhydrase (CA) is one of the most abundant proteins found in vertebrate erythrocytes with the majority of species expressing a low activity CA I and high activity CA II. However, several phylogenetic gaps remain in our understanding of the expansion of cytoplasmic CA in vertebrate erythrocytes. In particular, very little is known about isoforms from reptiles. The current study sought to characterize the erythrocyte isoforms from two squamate species, Python molurus and Nerodia rhombifer, which was combined with information from recent genome projects to address this important phylogenetic gap. Obtained sequences grouped closely with CA XIII in phylogenetic analyses. CA II mRNA transcripts were also found in erythrocytes, but found at less than half the levels of CA XIII. Structural analysis suggested similar biochemical activity as the respective mammalian isoforms, with CA XIII being a low activity isoform. Biochemical characterization verified that the majority of CA activity in the erythrocytes was due to a high activity CA II-like isoform; however, titration with copper supported the presence of two CA pools. The CA II-like pool accounted for 90 % of the total activity. To assess potential disparate roles of these isoforms a feeding stress was used to up-regulate CO2 excretion pathways. Significant up-regulation of CA II and the anion exchanger was observed; CA XIII was strongly down-regulated. While these results do not provide insight into the role of CA XIII in the erythrocytes, they do suggest that the presence of two isoforms is not simply a case of physiological redundancy

Characterization of Bradykinin-related peptides generated in the plasma of six Sarcopterygian species (African lungfish, Amphiuma, Coachwhip, Bullsnake, Gila monster, and Gray's monitor)

Incubation of heat-denatured plasma from six species occupying different evolutionary positions within the Sarcopterygian lineage [the dipnoan, Protopterus annectens (African lungfish); the urodele, Amphiuma tridactylum (three-toed amphiuma); the colubrid snakes, Pituophis melanoleucus sayi (bullsnake) and Masticophis flagellum (coachwhip); and the lizards Heloderma suspectum (Gila monster) and Varanus Grayi (Gray's monitor)] with trypsin generated bradykinin-related peptides that were detected by radioimmunoassay using an antiserum raised against mammalian bradykinin (BK). The peptides were purified by HPLC and their primary structures were established as lungfish [Tyr1,Gly2,Ala7,Pro8]BK, amphiuma [Phe1,Ile2, Leu5]BK, bullsnake and coachwhip [Val1,Thr6]BK, Gila monster [Leu2, Thr6]BK, and Gray's monitor [Thr6]BK. Monitor BK is identical to the peptide generated in turtle and alligator plasma and coachwhip/bullsnake BK shows one amino acid substitution (Ala1 --> Val) compared with the peptide generated in the plasma of the python. The data provide further evidence for the widespread occurrence of a kallikrein-kininogen system in nonmammalian vertebrates but indicate that the primary structure of BK has been poorly conserved during evolution

Phylogeny, Ecology, And Heart Position In Snakes

The cardiovascular system of all animals is affected by gravitational pressure gradients, the intensity of which varies according to organismic features, behavior, and habitat occupied. A previous nonphylogenetic analysis of heart position in snakes-which often assume vertical postures-found the heart located 15%-25% of total body length from the head in terrestrial and arboreal species but 25%-45% in aquatic species. It was hypothesized that a more anterior heart in arboreal species served to reduce the hydrostatic blood pressure when these animals adopt vertical postures during climbing, whereas an anterior heart position would not be needed in aquatic habitats, where the effects of gravity are less pronounced. We analyzed a new data set of 155 species from five major families of Alethinophidia (one of the two major branches of snakes, the other being blind snakes, Scolecophidia) using both conventional and phylogenetically based statistical methods. General linear models regressing log10 snout-heart position on log10 snout-vent length (SVL), as well as dummy variables coding for habitat and/or clade, were compared using likelihood ratio tests and the Akaike Information Criterion. Heart distance to the tip of the snout scaled isometrically with SVL. In all instances, phylogenetic models that incorporated transformation of the branch lengths under an Ornstein-Uhlenbeck model of evolution (to mimic stabilizing selection) better fit the data as compared with their nonphylogenetic counterparts. The bestfit model predicting snake heart position included aspects of both habitat and clade and indicated that arboreal snakes in our study tend to have hearts placed more posteriorly, opposite the trend identified in previous studies. Phylogenetic signal in relative heart position was apparent both within and among clades. 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