Moth Agrotis Ipsilon Research Articles

Is the rapid post-mating inhibition of pheromone response triggered by ecdysteroids or other factors from the sex accessory glands in the male moth Agrotis ipsilon?

In many animals, male copulation is dependent on the detection and processing of female-produced sex pheromones, which is generally followed by a sexual refractory post-ejaculatory interval (PEI). In the male moth, Agrotis ipsilon, this PEI is characterized by a transient post-mating inhibition of behavioral and central nervous responses to sex pheromone, which prevents males from re-mating until they have refilled their reproductive tracts for a potential new ejaculate. However, the timing and possible factors inducing this rapid olfactory switch-off are still unknown. Here, we determined the initial time delay and duration of the PEI. Moreover, we tested the hypothesis that the brain, the testis and/or the sex accessory glands (SAGs) could produce a factor inducing the PEI. Lastly, we investigated the possible involvement of ecdysteroids, hormones essential for development and reproduction in insects, in this olfactory plasticity. Using brain and SAG cross-injections in virgin and newly-mated males, surgical treatments, wind tunnel behavioral experiments and EIA quantifications of ecdysteroids, we show that the PEI starts very shortly after the onset of copulation, and that SAGs contain a factor, which is produced/accumulated after copulation to induce the PEI. Moreover, SAGs were found to be the main source of ecdysteroids, whose concentration decreased after mating, whereas it increased in the haemolymph. 20-Hydroxyecdysone (20E) was identified as the major ecdysteroid in SAGs of A. ipsilon males. Finally, 20E injections did not reduce the behavioral pheromone response of virgin males. Altogether our data indicate that 20E is probably not involved in the PEI.

Steroid hormone signaling is involved in the age-dependent behavioral response to sex pheromone in the adult male moth Agrotis ipsilon

In most animals, including insects, male reproduction depends on the detection and processing of female-produced sex pheromones. In the male moth, Agrotis ipsilon, both behavioral response and neuronal sensitivity in the primary olfactory center, the antennal lobe (AL), to female sex pheromone are age- and hormone-dependent. In many animal species, steroids are known to act at the brain level to modulate the responsiveness to sexually relevant chemical cues. We aimed to address the hypothesis that the steroidal system and in particular 20-hydroxyecdysone (20E), the main insect steroid hormone, might also be involved in this olfactory plasticity. Therefore, we first cloned the nuclear ecdysteroid receptor EcR (AipsEcR) and its partner Ultraspiracle (AipsUSP) of A. ipsilon, the expression of which increased concomitantly with age in ALs. Injection of 20E into young sexually immature males led to an increase in both responsiveness to sex pheromone and amount of AipsEcR and AipsUSP in their ALs. Conversely, the behavioral response decreased in older, sexually mature males after injection of cucurbitacin B (CurB), an antagonist of the 20E/EcR/USP complex. Also, the amount of AipsEcR and AipsUSP significantly declined after treatment with CurB. These results suggest that 20E is involved in the expression of sexual behavior via the EcR/USP signaling pathway, probably acting on central pheromone processing in A. ipsilon.

Molecular identification and differential expression of sensory neuron membrane proteins in the antennae of the black cutworm moth Agrotis ipsilon

The insect sensory neuron membrane proteins (SNMPs) SNMP1 and SNMP2 are transmembrane domain-containing proteins and are homologs of the vertebrate CD36 transmembrane proteins. It has been suggested that SNMPs play a significant role in insect chemoreception. Previous studies have demonstrated that SNMP1 is expressed in the pheromone-sensitive olfactory receptor neurons (ORNs), whereas SNMP2 is expressed in the supporting cells. In this study, we identified two full-length SNMP transcripts, AipsSNMP1 and AipsSNMP2, in the black cutworm moth Agrotis ipsilon (Hufnagel). The qRT-PCR results indicated that the AipsSNMP1 and AipsSNMP2 transcripts were expressed significantly higher in the antennae than in other tissues of both sexes. The expression of AipsSNMP1 and AipsSNMP2 in the antennae from different development stages of both sexes was investigated and was shown to begin to express in the pupae stage from 3days before emergence and then increased dramatically at the day of the emergence, and the high expression levels were maintained during the following 4days after the emergence in both sexes. The mating status had no effect on the expression levels of the AipsSNMP1 and AipsSNMP2 transcripts. Consistent with previous in situ hybridization studies in other Lepidoptera insects, our immunolocalization results at protein level demonstrated that both AipsSNMP1 and AipsSNMP2 were expressed in pheromone-sensitive sensilla trichodea but with a completely different expression profile. AipsSNMP1 is more uniformed and highly expressed along the membrane of the ORN dendrites, whereas AipsSNMP2 is widely distributed at the bottom of the sensilla trichodea and highly localized in the sensillum lymph. Our studies provide further detailed evidence for the involvement and general functional role of insect SNMPs in the detection of sex pheromones and general odorant molecules.

Sex pheromone recognition and immunolocalization of three pheromone binding proteins in the black cutworm moth Agrotis ipsilon

Insect pheromone binding proteins (PBPs) are believed to solubilize and transport hydrophobic sex pheromones across sensillum lymph to membrane-associated pheromone receptors. To address the molecular mechanisms of PBPs in insect pheromone perception, we undertook a systemic study on the PBPs of the black cutworm Agrotis ipsilon at transcript as well as protein level from tissue distribution and cellular localization to pheromone binding affinity. We cloned three full-length PBP genes AipsPBP1-3 from A. ipsilon antennae, and demonstrated that AipsPBP1–3 transcripts were highly expressed in male antennae. The electron microscopic examinations revealed at least six types of olfactory sensilla on male and female antenna: trichodea, chaetica, basiconica, coeloconica, squamiformia and Böhm bristles. The immunocytochemistry results demonstrated that AipsPBP1-3 proteins were strongly expressed in the sensillum lymph of the trichoid sensilla of male moth. The binding assays showed that AipsPBP1 had high binding affinities with the major sex pheromone components Z7-12:Ac and Z9-14:Ac among five related chemicals and was clustered together with the long trichoid sensilla-expressing LdisPBPs of Lymantria dispar. AipsPBP2 showed high binding affinities also with Z11-16:Ac. AipsPBP3 displayed a high affinity only with Z11-16:Ac. Our studies provide further detail evidences for the involvement of moth PBPs in pheromone discrimination and selective recognition of specific components of the female sex pheromone blends.

Plant odour stimuli reshape pheromonal representation in neurons of the antennal lobe macroglomerular complex of a male moth

Male moths are confronted with complex odour mixtures in a natural environment when flying towards a female-emitted sex pheromone source. Whereas synergistic effects of sex pheromones and plant odours have been observed at the behavioural level, most investigations at the peripheral level have shown an inhibition of pheromone responses by plant volatiles, suggesting a potential role of the central nervous system in reshaping the peripheral information. We thus investigated the interactions between sex pheromone and a behaviourally active plant volatile, heptanal, and their effects on responses of neurons in the pheromone-processing centre of the antennal lobe, the macroglomerular complex, in the moth Agrotis ipsilon. Our results show that most of these pheromone-sensitive neurons responded to the plant odour. Most neurons responded to the pheromone with a multiphasic pattern and were anatomically identified as projection neurons. They responded either with excitation or pure inhibition to heptanal, and the response to the mixture pheromone + heptanal was generally weaker than to the pheromone alone, showing a suppressive effect of heptanal. However, these neurons responded with a better resolution to pulsed stimuli. The other neurons with either purely excitatory or inhibitory responses to all three stimuli did not exhibit significant differences in responses between stimuli. Although the suppression of the pheromone responses in AL neurons by the plant odour is counter-intuitive at first glance, the observed better resolution of pulsed stimuli is probably more important than high sensitivity to the localization of a calling female.

Differential Interactions of Sex Pheromone and Plant Odour in the Olfactory Pathway of a Male Moth

Most animals rely on olfaction to find sexual partners, food or a habitat. The olfactory system faces the challenge of extracting meaningful information from a noisy odorous environment. In most moth species, males respond to sex pheromone emitted by females in an environment with abundant plant volatiles. Plant odours could either facilitate the localization of females (females calling on host plants), mask the female pheromone or they could be neutral without any effect on the pheromone. Here we studied how mixtures of a behaviourally-attractive floral odour, heptanal, and the sex pheromone are encoded at different levels of the olfactory pathway in males of the noctuid moth Agrotis ipsilon. In addition, we asked how interactions between the two odorants change as a function of the males' mating status. We investigated mixture detection in both the pheromone-specific and in the general odorant pathway. We used a) recordings from individual sensilla to study responses of olfactory receptor neurons, b) in vivo calcium imaging with a bath-applied dye to characterize the global input response in the primary olfactory centre, the antennal lobe and c) intracellular recordings of antennal lobe output neurons, projection neurons, in virgin and newly-mated males. Our results show that heptanal reduces pheromone sensitivity at the peripheral and central olfactory level independently of the mating status. Contrarily, heptanal-responding olfactory receptor neurons are not influenced by pheromone in a mixture, although some post-mating modulation occurs at the input of the sexually isomorphic ordinary glomeruli, where general odours are processed within the antennal lobe. The results are discussed in the context of mate localization.

Mating-induced differential coding of plant odour and sex pheromone in a male moth

Innate behaviours in animals can be influenced by several factors, such as the environment, experience, or physiological status. This behavioural plasticity originates from changes in the underlying neuronal substrate. A well-described form of plasticity is induced by mating. In both vertebrates and invertebrates, males experience a post-ejaculatory refractory period, during which they avoid new females. In the male moth Agrotis ipsilon, mating induces a transient inhibition of responses to the female-produced sex pheromone. To understand the neural bases of this inhibition and its possible odour specificity, we carried out a detailed analysis of the response characteristics of the different neuron types from the periphery to the central level. We examined the response patterns of pheromone-sensitive and plant volatile-sensitive neurons in virgin and mated male moths. By using intracellular recordings, we showed that mating changes the response characteristics of pheromone-sensitive antennal lobe (AL) neurons, and thus decreases their sensitivity to sex pheromone. Individual olfactory receptor neuron (ORN) recordings and calcium imaging experiments indicated that pheromone sensory input remains constant. On the other hand, calcium responses to non-pheromonal odours (plant volatiles) increased after mating, as reflected by increased firing frequencies of plant-sensitive AL neurons, although ORN responses to heptanal remained unchanged. We suggest that differential processing of pheromone and plant odours allows mated males to transiently block their central pheromone detection system, and increase non-pheromonal odour detection in order to efficiently locate food sources.

Post-mating sexual abstinence in a male moth

In most animals, male copulation is dependent on the detection and processing of female-produced sex pheromones. In males, a refractory post-ejaculatory interval (PEI) follows copulation, allowing them to avoid direct remating until they have replenished their reproductive tracts. In the moth Agrotis ipsilon, newly-mated males show a transient inhibition of behavioral and central nervous responses to sex pheromone. Using non-pheromonal (plant) odours, pheromones, and their mixture, we now show that the observed lack of pheromone response originates from differential post-mating odour processing in the brain. Although mated males still respond to plant odors alone, their response to mixtures depends on the added pheromone concentration. Below a specific threshold, sex pheromone is not detected at the brain level; above this threshold, it becomes inhibitory. This PEI can thus be interpreted as a « refusal to respond », which contradicts the generally accepted paradigm of sleep-like/exhaustion behaviour during PEI.

Cloning of an octopamine/tyramine receptor and plasticity of its expression as a function of adult sexual maturation in the male moth Agrotis ipsilon

In the male moth Agrotis ipsilon behavioural response and antennal lobe (AL) neuron sensitivity to the female-produced sex pheromone increase with age and juvenile hormone (JH) level. We recently showed that the neuromodulator, octopamine (OA), interacts with JH in this age-dependent olfactory plasticity. To further elucidate its role, we cloned a full cDNA encoding a protein that presents biochemical features essential to OA/tyramine receptor (AipsOAR/TAR) function. The AipsOAR/TAR transcript was detected predominantly in the antennae, the brain and, more specifically, in ALs where its expression level varied concomitantly with age. This expression plasticity indicates that AipsOAR/TAR might be involved in central processing of the pheromone signal during maturation of sexual behaviour in A. ipsilon.

Quantitative analysis of sex-pheromone coding in the antennal lobe of the moth Agrotis ipsilon: a tool to study network plasticity

To find a mating partner, moths rely on pheromone communication. Released in very low amounts, female sex pheromones are used by males to identify and localize females. Depending on the physiological state (i.e. age, reproductive state), the olfactory system of the males of the noctuid moth Agrotis ipsilon is 'switched on or off'. To understand the neural basis of this behavioural plasticity, we performed a detailed characterization of the qualitative, quantitative and temporal aspects of pheromone coding in the primary centre of integration of pheromonal information, the macroglomerular complex (MGC) of the antennal lobe. MGC neurons were intracellularly recorded and stained in sexually mature virgin males. When stimulating antennae of males with the three main components of the female pheromone blend, most of the neurons showed a biphasic excitatory-inhibitory response. Although they showed different preferences, 80% of the neurons responded at least to the main pheromone component (Z-7-dodecenyl acetate). Six stained neurons responding to this component had their dendrites in the largest MGC glomerulus. Changes in the stimulus intensity and duration affected the excitatory phase but not the inhibitory phase properties. The stimulus intensity was shown to be encoded in the firing frequency, the number of spikes and the latency of the excitatory phase, whereas the stimulus duration only changed its duration. We conclude that the inhibitory input provided by local interneurons following the excitatory phase might not contribute directly to the encoding of stimulus characteristics. The data presented will serve as a basis for comparison with those of immature and mated males.

Three-dimensional antennal lobe atlas of the male moth, Agrotis ipsilon: a tool to study structure-function correlation.

The glomerular structure of the primary olfactory neuropil has long been thought to play an important role in odour coding. In insects, the number of glomeruli in the antennal lobe is limited in most species to fewer than 100 compared with more than 1,000 in vertebrates, making it possible to identify individual glomeruli. A complete three-dimensional atlas of the glomeruli within the antennal lobe of the male noctuid moth Agrotis ipsilon was constructed. All 66 glomeruli were singly identifiable in both antennal lobes of the three brains investigated. Further, six antennal lobes containing intracellularly stained projection neurones were reconstructed. By using the atlas, the respective target glomerulus of each projection neurone could be identified. The importance of the glomerular atlas as a tool to study central olfactory processing and its plasticity is discussed.

Transient post–mating inhibition of behavioural and central nervous responses to sex pheromone in an insect

Mating is costly for both male and female insects and should therefore only occur if it is likely to be successful. Within one scotophase, which is the dark period of the light cycle, male moths can only produce one single spermatophore, which is transferred to the female during mating. Remating within the same scotophase would thus be unsuccessful. We tested the hypothesis that newly mated males of the moth Agrotis ipsilon have developed an energy-saving strategy based on the transient inhibition of their sexual behaviour, thus avoiding unsuccessful remating. Agrotis ipsilon males do not copulate more than once during the same scotophase. Moreover, newly mated males do not respond behaviourally to the female sex pheromone although electroantennograms showed that their peripheral olfactory system is fully functional. However, intracellular recordings of antennal lobe neurons showed that the sensitivity for the synthetic sex pheromone blend decreased as compared with that of unmated males. Both the sexual behaviour and the sensitivity of the antennal lobe neurons were restored when tested during the next scotophase. Our results show a fast, transient neuronal plasticity that 'switches off' the olfactory system, which could prevent males from mating unsuccessfully.

Molecular cloning and structural analysis of 3-hydroxy-3-methylglutaryl coenzyme A reductase of the moth Agrotis ipsilon.

The enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, which plays a key role in isoprenoid biosynthesis, catalyses the synthesis of mevalonate from HMG-CoA. Insects do not synthesize cholesterol de novo, rather mevalonate derivatives lead to non-sterol isoprenoids which are essential for development and reproduction. In this paper, we describe an HMG-CoA reductase of the moth Agrotis ipsilon and we report its expression in fat body, ovary, muscle, brain and corpora allata tissues of adult specimens. The analysis of the cDNA reveals that it encodes a polypeptide of 833 amino acids (Mr = 89785). Alignments of this HMG-CoA reductase from A. ipsilon with the homologous sequences of other eukaryotes shows a high degree of conservation in all species studied. Parsimony analysis based on these alignments produced dendrograms congruent with the current systematic schemes. This suggests that, during eukaryote evolution, HMG-CoA reductase diversified in parallel with taxonomic splitting.

Molecular cloning and tissue expression of an insect farnesyl diphosphate synthase.

The enzyme farnesyl-diphosphate synthase (FPS, EC2.5.1.1/EC2.5.1.10), which has been shown to play a key role in isoprenoid biosynthesis, catalyzes the synthesis of farnesyl diphosphate from isopentenyl diphosphate and di-methylallyl diphosphate. Insects do not synthesize cholesterol de novo, rather farnesyl diphosphate leads to the formation of nonsterol isoprenoids, which are essential for insect development and reproduction. In this paper, we describe the characterization of one FPS from the moth Agrotis ipsilon, the first insect FPS to be reported. An homologous probe was obtained through a nested PCR strategy using degenerate primers designed from the conserved domains of FPS from other organisms. The complete cDNA clone was isolated by PCR screening of a brain cDNA library by using homologous primers deduced from the probe. Analysis of the nucleotide sequence revealed that the cDNA encodes a polypeptide of 412 amino acids (Mr = 47 170), which shares regions similar to the FPS of other organisms, but exhibits singularities such as an extra N-terminal extension of approximately 70 amino acid residues. Using an RNase protection assay, a protected fragment corresponding to the region encoding the FPS catalytic site was found in brain, ovary, fat body and corpora allata samples, but not in muscle. FPS is overexpressed in the corpora allata, the endocrine gland that produces the juvenile hormones. These hormones are specific to insects and play a crucial role in regulating insect physiology.

Effect of juvenile hormone on the central nervous processing of sex pheromone in an insect.

Behavioral sex pheromone responsiveness in the male moth Agrotis ipsilon was previously shown to be controlled by juvenile hormone (JH). However, this morphogenetic hormone did not change the sensitivity of antennae to sex pheromones. To analyze the possible involvement of JH in the central integration of the female-produced sex pheromone, we investigated the pheromone response of olfactory antennal lobe (AL) interneurons in male A. ipsilon as a function of age and JH status by using intracellular recordings. When the antennae were stimulated with the sex pheromone blend, the sensitivity of olfactory AL neurons increased with age, as does the JH-dependent behavioral and physiological development of A. ipsilon males. Furthermore, males surgically deprived of JH showed a significant decrease in the sensitivity of the AL neurons. JH injection in operated or in young males restored or induced, respectively, a high sensitivity of the AL neurons. JH seems likely to be involved in the plasticity of the adult insect brain by modulating the central nervous processing of olfactory information, thus allowing mate recognition and reproduction at the optimal time.

A cDNA, from Agrotis ipsilon, that encodes the pheromone biosynthesis activating neuropeptide (PBAN) and other FXPRL peptides

A cDNA encoding the prohormone of the pheromone biosynthesis activating neuropeptide (PBAN) in the moth Agrotis ipsilon was isolated. The cDNA contains 834 nucleotides, coding for a 193-amino acid protein that exhibits 89% identity with PBAN prohormones of other moths. The prohormone contains five potential peptides belonging to the FXPRL family. The peptide corresponding to the Bombyx mori diapause hormone exhibits an extra residue, and the C-terminal leucine is replaced by an isoleucine, introducing a new type of variability in this family of peptides. Northern blot analysis revealed expression in suboesophagal ganglion complexes. Constitutive heterologous expression of Agi-PBAN cDNA in yeast, using three different antibodies, did not produce PBAN-immunoreactive material.

Biosynthetic activity of corpora allata, growth of sex accessory glands and mating in the male moth agrotis ipsilon (Hufnagel)

The involvement of both juvenile hormone acid (JHA) and the sex accessory glands (SAGs) in the reproduction of the male moth Agrotis ipsilon was studied as a function of age and mating status. Total protein content analysis followed by gel electrophoresis of the SAGs, radiochemical assay for JHA biosynthesis and surgical and behavioural experiments were performed. Both the protein content of the SAGs and the biosynthetic activity of the corpora allata (CA) increased with age. Allatectomy and JHA/JH treatments showed that the protein content of the SAGs is linked with the activity of the CA. The protein content of the glands, but not the rate of JHA biosynthesis, decreased just after mating, and both increased sharply 24 h later. Injection of fluvastatin, an inhibitor of JH biosynthesis, in males immediately after mating prevented the increase in JHA synthesis and lowered the total protein content of the SAGs. Moreover, fluvastatin disrupted normal spermatophore transfer during the next mating of the injected males. Our results show that JHA controls the reproduction of A. ipsilon males by its separate actions on the sex accessory glands and on sexual behaviour.

Biological Activities of Natural Sesquiterpene Lactones and the Effect of Synthetic Sesquiterpene Derivatives on Insect Juvenile Hormone Biosynthesis

Three natural sesquiterpene lactones have been assayed for their biological activity on locust (Locusta migratoria) nymphs. Since results obtained in vivo pointed to disruptions on juvenile hormone (JH)-regulated physiology, we tested the sesquiterpene lactones and a family of synthetic sesquiterpene derivatives for their ability to disturb the rate of JH biosynthesis by locust corpora allata (CA) in vitro. All the active compounds in vitro share a basic sesquiterpene double-ring structure as well as substituents with different chemical complexities. Compounds that shared an eudesmane base with different oxidation states on C3 and C12 carbon atoms were tested as JH biosynthesis inhibitors. Supplementation of the locust corpora allata culture media with exogenous JH precursors together with data obtained with corpora allata from a moth (Agrotis ipsilon) suggested that epoxidase is a target of the inhibition process.

119 Differential localization of the two putative sodium channel α-subunits in the optic lobe and the stellate ganglion of the squid Loligo breekeri

The antennal lobe (AL) of the Noctuid moth Agrotis ipsilon has emerged as an excellent model for studying olfactory processing and its plasticity in the central nervous system. Odor-evoked responses of AL neurons and input-to-output transformations involved in pheromone processing are well characterized in this species. However, the intrinsic electrical properties responsible of the firing of AL neurons are poorly known. To this end, patch-clamp recordings in current- and voltage-clamp mode from neurons located in the two main clusters of cell bodies in the ALs were combined with intracellular staining on A. ipsilon males. Staining indicated that the lateral cluster (LC) is composed of 85% of local neurons (LNs) and 15% of projection neurons (PNs). The medial cluster (MC) contains only PNs. Action potentials were readily recorded from the soma in LNs and PNs located in the LC but not from PNs in the MC where recordings showed small or no action potentials. In the LC, the spontaneous activity of about 20% of the LNs presented irregular bursts while being more regular in PNs. We also identified a small population of LNs lacking voltage-gated Na+ currents and generating spikelets. We focused on the firing properties of LNs since in about 60% of LNs, but not in PNs, action potentials were followed by depolarizing afterpotentials (DAPs). These DAPs could generate a second action potential, so that the activity was composed of action potential doublets. DAPs depended on voltage, Ca2+-channels and possibly on Ca2+-activated non-specific cationic channels. During steady state current injection, DAPs occurred after each action potential and did not require high-frequency firing. The amplitude of DAPs increased when the interspike interval was small, typically within bursts, likely arising from a Ca2+ build up. DAPs were more often found in bursting than in non-bursting LNs but do not support bursting activity. DAPs and spike doublets also occurred during odor-evoked activity suggesting that they can mediate olfactory integration in the AL.

Reproductive Maturity, Mating Status, and Long-Duration Flight Behavior of Agrotis ipsilon (Lepidoptera: Noctuidae) and the Conceptual Misuse of the Oogenesis'Flight Syndrome by Entomologists

The relationship between long-duration flight and the state of oogenesis in the migratory moth Agrotis ipsilon (Hufnagel) was examined in unmated females 1,2,3,4, and 5 d old by using computer-interfaced flight mills. Evidence presented suggests that such long-duration flights (i.e., continuous flights ≥1 h) are indices of migratory flight. There were no significant correlations between the number of chorionated eggs and flight duration, times of flight initiation and termination, or flight speed within age classes 1–4. In 5-d-old moths, however, there was a positive correlation with flight duration. Mated females began their long-duration flights ≍3 h later in the night and averaged 2 h of flight less than unmated females. Their propensity and capacity to engage in such flights, however, was still considerable. The long-duration flight behavior of 3-d-old mated males was similar to that of unmated males, as was their propensity to engage in such flights. Therefore, a delay in oogenesis and suppression of reproductive behavior are not prerequisites for the initiation of long-duration flight activity in A. ipsilon , as would be predicted if the oogenesis-flight syndrome was characteristic of this species. Our data are consistent with previous findings that suggest that most females arrive in the midwestern United States in the spring already mated. We argue that the concept of an oogenesis-flight syndrome in migratory insects has no inherent explanatory power and its only valid use is as a convenient descriptive term. Its most frequent misuse as an explanatory principle is seen in the common assumption that duration of the prereproductive period determines the maximum duration of the migratory period. Empirical evidence as well as theoretical considerations indicate that it is never safe to accept this assumption without testing. To do so risks impeding progress in understanding specific systems through unwarranted extrapolation of data or through building a house of cards.