H. Frederik Nijhout

Development and evolution of adaptive polyphenisms

SUMMARY Phenotypic plasticity is the primitive character state for most if not all traits. Insofar as developmental and physiological processes obey the laws of chemistry and physics, they will be sensitive to such environmental variables as temperature, nutrient supply, ionic environment, and the availability of various macro‐ and micronutrients. Depending on the effect this phenotypic plasticity has on fitness, evolution may proceed to select either for mechanisms that buffer or canalize the phenotype against relevant environmental variation or for a modified plastic response in which some ranges of the phenotypic variation are adaptive to particular environments. Phenotypic plasticity can be continuous, in which case it is called a reaction norm, or discontinuous, in which case it is called a polyphenism. Although the morphological discontinuity of some polyphenisms is produced by discrete developmental switches, most polyphenisms are due to discontinuities in the environment that induce only portions of what is in reality a continuous reaction norm. In insect polyphenisms, the environmental variable that induces the alternative phenotype is a token stimulus that serves as a predictor of, but is not itself, the environment to which the polyphenism is an adaptation. In all cases studied so far, the environmental stimulus alters the endocrine mechanism of metamorphosis by altering either the pattern of hormone secretion or the pattern of hormone sensitivity in different tissues. Such changes in the patterns of endocrine interactions result in the execution of alternative developmental pathways. The spatial and temporal compartmentalization of endocrine interactions has produced a developmental mechanism that enables substantial localized changes in morphology that remain well integrated into the structure and function of the organism.

A THRESHOLD SIZE FOR METAMORPHOSIS IN THE TOBACCO HORNWORM, MANDUCA SEXTA (L.)

1. Under standard laboratory rearing conditions, caterpillars of Manduca sexta invariably go through 5 larval instars before pupating.2. Evidence is presented that the head capsule width of a given instar is proportional to the weight that the individual had attained at the time that the molt to that instar occurred. Fifth-instar larvae with a large range of head capsule sizes were produced by temporarily starving 3rd and 4th instars, thus inducing them to molt at subnormal weights.3. Further observations on such larvae revealed that individuals with head capsules wider than 5.1 mm proceeded to pupate at the following molt whereas larvae with smaller head capsules underwent a supernumerary larval molt.4. It was concluded that larvae of Manduca simply continue to grow and molt until they reach or exceed a sharply defined threshold size (corresponding to a head capsule size of 5.1 mm). The instar in which this threshold size is attained is then the final larval instar during which the corpora allata will be...

optix Drives the Repeated Convergent Evolution of Butterfly Wing Pattern Mimicry

Heliconius butterfly wing pattern mimicry is driven by cis-regulatory variation of the optix gene. Mimicry—whereby warning signals in different species evolve to look similar—has long served as a paradigm of convergent evolution. Little is known, however, about the genes that underlie the evolution of mimetic phenotypes or to what extent the same or different genes drive such convergence. Here, we characterize one of the major genes responsible for mimetic wing pattern evolution in Heliconius butterflies. Mapping, gene expression, and population genetic work all identify a single gene, optix, that controls extreme red wing pattern variation across multiple species of Heliconius. Our results show that the cis-regulatory evolution of a single transcription factor can repeatedly drive the convergent evolution of complex color patterns in distantly related species, thus blurring the distinction between convergence and homology.

Pattern formation on lepidopteran wings: Determination of an eyespot

Color patterns on lepidopteran wings are believed to be organized around a series of hypothetical foci. Foci presumably serve as sources of positional information, directing synthesis of appropriate pigments in their vicinity. Species-specific color patterns arise by variations in the number of foci on the wing and variations in the rules by which positional information is interpreted. The present paper provides experimental evidence for the existence of a focus that determines the large eyespot on the forewing of the Buckeye butterfly,Precis coenia. Cautery of a small group of about 300 cells at the center of the presumptive eyespot, early during wing development, can completely inhibit development of the eyespot. The same group of cells can be transplanted to another region of the wing and induce ring-shaped pigment patterns in the host tissue around the graft. These findings show that a focus is a physiological entity. Color pattern induction is an intrinsic property of a small group of cells. Presumably these cells produce a factor that affects the production of enzymes involved in pigment synthesis.

A mathematical model of the folate cycle--new insights into folate homeostasis

A mathematical model is developed for the folate cycle based on standard biochemical kinetics. We use the model to provide new insights into several different mechanisms of folate homeostasis. The model reproduces the known pool sizes of folate substrates and the fluxes through each of the loops of the folate cycle and has the qualitative behavior observed in a variety of experimental studies. Vitamin B12 deficiency, modeled as a reduction in the Vmax of the methionine synthase reaction, results in a secondary folate deficiency via the accumulation of folate as 5-methyltetrahydrofolate (the “methyl trap”). One form of homeostasis is revealed by the fact that a 100-fold up-regulation of thymidylate synthase and dihydrofolate reductase (known to occur at the G1/S transition) dramatically increases pyrimidine production without affecting the other reactions of the folate cycle. The model also predicts that an almost total inhibition of dihydrofolate reductase is required to significantly inhibit the thymidylate synthase reaction, consistent with experimental and clinical studies on the effects of methotrexate. Sensitivity to variation in enzymatic parameters tends to be local in the cycle and inversely proportional to the number of reactions that interconvert two folate substrates. Another form of homeostasis is a consequence of the nonenzymatic binding of folate substrates to folate enzymes. Without folate binding, the velocities of the reactions decrease approximately linearly as total folate is decreased. In the presence of folate binding and allosteric inhibition, the velocities show a remarkable constancy as total folate is decreased.

The Physiological Basis of Reaction Norms: The Interaction Among Growth Rate, the Duration of Growth and Body Size

Abstract The general effects of temperature and nutritional quality on growth rate and body size are well known. We know little, however, about the physiological mechanisms by which an organism translates variation in diet and temperature into reaction norms of body size or development time. We outline an endocrine-based physiological mechanism that helps explain how this translation occurs in the holometabolous insect Manduca sexta (Sphingidae). Body size and development time are controlled by three factors: (i) growth rate, (ii) the timing of the cessation of juvenile hormone secretion (measured by the critical weight) and (iii) the timing of ecdysteroid secretion leading to pupation (the interval to cessation of growth [ICG] after reaching the critical weight). Thermal reaction norms of body size and development time are a function of how these three factors interact with temperature. Body size is smaller at higher temperatures, because the higher growth rate decreases the ICG, thereby reducing the amount of mass that can accumulate. Development time is shorter at higher temperatures because the higher growth rate decreases the time required to attain the critical weight and, independently, controls the duration of the ICG. Life history evolution along altitudinal, latitudinal and seasonal gradients may occur through differential selection on growth rate and the duration of the two independently controlled determinants of the growth period.

The biological significance of substrate inhibition: a mechanism with diverse functions

Many enzymes are inhibited by their own substrates, leading to velocity curves that rise to a maximum and then descend as the substrate concentration increases. Substrate inhibition is often regarded as a biochemical oddity and experimental annoyance. We show, using several case studies, that substrate inhibition often has important biological functions. In each case we discuss, the biological significance is different. Substrate inhibition of tyrosine hydroxylase results in a steady synthesis of dopamine despite large fluctuations in tyrosine due to meals. Substrate inhibition of acetylcholinesterase enhances the neural signal and allows rapid signal termination. Substrate inhibition of phosphofructokinase ensures that resources are not devoted to manufacturing ATP when it is plentiful. In folate metabolism, substrate inhibition maintains reactions rates in the face of substantial folate deprivation. Substrate inhibition of DNA methyltransferase serves to faithfully copy DNA methylation patterns when cells divide while preventing de novo methylation of methyl‐free promoter regions.

Bombyxin is a growth factor for wing imaginal disks in Lepidoptera

The mechanisms that control the growth rate of internal tissues during postembryonic development are poorly understood. In insects, the growth rate of imaginal disks varies with nutrition and keeps pace with variation in somatic growth. We describe here a mechanism by which the growth of wing imaginal disks is controlled. When wing imaginal disks of the butterfly Precis coenia are removed from the larva and placed in a standard nutrient-rich tissue culture medium they stop growing, suggesting that nutrients alone are not sufficient to support normal growth. Such disks can be made to grow at a normal rate by supplementing the culture medium with an optimal concentration of the steroid hormone 20-hydroxyecdysone and with hemolymph taken from growing larvae. The growth-promoting activity of the hemolymph is caused by a heat-stable factor that can be extracted from the CNS and appears to be identical to the neurohormone bombyxin, a member of the insulin family of proteins. Synthetic bombyxin stimulates growth at concentrations as low as 30 ng/ml, and specific antibodies to bombyxin completely remove growth-promoting activity from the hemolymph. Bombyxin evidently acts together with 20-hydroxyecdysone to stimulate cell division and growth of wing imaginal disks. It appears that the level of bombyxin in the hemolymph is modulated by the brain in response to variation in nutrition and is part of the mechanism that coordinates the growth of internal organs with overall somatic growth.

GENETICS OF FLUCTUATING ASYMMETRY: A DEVELOPMENTAL MODEL OF DEVELOPMENTAL INSTABILITY

Although numerous studies have found that fluctuating asymmetry (FA) can have a heritable component, the genetic and developmental basis of FA is poorly understood. We used a developmental model of a trait, according to a diffusion‐threshold process, whose parameters are under genetic control. We added a small amount of random variation to the parameter values of this model to simulate developmental noise. As a result of the nonlinearity of the model, different genotypes differed in their sensitivity to developmental noise, even though the noise is completely random and independent of the genotype. The heritable component of FA can thus be understood as genetically modulated expression of variation that is itself entirely nongenetic. The loci responsible for this genetic variation of FA are the same that affect the left/right mean of the trait, showing that genetic variation for FA does not require genes that specifically control FA. Furthermore, the model offers alternative explanations for phenomena widely discussed in the literature on FA, for instance, the correlations between FA and heterozygosity and between FA and trait size. The model underscores the importance of dominance and epistasis, and therefore unites the study of FA with the classical theory of quantitative genetics.

Trade‐offs during the Development of Primary and Secondary Sexual Traits in a Horned Beetle

Resource allocation trade‐offs during development affect the final sizes of adult structures and have the potential to constrain the types and magnitude of evolutionary change that developmental processes can accommodate. Such trade‐offs can arise when two or more body parts compete for a limited pool of resources to sustain their growth and differentiation. Recent studies on several holometabolous insects suggest that resource allocation trade‐offs may be most pronounced in tissues that grow physically close to each other. Here we examine the nature and magnitude of developmental trade‐offs between two very distant body parts: head horns and genitalia of males of the horned scarab beetle Onthophagus taurus. Both structures develop from imaginal disklike tissues that undergo explosive growth during late larval development but differ in exactly when they initiate their growth. We experimentally ablated the precursor cells that normally give rise to male genitalia at several time points during late larval development and examined the degree of horn development in these males compared to that of untreated and sham‐operated control males. We found that experimental males developed disproportionately larger horns. Horn overexpression was weakest in response to early ablation and most pronounced in males whose genital disks were ablated just before larvae entered the prepupal stage. Our results suggest that even distant body parts may rely on a common resource pool to sustain their growth and that the relative timing of growth may play an important role in determining whether, and how severely, growing organs will affect each other during development. We use our findings to discuss the physiological causes and evolutionary consequences of resource allocation trade‐offs.