Ultrastructure and Morphometric Analysis of Interstitial Cells of Cajal in the Gastric Wall of the Bullfrog (Rana catesbeiana)

Interstitial cells of Cajal (ICC) play a vital role in the gastrointestinal motility. However, information on ICC in lower vertebrates is rare. Here, ICC and ICC-like features of the gastric wall in the bullfrog (Rana catesbeiana) were observed by light microscopy and transmission electron microscopy. The lengths and distances of the ICC/ICC-like features were measured by morphometric analysis. The gastric wall contained mucosa, submucosa, tunica muscularis, and serosa. The gastric glands contained mucous cells and oxynticopeptic cells. The ICC with 1–3 processes were located among smooth muscle cells (SMC) of the tunica muscularis. Moreover, the ICC-like features were observed among oxynticopeptic cells of the mucosa. The processes of ICC established direct contacts with SMC. Also, the gap junctions were observed between the processes of ICC and nerve fiber bundles in the tunica muscularis. The multivesicular bodies, including shedding exosomes, were frequently observed between ICC and SMC. In addition, ICC-like features and their processes were observed in close proximity to oxynticopeptic cells and blood vessels. Our findings illustrated that ICC are present in the gastric tunica muscularis, and ICC-like features were in the mucosal lamina propria of the gastric wall of R. catesbeiana. These histological evidences supported the notion that ICC are implicated in gastric motility.

Identification and characterization of amphibian SLC26A5 using RNA-Seq

Background Prestin (SLC26A5) is responsible for acute sensitivity and frequency selectivity in the vertebrate auditory system. Limited knowledge of prestin is from experiments using site-directed mutagenesis or domain-swapping techniques after the amino acid residues were identified by comparing the sequence of prestin to those of its paralogs and orthologs. Frog prestin is the only representative in amphibian lineage and the studies of it were quite rare with only one species identified. Results Here we report a new coding sequence of SLC26A5 for a frog species, Rana catesbeiana (the American bullfrog). In our study, the SLC26A5 gene of Rana has been mapped, sequenced and cloned successively using RNA-Seq. We measured the nonlinear capacitance (NLC) of prestin both in the hair cells of Rana’s inner ear and HEK293T cells transfected with this new coding gene. HEK293T cells expressing Rana prestin showed electrophysiological features similar to that of hair cells from its inner ear. Comparative studies of zebrafish, chick, Rana and an ancient frog species showed that chick and zebrafish prestin lacked NLC. Ancient frog’s prestin was functionally different from Rana. Conclusions We mapped and sequenced the SLC26A5 of the Rana catesbeiana from its inner ear cDNA using RNA-Seq. The Rana SLC26A5 cDNA was 2292 bp long, encoding a polypeptide of 763 amino acid residues, with 40% identity to mammals. This new coding gene could encode a functionally active protein conferring NLC to both frog HCs and the mammalian cell line. While comparing to its orthologs, the amphibian prestin has been evolutionarily changing its function and becomes more advanced than avian and teleost prestin.

Characterizations of Gelatin from the Skin of American Bullfrog (Rana catesbeiana) as Affected by Extraction Temperature

We investigated the effect of extraction temperature on the gel properties of gelatin from the skin of the American bullfrog (Rana catesbeiana) and the mechanisms. The textural and rheological properties of bullfrog gelatin extracted at 45 °C (G45), 55 °C (G55), and 65 °C (G65) were measured. The molecular weight distributions, microstructures, and amino acid compositions of the bullfrog gelatins were also determined. G45, G55, and G65 had gel strengths of 272.1, 225.6, and 205.8 g and hardness values of 28.1, 24.0, and 22.5 N, respectively. The gelling temperatures ranged from 19.3 to 23.9 °C, and the melting temperatures ranged from 28.9 to 31.5 °C. All the results were compared with those of commercial porcine gelatin. We propose that the higher gel strength of G45 with a higher band intensity of α2-chains compared with G55 and G65 was more likely to form ordered and strong cross-links. The gelatin extracted at a lower temperature (G45) had a finer gel structure, suggesting that it would be more difficult to disrupt by applied force. Gelatin extracted at a lower temperature demonstrated better properties with α2-chains and a fine gel structure. These results provide basic information on the extraction of American bullfrog skin gelatin for industrial applications.

Historical museum collections and contemporary population studies implicate roads and introduced predatory bullfrogs in the decline of western pond turtles

The western pond turtle (WPT), recently separated into two paripatrically distributed species (Emys pallida and Emys marmorata), is experiencing significant reductions in its range and population size. In addition to habitat loss, two potential causes of decline are female-biased road mortality and high juvenile mortality from non-native predatory bullfrogs (Rana catesbeiana). However, quantitative analyses of these threats have never been conducted for either species of WPT. We used a combination of historical museum samples and published and unpublished field studies shared with us through personal communications with WPT field researchers (B. Shaffer, P. Scott, R. Fisher, C. Brown, R. Dagit, L. Patterson, T. Engstrom, 2019, personal communications) to quantify the effect of roads and bullfrogs on WPT populations along the west coast of the United States. Both species of WPT shift toward increasingly male biased museum collections over the last century, a trend consistent with increasing, female-biased road mortality. Recent WPT population studies revealed that road density and proximity were significantly associated with increasingly male-biased sex ratios, further suggesting female-biased road mortality. The mean body size of museum collections of E. marmorata, but not E. pallida, has increased over the last 100 years, consistent with reduced recruitment and aging populations that could be driven by invasive predators. Contemporary WPT population sites that co-occur with bullfrogs had significantly greater average body sizes than population sites without bullfrogs, suggesting strong bullfrog predation on small WPT hatchlings and juveniles. Overall, our findings indicate that both species of WPT face demographic challenges which would have been difficult to document without the use of both historical data from natural history collections and contemporary demographic field data. Although correlational, our analyses suggest that female-biased road mortality and predation on small turtles by non-native bullfrogs are occurring, and that conservation strategies reducing both may be important for WPT recovery.