Budgett's frog (Lepidobatrachus laevis): A new amphibian embryo for developmental biology.

The large size and rapid development of amphibian embryos has facilitated ground-breaking discoveries in developmental biology. Here, we describe the embryogenesis of the Budgett's frog (Lepidobatrachus laevis), an unusual species with eggs that are over twice the diameter of laboratory Xenopus, and embryos that can tolerate higher temperatures to develop into a tadpole four times more rapidly. In addition to detailing their early development, we demonstrate that, like Xenopus, these embryos are amenable to explant culture assays and can express exogenous transcripts in a tissue-specific manner. Moreover, the steep developmental trajectory and large scale of Lepidobatrachus make it exceptionally well-suited for morphogenesis research. For example, the developing organs of the Budgett's frog are massive compared to those of most model species, and are composed of larger individual cells, thereby affording increased subcellular resolution of early vertebrate organogenesis. Furthermore, we found that complete limb regeneration, which typically requires months to achieve in most vertebrate models, occurs in a matter of days in the Budgett's tadpole, which substantially accelerates the pace of experimentation. Thus, the unusual combination of the greater size and speed of the Budgett's frog model provides inimitable advantages for developmental studies-and a novel inroad to address the mechanisms of spatiotemporal scaling during evolution.

Temporal pattern cues in vibrational risk assessment by embryos of the red-eyed treefrog, Agalychnis callidryas.

The embryos of red-eyed treefrogs, Agalychnis callidryas, use vibrations transmitted through their arboreal egg clutch to cue escape hatching behavior when attacked by egg-eating snakes. Hatching early increases the risk of predation in the water, so embryos should avoid it unless they are in danger. We exposed egg clutches to intermittent vibrations with different combinations of vibration duration and spacing to examine the role of simple temporal pattern cues in the escape hatching response. Stimuli were bursts of synthetic white noise from 0 to 100 Hz, including the range of frequencies with substantial energy in snake attacks, and had approximately rectangular amplitude envelopes. Embryos hatched in response to a small range of temporal patterns and not in response to many others, rather than hatching to most vibrations except for certain patterns perceived as safe. Neither cycle length nor duty cycle predicted hatching response, except at extreme values where no hatching occurred; the highest energy stimuli elicited little or no hatching. Both vibration duration and inter-vibration interval strongly affected the hatching response. The highest levels of hatching were to durations of 0.5 s combined with intervals of 1.5-2.5 s, and hatching decreased gradually with increasing difference of either duration or interval from these most effective stimuli. Vibration duration and interval appear to function as two necessary elements of a composite cue, rather than as redundant cues. This increases response specificity and reduces the range of stimuli that elicit hatching, likely reducing the chance of hatching unnecessarily in a benign disturbance. Vibration-cued hatching in A. callidryas embryos offers an opportunity to experimentally assess the behavioral decision rules underlying an effective and costly anti-predator defense.

Effects of temperature on energy cost and timing of embryonic and larval development of the terrestrially breeding moss frog, Bryobatrachus nimbus.

The Australian moss frog, Bryobatrachus nimbus, oviposits four to 16 large eggs in terrestrial nests constructed in moss or lichen in subalpine regions of southern Tasmania. Nidicolous larvae overwinter beneath snow, reaching metamorphosis without feeding after 395 d, the longest development time known for an endotrophic anuran. However, a few clutches develop more quickly and metamorphose before winter. This study examines the effect of temperature on development time and energy expenditure by measuring temperatures and developmental stages in field nests as well as rates of oxygen consumption (Vo2), developmental stage, body mass, and energy content in the laboratory at three relevant temperatures (5 degrees, 10 degrees, 15 degrees C). Eggs and larvae reared at 5 degrees C differentiated very slowly, and their development time far exceeded those in natural nests, but development times at 10 degrees and 15 degrees C averaged 277 and 149 d, respectively, and were shorter than field incubation times. Generally, respiration rates of aquatic hatchlings were low in comparison with other species but increased with larval age and jumped about 25% higher near metamorphosis when larvae were able to air breathe. The mean energy density was 26.0 J mg(-1) for the dry ova and 20.6 J mg(-1) for a dry gut-free froglet, and total production efficiency was 61.5%. We developed a model based on the relationships between incubation temperature and V&d2;o2 to estimate the respiratory cost of development to metamorphosis, the first such study for an amphibian. The cost was 177 J at 15 degrees C, 199 J at 10 degrees C, and at least 249 J at 5 degrees C, and we predicted that continual development at 5 degrees C would lead to premature yolk depletion because it equalled the 249 J contained in fresh ova. Continuously logged field-nest temperatures and interpolation of laboratory data provided estimates of development rates, Vo2, and respiratory energy costs in field nests. Development to metamorphosis required between 185 and 234 J when larvae overwintered, but completion of metamorphosis before winter saved 123 J. However, the advantage of emergence in warmer months, when conditions are suitable for feeding and growth, may offset the greater energy cost of overwintering.