The developmental and physiological basis of body size evolution in an insect

The evolution of body size is a dominant feature of animal evolution. However, little is known about how the underlying developmental mechanisms that determine size change as body size evolves. Here we report on a case of body size evolution in the tobacco hornworm Manduca sexta that occurred over a period of nearly 30 years. We take advantage of an extensive series of physiological studies performed in the early 1970s that established the parameters that regulate body size in this species and compare their values with those of modern individuals that are descendants of the same colony. We show that three of the five processes that determine adult body size changed during this period, while two remained constant. Changes in these three developmental processes completely account for the observed evolutionary change in body size.     

Homologies in the colour patterns of the genus Charaxes (Lepidoptera: Nymphalidae)

The phylogenetically and morphologically diverse patterns of Charaxes can be reduced to a simple set of pattern elements which can be homologized throughout the genus. At least five types of correspondence (homologies) exist among pattern elements: those between (1) species, (2) forewing and hindwing, (3) dorsal and ventral wing surface, (4) serial wing-cells, and (5) individual pattern elements within a single wing-cell. Differences in Charaxes colour patterns result from the distortion, elaboration, enlargement, reduction or loss of individual pattern elements. Further variation is often the result of dislocation of pattern elements from their serial homologues in neighbouring wing-cells, and fusion of individual pattern elements to create larger areas of colour. The type of analysis presented in this paper should be broadly applicable within the Lepidoptera and may prove useful in studying the systematics of colour patterns and the evolution of the developmental system that gives rise to them.     

A mathematical model of the methionine cycle

Building on the work of Martinov et al. (2000), a mathematical model is developed for the methionine cycle. A large amount of information is available about the enzymes that catalyse individual reaction steps in the cycle, from methionine to S-adenosylmethionine to S-adenosylhomocysteine to homocysteine, and the removal of mass from the cycle by the conversion of homocysteine to cystathionine. Nevertheless, the behavior of the cycle is very complicated since many substrates alter the activities of the enzymes in the reactions that produce them, and some can also alter the activities of other enzymes in the cycle. The model consists of four differential equations, based on known reaction kinetics, that can be solved to give the time course of the concentrations of the four main substrates in the cycle under various circumstances. We show that the behavior of the model in response to genetic abnormalities and dietary deficiencies is similar to the changes seen in a wide variety of experimental studies. We conduct computational "experiments" that give understanding of the regulatory behavior of the methionine cycle under normal conditions and the behavior in the presence of genetic variation and dietary deficiencies.     

Control of growth and differentiation of the wing imaginal disk of Precis coenia (Lepidoptera: Nymphalidae)

During the last larval instar, the wing imaginal disks of Precis coenia grow continuously. The rate of disk growth is not disk-autonomous but closely matches the rate of somatic growth of the larva, so that the size of the disks is a function of the size of the body, irrespective of the growth rate of the larva. When larvae are starved, their wing disks cease growth within 4 h, which indicates the existence of an efficient coupling mechanism between the growth of the soma and growth of the imaginal disks. Disk growth is inhibited by juvenile hormone in a dose-dependent manner. In the presence of the hormone the wing disks stop growing even while the larva continues to grow normally. During the last larval instar the wing imaginal disks also undergo a complex differentiation, consisting of the development of the lacunae and tracheation that define the future adult wing venation system. In normally growing larvae, differentiation of the wing disk is tightly correlated with wing size. Differentiation and size can be dissociated by starvation. If larvae are starved at any time after differentiation has begun, differentiation continues at a normal rate, even though the wing disk does not grow. Differentiation does not begin spontaneously in larvae that are starved before differentiation has begun. These findings indicate that the initiation of differentiation and its continuation are controlled independently. Juvenile hormone inhibits differentiation in a dose-dependent manner. Upon treatment with juvenile hormone, the stage of differentiation becomes fixed. These findings indicate that continued differentiation of the wing disk can only occur in the absence of juvenile hormone. Although the circulating level of juvenile hormone may be elevated during starvation, it is unlikely that this elevation is responsible for the observed effect of starvation on growth and differentiation of the disk.     

Competition among growing organs and developmental control of morphological asymmetry

Fluctuating asymmetry is often used as a measure of developmental instability, although its developmental basis is poorly understood. Theoretical models and experimental studies have suggested that feedback interactions between structures on the left and right body sides play a pivotal role in the control of asymmetry. Here we provide experimental evidence that competition for a limiting resource can generate such interactions between growing organs. In our experiments in the butterfly Precis coenia (Lepidoptera: Nymphalidae), hindwing imaginal discs were removed from one or both body sides of caterpillars. Emerging butterflies were thus missing one or both hindwings, but had heavier forewings, mid- and hindlegs than untreated controls. When only one hindwing was removed, the forewing and hindleg on the treated side were heavier than on the untreated side. The asymmetry and overall weight increase in response to wing disc removal diminished with increasing physical distance of the responding tissue from the imaginal disc removed. Our findings are consistent with the hypothesis that growing imaginal discs compete for a haemolymph-borne resource, such as a nutrient or growth factor. Such competition is a possible mechanism for feedback interactions and may thus participate in the developmental control of asymmetry.     

Worldwide patterns of mitochondrial DNA differentiation in the harbor seal (Phoca vitulina)

The harbor seal (Phoca vitulina) has one of the broadest geographic distributions of any pinniped, stretching from the east Baltic, west across the Atlantic and Pacific Oceans to southern Japan. Although individuals may travel several hundred kilometers on annual feeding migrations, harbor seals are generally believed to be philopatric, returning to the same areas each year to breed. Consequently, seals from different areas are likely to be genetically differentiated, with levels of genetic divergence increasing with distance. Differentiation may also be caused by long-standing topographic barriers such as the polar sea ice. We analyzed samples of 227 harbor seals from 24 localities and defined 34 genotypes based on 435 bp of control region sequence. Phylogenetic analysis and analysis of molecular variance showed that populations in the Atlantic and Pacific Oceans and east and west coast populations of these oceans are significantly differentiated. Within these four regions, populations that are geographically farthest apart generally are the most differentiated and often do not share genotypes or differ in genotype frequency. The average corrected sequence divergence between populations in the Atlantic and Pacific Oceans is 3.28% +/- 0.38% and those among populations within each of these oceans are 0.75% +/- 0.69% and 1.19% +/- 0.65%, respectively. Our results suggest that harbor seals are regionally philopatric, on the scale of several hundred kilometers. However, genetic discontinuities may exist, even between neighboring populations such as those on the Scottish and east English coasts or the east and west Baltic. The mitochondrial data are consistent with an ancient isolation of populations in both oceans, due to the development of polar sea ice. In the Atlantic and Pacific, populations appear to have been colonized from west to east with the European populations showing the most recent common ancestry. We suggest the recent ancestry of European seal populations may reflect recolonization from Ice Age refugia after the last glaciation.      

Definition of a juvenile hormone-sensitive period in Rhodnius prolixus

This paper is an attempt to establish the times of onset and termination of the juvenile hormone-sensitive period for metamorphosis in fifth-instar larvae of Rhodnius. Small regions of the abdominal integument were exposed to discrete pulses of the juvenile hormone analogue ZR-515 (methoprene) by applying small drops of a mixture in paraffin to the dorsum at various times after a bloodmeal and removing these drops after different time intervals. The diffusion coefficient of the analogue in the integument was estimated and used together with estimates of its metabolism to determine the lag times between application of the analogue and its rise to above threshold concentration in the epidermis, and between removal of the analogue source and its fall to below threshold concentration in the epidermis. These lag times were estimated to be 1.5 and 24h, respectively. Knowledge of the lag times makes it possible to establish the limits of the juvenile hormone-sensitive period or metamorphosis from the responses of larvae to variously-timed pulses of the analogue. The juvenile hormone-sensitive period has the following properties. For the population as a whole it lasts from about day 3 to about day 9 after a bloodmeal. Any individual in that population, however, only requires the presence of juvenile hormone during a 2 to 4-day period. The exact duration of an individual's sensitive period within these limits is a stochastic event. Surprisingly, for any individual, a pulse of juvenile hormone is equally effective when experienced early as when experienced late during its juvenile hormone-sensitive period.     

Diuresis after a bloodmeal in female Anopheles freeborni

Female Anopheles freeborni discharge urine rapidly and copiously for a brief time after taking a meal of blood. This diuresis begins immediately upon cessation of feeding and continues for about 30 min at a constant rate. A decline in this rate follows and diuresis is completed by 50 min after feeding. This time-course of diuresis is independent of the size of the meal; diuresis after large meals occures at a higher rate, not over a longer time, than diuresis after small meals. Heat-stable and saline-soluble substances that induce rapid excretion by isolated Malpighian tubules can be extracted from the head and thoracic nervous system, suggesting the presence of a neurosecretory diuretic hormone similar to that found in other blood-sucking insects. Decapitation or section of the ventral nerve cord abolishes the rapid phase of diuresis after a bloodmeal. Therefore, in analogy to the situation in the tsetse fly, the head is required either as the source of a diuretic hormone or as link in the pathway that stimulates its release.     

Structure,haemolymph titre,and regulatory effects of juvenile homone during ovarian maturation in Leucophaea maderae

Juvenile hormone (JH) synthesized and secreted in vitro by the corpora allata of mated adult Leucophaea maderae females was determined to be JH III (methyl-10,11-epoxy-3,7,11-trimethyl-2,6-dodecadienoate).The haemolymph titre of JH was determined during maturation of the terminal oöcytes in the first reproductive cycle of L. maderae. In virgin females, JH is not detectable in the haemolymph during the first eight days following adult emergence; however, by 10 days after emergence, trace quantities of JH are apparent. Mating stimuli induce a dramatic increase in the concentration of haemolymph JH, with a peak occurring approximately 12 days after mating; thereafter, the JH concentration declines until it has reached an undetectable level 19 days after mating, at the time of chorion deposition.During ovarian maturation, changes in the rates of synthesis of vitellogenin by the fat body and DNA by the ovary correlate closely with the haemolymph titre of JH. However, no such correlation exists between the JH titre and the extensive ovarian protein synthesis that occurs in L. maderae coincident with chorion formation.The effects of JH I and JH III on both vitellogenin synthesis and ovarain DNA synthesis are statistically similar.