The mechanisms that control the sizes of a body and its many parts remain among the great puzzles in developmental biology. Why do animals grow to a species-specific body size, and how is the relative growth of their body parts controlled to so they grow to the right size, and in the correct proportion with body size, giving an animal its species-characteristic shape? Control of size must involve mechanisms that somehow assess some aspect of size and are upstream of mechanisms that regulate growth. These mechanisms are now beginning to be understood in the insects, in particular in Manduca sexta and Drosophila melanogaster. The control of size requires control of the rate of growth and control of the cessation of growth. Growth is controlled by genetic and environmental factors. Insulin and ecdysone, their receptors and intracellular signaling pathways are the principal genetic regulators of growth. The secretion of these growth hormones, in turn, is controlled by complex interactions of other endocrine and molecular mechanisms, by environmental factors such as nutrition and by the physiological mechanisms that sense body size. Although the general mechanisms of growth regulation appear to be widely shared, the mechanisms that regulate final size can be quite diverse.

Body size profoundly affects many aspects of animal biology, including metamorphosis, allometry, size-dependent alternative pathways of gene expression, and the social and ecological roles of individuals. However, regulation of body size is one of the fundamental unsolved problems in developmental biology. The control of body size requires a mechanism that assesses size and stops growth within a characteristic range of sizes. Under normal growth conditions in Manduca sexta, the endocrine cascade that causes the brain to initiate metamorphosis starts when the larva reaches a critical weight. Metamorphosis is initiated by a size-sensing mechanism, but the nature of this mechanism has remained elusive. Here we show that this size-sensing mechanism depends on the limited ability of a fixed tracheal system to sustain the oxygen supply to a growing individual. As body mass increases, the demand for oxygen also increases, but the fixed tracheal system does not allow a corresponding increase in oxygen supply. We show that interinstar molting has the same size-related oxygendependent mechanism of regulation as metamorphosis. We show that low oxygen tension induces molting at smaller body size, consistent with the hypothesis that under normal growth conditions, body size is regulated by a mechanism that senses oxygen limitation. We also found that under poor growth conditions, larvae may never attain the critical weight but eventually molt regardless. We show that under these conditions, larvae do not use the critical weight mechanism, but instead use a size-independent mechanism that is independent of the brain. M ost species of animals experience determinate growth and stop growing within a characteristic range of body size (1-4). This raises the question of how body size is sensed such that it is regulated within this range. How body size is sensed remains one of the fundamental unsolved problems in developmental biology.In insect larvae, the signal to stop growing and initiate a molt is the secretion of the steroid hormone ecdysone. Larval-larval molts are caused by pulses of ecdysone, and in the last larval stage of the tobacco hornworm, Manduca sexta, ecdysone causes the larva to stop feeding and initiate metamorphosis (5). The timing of ecdysone secretion is determined by the critical weight (6-9), a size threshold at which the endocrine events that eventually result in the cessation of feeding, entry into the wandering stage, and metamorphosis are initiated. The discovery of the critical weight demonstrated that initiation of the metamorphic molt depends not on instar duration or growth rate, but rather on size.The time course of these endocrine events is independent of further nutrition and growth. Therefore, the critical weight can be operationally assessed as the weight at which nutrition is no longer necessary for a normal time course to the cessation of feeding, entry into the wandering stage, and the pupal molt (5, 6, 9). The weight that we measure is a proxy for a physiological variable that is correlated w...

The developmental mechanisms that control body size and the relative sizes of body parts are today best understood in insects. Size is controlled by the mechanisms that cause growth to stop when a size characteristic of the species has been achieved. This requires the mechanisms to assess size and respond by stopping the process that controls growth. Growth is controlled by two hormones, insulin and ecdysone, that act synergistically by controlling cell growth and cell division. Ecdysone has two distinct functions: At low concentration it controls growth, and at high levels it causes molting and tissue differentiation. Growth is stopped by the pulse of ecdysone that initiates the metamorphic molt. Body size is sensed by either stretch receptors or oxygen restriction, depending on the species, which stimulate the high level of ecdysone secretion that induces a molt. Wing growth occurs mostly after the body has stopped growing. Wing size is adjusted to body size by variation in both the duration and level of ecdysone secretion.

Body size determination requires a mechanism for sensing size and a mechanism for linking size information to the termination of growth. Although the hormonal mechanisms that terminate growth are well elucidated, the mechanisms by which a body senses its own size are only partially understood; most of this understanding has come from the study of the mechanisms that control insect moulting and metamorphosis. We first review and discuss advances in our understanding of the physiological mechanisms by which insect larvae sense their size. Second, we present new findings on how larvae in which the size-sensing mechanism has been disrupted eventually terminate growth (in a size-independent manner). We synthesize recent insights into the genetic and molecular mechanisms of ecdysteroid regulation in Drosophila melanogaster with developmental physiology findings in Manduca sexta, paving the way for an integrated understanding of the mechanisms of body size regulation.

Holometabolous insects undergo dramatic morphological and physiological changes during ontogeny. In particular, the larvae of many holometabolous insects are specialized to feed in soil, water or dung, inside plant structures, or inside other organisms as parasites where they may commonly experience hypoxia or anoxia. In contrast, holometabolous adults usually are winged and live with access to air. Here, we show that larval Drosophila melanogaster experience severe hypoxia in their normal laboratory environments; third instar larvae feed by tunneling into a medium without usable oxygen. Larvae move strongly in anoxia for many minutes, while adults (like most other adult insects) are quickly paralyzed. Adults survive anoxia nearly an order of magnitude longer than larvae (LT50: 8.3 versus 1 h). Plausibly, the paralysis of adults is a programmed response to reduce ATP need and enhance survival. In support of that hypothesis, larvae produce lactate at 3× greater rates than adults in anoxia. However, when immobile in anoxia, larvae and adults are similarly able to decrease their metabolic rate, to about 3% of normoxic conditions. These data suggest that Drosophila larvae and adults have been differentially selected for behavioral and metabolic responses to anoxia, with larvae exhibiting vigorous escape behavior likely enabling release from viscous anoxic media to predictably normoxic air, while the paralysis behavior of adults maximizes their chances of surviving flooding events of unpredictable duration. Developmental remodeling of behavioral and metabolic strategies to hypoxia/anoxia is a previously unrecognized major attribute of holometabolism.

SUMMARYRearing oxygen level is known to affect final body size in a variety of insects, but the physiological mechanisms by which oxygen affects size are incompletely understood. In Manduca sexta and Drosophila melanogaster, the larval size at which metamorphosis is initiated largely determines adult size, and metamorphosis is initiated when larvae attain a critical mass. We hypothesized that oxygen effects on final size might be mediated by oxygen effects on the critical weight and the ecdysone titers, which regulate growth rate and the timing of developmental transitions. Our results showed that oxygen affected critical weight, the basal ecdysone titers and the timing of the ecdysone peak, providing clear evidence that oxygen affected growth rate and developmental rate. Hypoxic third instar larvae (10% oxygen) exhibited a reduced critical weight, slower growth rate, delayed pupariation, elevated baseline ecdysone levels and a delayed ecdysone peak that occurred at a lower larval mass. Hyperoxic larvae exhibited increased basal ecdysone levels, but no change in critical weight compared with normoxic larvae and no significant change in timing of pupariation. Previous studies have shown that nutrition is crucial for regulating growth rate and the timing of developmental transitions. Here we show that oxygen level is one of multiple cues that together regulate adult size and the timing and dynamics of growth, developmental rate and ecdysone signaling. Supplementary material available online at

Ichthyostega and Acanthostega are the earliest tetrapods known from multiple near-complete skeletons, with Acanthostega generally considered the more primitive. New material indicates differing ontogenetic trajectories for their forelimbs: In Ichthyostega, the pattern of muscle attachment processes on small humeri (upper arm bones) resembles that in "fish" members of the tetrapod stem group such as Tiktaalik, whereas large humeri approach (but fail to attain) the tetrapod crown-group condition; in Acanthostega, both small and large humeri exhibit the crown-group pattern. We infer that Ichthyostega underwent greater locomotory terrestrialization during ontogeny. The newly recognized primitive characteristics also suggest that Ichthyostega could be phylogenetically more basal than Acanthostega.

Insect tracheal-respiratory systems achieve high fluxes and great dynamic range with low energy requirements and could be important models for bioengineers interested in developing microfluidic systems. Recent advances suggest that insect cardiorespiratory systems have functional valves that permit compartmentalization with segment-specific pressures and flows and that system anatomy allows regional flows. Convection dominates over diffusion as a transport mechanism in the major tracheae, but Reynolds numbers suggest viscous effects remain important.