Postembryonic brain development in the monarch butterfly,Danaus plexippus plexippus L.

Development Genes and Evolution -     

Influence of acclimation temperature on the shivering behavior of the butterfly Danaus plexippus (L.)

Summary Most of the monarch butterflies kept at 4–5° C for a few days shivered when released at a test temperature of 15–16° C, whereas fewer of the butterflies kept at 23–24° C did so. Cold-acclimated butterflies shivered more readily, as indicated by the length of the interval between release at the test temperature and the onset of shivering, and they shivered for longer periods of time. The effects of cold acclimation were reversible, but in only 1 out of 3 replicates was the warm acclimation clearly reversed. Cool animals shivered at room temperature, indicating that body temperature and not ambient temperature is important in releasing the behavior. It is suggested that the acclimation involves alteration in the central neurons controlling the activity of muscles involved in shivering.I thank Miss Janice Ruppert and Mr. C. J. Doughty for their valuable technical assistance. The co-operation of the administrators of New Brighton Beach State Park in permitting me to collect in the park is appreciated.     

Morphology of the larval and adult brains of the monarch butterfly, Danaus plexippus plexippus, L

Cell population and neuropile morphology of larval and adult brains of the monarch butterfly, Danaus plexippus plexippus, L., are compared. The larval brain is in continuous transition, the processes of adult brain development being underway from the earliest larval stages. It is characterized by a less diverse population of cells and more homogenous fiber areas than those of the adult. Neuroblasts, which divide to form the neurones of the adult brain, occur either in discrete proliferation centers or scattered among the larval ganglion cells. The larval brain contains, in addition to small homogeneous antennal centers and a distinct larval optic center, rapidly developing adult optic centers, corpora pedunculata, and protocerebral bridge. The larval brain lacks a central body. Major differences between larval and adult brains are clearly related to the increased dependence of the adult upon sensory input from the eyes and antennae.     

Overwintering ecology of the monarch butterfly, Danaus plexippus L., in California

Abstract. 1. Population and physiological measurements were made on monarch butterflies in three overwintering colonies along the California coast during the 1975–76 season. 2. Mark, release and recapture studies indicated that two northern colonies (Muir Beach and Santa Cruz) stabilized at maxima of about 40000 and 95 000 individuals with little movement in or out of colonies, whereas the southern one at Santa Barbara appeared dynamic with a maximum of about 45 000. 3. Initially low, sexual activity builds to a massive mating ceremony in February, followed by rapid colony dispersal. The butterflies deteriorate gradually during the overwintering period, but males do so to a greater extent because of their involvement in frenzied courting flights. 4. Changes in several parameters at the Muir Beach colony were measured between 1 November and 2 February. Right forewing length did not change with time, indicating no differential mortality with respect to size. However, both sexes lost about 25% of their total dry weight, and 49–60% of their lipid contents. Minimum fat levels were about 20 mg and some butterflies were as lean on arrival as were the most depleted overwintering survivors. 5. The importance of nectaring in restoring lipid reserves is potentially great but remains uninvestigated. The hypothesis is put forward that the distance spring migrants are capable of flying is dependent largely upon their lipid reserves following the overwintering period. This and the intracolony mass-mating ceremony are seen as randomizing factors which prevent geographic differentiation and allow the maintenance of the generalized, nature of the migration, which in turn enables the monarch to exploit the vast milkweed flora across the North American continent. 6. Cardenolides also diminished during the wintering, but to a substantially lesser extent than the lipids. There was no evidence for a change in the proportion of cardenolide-positive butterflies, suggesting no differential elimination by predation of the poison-free palatable individuals. This is the first field evidence in support of the theory of automimicry. 7. The overwintering behaviour of the monarch butterfly is seen as an integrative force in optimizing foodplant exploitation, reproduction, anti-predator, and migration strategies.     

Effect of photoperiod and temperature on reproduction of the monarch butterfly,Danaus plexippus

Optimal posteclosion ovarian development in monarch butterflies occurs at about 28°C. Higher and lower temperatures appear progressively inhibitory. Reproductive gland development in males follows a similar pattern. Juvenile hormone injections stimulate oöcyte maturation in animals held at 20 and 35°C but are most effective at the higher temperature. Juvenile hormone injections are ineffective at 10 and 15°C. The stimulatory effect of increasing photophase on ovarian development was most apparent at optimal temperature. The possible significance of our findings to monarch migration is discussed.     

Distribution of the monarch butterfly, Danaus plexippus (L.) (Lepidoptera: Nymphalidae), in western North America

The standard model for the migration of the monarch butterfly in western North America has hitherto been movement in the autumn to overwintering sites in coastal California, followed by a return inland by most individuals in the spring. This model is based largely on observational and limited tagging and recovery data. In this paper we test the model by plotting many years of museum and collection records on a monthly basis on a map of the region. Our plots suggest a movement of Oregon, Washington and other north-western populations of summer butterflies to California in the autumn, but movement of more north-easterly populations (e.g. from Idaho and Montana) along two pathways through Nevada, Utah and Arizona to Mexico. The more westerly of these two pathways may follow the Colorado River south as indicated by museum records and seasonal temperature data. The eastern pathway may enter northern Utah along the western scarp of the Wasatch Mountains and run south through Utah and Arizona. Further analysis of distributions suggests that monarch butterflies in the American West occur primarily along rivers, and there are observations indicating that autumn migrants often follow riparian corridors. More data are needed to test our new model; we suggest the nature of the data required.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 85 , 491–500.     

Consistent differences among individual monarch butterflies (Danaus plexippus L.) in flight activity at low temperatures

Previous work suggested differences in allozyme frequencies between samples of monarch butterflies collected at different times of the day. This study examines the prediction that some individuals are active consistently earlier in the day than others. Four specific hypotheses were tested: (1) Individuals caught early on one day were more likely than those caught late to be caught early on subsequent days. This was true in some experiments. (2) Individuals caught early in the field were more likely than those caught late to fly early in outdoor flight cages. This was true when outdoor temperatures were low (between 10 and 20^oC) but not when they were above 20^oC. (3) Individuals caught early in the field were more likely to be able to fly at low body temperatures than those caught late. This was not true. (4) Individuals that flew early in outdoor flight cages were more likely to be able to fly at low body temperatures. This was not true. Overall we conclude, on the basis of field and flight cage experiments, that some individuals are active consistently earlier in the day than others. The lack of a relationship between field and temperature cabinet results indicates that this is not purely a result of differences in ability to fly at low body temperatures. The implications of these results are discussed.     

Migration of the monarch butterfly, Danaus plexippus: energy sources,

The origin and nature of the metabolic reserves of the monarch butterfly during its autumn migration through Missouri were examined. Information was obtained about the butterfly's protein, glycogen, lipid, and lipid-soluble pigment composition. The principal results showed that lipids, which made up ca. 45 per cent of the insect's dry weight, were the major reserve. Analyses of the lipid classes showed that triglycerides were the dominant class in each sex. Separate analyses of the thorax and abdomen showed that >90 per cent of the lipids were present in the massive fat body which is restricted to the abdomen. The adult female rapidly incorporated U-¹⁴C-d-glucose into abdominal glycerides. The rôle of larval reserves and nectar consumption in migration and ovarial maturation was discussed.     

An experimental investigation of patch use by female monarch butterflies,Danaus plexippus (L.)

Monarch butterflies, which breed throughout the year in southeastern Queensland, Australia, were studied in four dense milkweed patches during the winter months (June-August) 1983. The percentage of marked females recaptured was measured in each of four 15-day sampling periods. In patches where males were experimentally removed, female recapture rate decreased compared with patches which had the same density reduction but no change in sex ratio. There was a significant correlation between female recapture rate and the proportion of males in samples, but female recapture rate was not correlated with population density, the number of males, the number of females or the proportion of young butterflies in the samples. We propose that females assessed patch quality by the sex ratios, and left a patch sooner if encounter rate with males was low. Since males provide a nutrient ‘reward’ during copulation, they may be a limited resource for females during winter. An alternative interpretation, that females left a patch sooner when female density was higher, was not supported as strongly by the data.     

The structure and function of the bursa copulatrix of the monarch butterfly (Danaus plexippus)

The bursa compulatrix of the Monarch butterfly was investigated utilizing light microscopy, histochemistry, and scanning and transmission electron microscopy in order to relate its morphology to the release of sperm from the spermatophore. The bursa has a row of large chitinous teeth on either side of the organ. The dorsal and ventral surfaces are covered with chitinous plates, the plates having bristles on one side. A single layer of cells lies under both the plates and teeth, one columnar cell under each plate, one cuboidal cell under each tooth. The toothed area has no muscle cells. However, the dorsal and ventral hemispheres of the bursa each have a crescent-shaped packet of muscle fibers that traverse the organ; there are no longitudinal fibers. Spermatophores with thick walls were found in the bursal lumen. Morphological evidence suggests that the presence of the spermatophores is sensed by the bristles and that the packets are opened by contraction of the muscles bringing the large teeth into contact with the spermatophore wall.     

Contact chemoreception related to host selection and oviposition behaviour in the monarch butterfly, Danaus plexippus

Abstract. Behavioural events during host selection by ovipositing monarch butterflies (Danaus plexippus (L.), Danainae, Nymphalidae) include tapping the leaf surface with fore-tarsi and touching this surface with mid-tarsi (‘drumming’) and antennae. Flavonoids identified from host plant extracts are known to stimulate oviposition. Scanning electron microscopy revealed the presence of contact-chemoreceptor sensilla on all appendages that contact the leaf surface. This electrophysiological study was conducted to identify the contact chemoreceptors that are sensitive to the known oviposition stimuli and are therefore probably involved in host recognition. Receptor cells of conspicuous sensilla grouped in clusters on fore-tarsi of females were sensitive to the behaviourally active butanol fraction of host plant (Asclepias curassavica) extract. However, these receptors generally had low sensitivity to three oviposition-stimulating flavonoids identified from this fraction, but they were also sensitive to the butanol fraction of a non-host (Brassica oleracea). Chemoreceptors in sensilla of the tarsomers 2–4 of the mid-legs also responded to the behaviourally active fraction of host plant extract and showed some sensitivity to two of the flavonoids that stimulate oviposition. Similar results were obtained from receptor cells in sensilla on the tip of the antennae. Most of these sensilla had cells responding to the butanol fraction of A. curassavica extract but only 25% of them were also sensitive to one of the behaviourally active flavonoids. These electrophysiological results, in combination with behavioural observations, suggest that host selection in monarch butterflies relies on a complex pattern of peripheral sensory information from several types of tarsal and antennal contact chemoreceptors.     

Differences in phenotypic traits and migratory strategies between eastern North American monarch butterflies,Danaus plexippus (L.)

The eastern North American population of the monarch butterfly (Danaus plexippus plexippus) has different migratory routes. The majority fly to overwintering colonies in Mexico and others take an eastern route through Florida and Cuba. Monarchs migrating through Florida–Cuba do not overwinter and are mostly found nectaring and flying close to vegetation. This present study explores whether Florida–Cuba versus Mexican migrants differ in (1) phenotypic traits important for migration (e.g. wing size and condition, lipid and lean mass content, and reproductive status) and (2) migratory strategies. The monarch natal grounds (e.g. migrants versus residents) were determined through thin‐layer chromatography cardenolide fingerprint and stable isotopes (hydrogen δ²H and carbon δ¹³C). In addition, wing size and condition, lipid and lean mass, and reproductive status were determined. The results suggest that Mexican migrants are better suited for longer sustained flights and successful overwinter periods as a result of larger wings in better condition, reproductive diapause, and significant fat content. By contrast, Florida–Cuba migrants are more suited for shorter flights and opportunistic migratory strategies, given that their wings were in poor condition, as well as the active reproductive status of > 50% of these butterflies and their significantly low fat content. Eastern monarch migration is more complex and diverse than previously assumed. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ?? , ??–??.     

Monosaccharide compositions of Danaus plexippus (monarch butterfly) and Trichoplusia ni (cabbage looper) egg glycoproteins

Monosaccharide compositions of eggs from Danaus plexippus (monarch butterfly) and Trichoplusia ni (cabbage looper) were analyzed. Analyses were performed mainly with high performance anion exchange chromatography (HPAEC) using crude extracts of eggs or SDS-PAGE separated and PVDF-blotted protein bands. Man and GlcN were the major components in all cases, but low levels of Gal and Fuc were possibly present in some samples. Some T. ni egg glycoproteins even contained GalN. Although a peak comigrating with Neu5Ac could be detected with HPAEC-PAD or RP-HPLC (fluorometry) after derivatization with 1,2-diamino-4,5-methylenedioxy-benzene, the quantities were too small to be significant as an integral part of the analyzed glycoproteins. These data suggests that most of glycans on the glycoproteins are pauci-Man type N-glycans, but a small portion of N-glycan may be either hybrid type or complex type.     

Genetic differentiation between Australian and North American populations of the monarch butterfly Danaus plexippus (L.) (Lepidoptera: Nymphalidae): an exploration using allozyme electrophoresis

Allozyme analysis was used to address the question of the source of the Australian populations of the monarch butterfly Danaus plexippus (L.). The study had three major aims: (1) To compare the levels of diversity of Australian and Hawaiian populations with potential source populations. (2) To determine whether eastern and western North American populations were sufficiently divergent for the Australian populations to be aligned to a source population. (3) To compare the differentiation among regions in Australia and North America to test the prediction of greater genetic structure in Australia, as a consequence of reduced migratory behaviour. The reverse was found, with F _ST values an order of magnitude lower in Australia than in North America. Predictably, Australian and Hawaiian populations had lower allelic diversity, but unexpected higher heterozygosity values than North American populations. It was not possible to assign the Australian populations to a definitive source, although the high levels of similarity of Australian populations to each other suggest a single colonization event. The possibility that the Australian populations have not been here long enough to reach equilibrium is discussed. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 75 , 437–452.     

Migratory connectivity of the monarch butterfly (Danaus plexippus): patterns of spring re-colonization in eastern North America

Each year, millions of monarch butterflies (Danaus plexippus) migrate up to 3000 km from their overwintering grounds in central Mexico to breed in eastern North America. Malcolm et al. (1993) articulated two non-mutually exclusive hypotheses to explain how Monarchs re-colonize North America each spring. The 'successive brood' hypothesis proposes that monarchs migrate from Mexico to the Gulf Coast, lay eggs and die, leaving northern re-colonization of the breeding range to subsequent generations. The 'single sweep' hypothesis proposes that overwintering monarchs continue to migrate northward after arriving on the Gulf coast and may reach the northern portion of the breeding range, laying eggs along the way. To examine these hypotheses, we sampled monarchs throughout the northern breeding range and combined stable-hydrogen isotopes (δD) to estimate natal origin with wing wear scores to differentiate between individuals born in the current vs. previous year. Similar to Malcolm et al. (1993), we found that the majority of the northern breeding range was re-colonized by the first generation of monarchs (90%). We also estimated that a small number of individuals (10%) originated directly from Mexico and, therefore adopted a sweep strategy. Contrary to Malcolm et al. (1993), we found that 62% of monarchs sampled in the Great Lakes originated from the Central U.S., suggesting that this region is important for sustaining production in the northern breeding areas. Our results provide new evidence of re-colonization patterns in monarchs and contribute important information towards identifying productive breeding regions of this unique migratory insect.     

Spatial metabolomics reveal divergent cardenolide processing in the monarch (Danaus plexippus) and the common crow butterfly (Euploea core)

Although being famous for sequestering milkweed cardenolides, the mechanism of sequestration and where cardenolides are localized in caterpillars of the monarch butterfly (Danaus plexippus, Lepidoptera: Danaini) is still unknown. While monarchs tolerate cardenolides by a resistant Na⁺/K⁺-ATPase, it is unclear how closely related species such as the nonsequestering common crow butterfly (Euploea core, Lepidoptera: Danaini) cope with these toxins. Using novel atmospheric-pressure scanning microprobe matrix-assisted laser/desorption ionization mass spectrometry imaging, we compared the distribution of cardenolides in caterpillars of D. plexippus and E. core. Specifically, we tested at which physiological scale quantitative differences between both species are mediated and how cardenolides distribute across body tissues. Whereas D. plexippus sequestered most cardenolides from milkweed (Asclepias curassavica), no cardenolides were found in the tissues of E. core. Remarkably, quantitative differences already manifest in the gut lumen: while monarchs retain and accumulate cardenolides above plant concentrations, the toxins are degraded in the gut lumen of crows. We visualized cardenolide transport over the monarch midgut epithelium and identified integument cells as the final site of storage where defences might be perceived by predators. Our study provides molecular insight into cardenolide sequestration and highlights the great potential of mass spectrometry imaging for understanding the kinetics of multiple compounds including endogenous metabolites, plant toxins, or insecticides in insects.     

Spring remigration of the monarch butterfly, Danaus plexippus (Lepidoptera: Nymphalidae) in north-central Florida: estimating population parameters using mark-recapture

Monarch butterflies [Danaus plexippus) of the eastern North American population migrate each fall from die northern U.S.A. and southern Canada to overwintering sites in Mexico and return the following spring to the southeastern U.S.A. where they lay eggs and then die. The spring remigration is the least studied phase in the annual migration cycle. We therefore conducted a mark-recapture study and examined population recolonization dynamics and residence time in a north-central Florida pasture where the monarch's milkweed host plant (Asclepias humistrata) was abundant. Beginning in late March 1995 two waves of monarchs arrived, their numbers peaked at 71 individuals by mid-April, and the butterflies disappeared in early May. After arriving, the adults remained for 3–5 days, laid eggs and then continued to migrate. We also compared population sizes and arrival times in 1994 and 1996. We found no evidence of a second spring generation, which was also consistent with the deteriorating quality of the A. humistrata plants. Individuals of the new spring generation disappear shordy after eclosion. The arriving population was approximately nine times greater in 1995 than in 1996. Our findings support two recent hypodieses: (1) the bird-like migration of the monarch butterfly in North America evolved with the northward expansion and phenology of milkweeds; and (2) monarchs appear to be migratory throughout their annual cycle of several generations. By lingering for only a short time at each milkweed patch they encounter, the old monarchs returning from Mexico locate the resurgent milkweed flora over an extensive area in the southern states. Then, within less than a month, their fresh offspring continue the migration and exploit the unfolding cornucopia of milkweeds as the spring advances northward. The more we discover about the biology of this insect, the more remarkable is its annual migratory, breeding and overwintering cycle.     

Phenology of weight, moisture and energy reserves of Australian monarch butterflies, Danaus plexippus

Abstract. 1. In the Sydney area of New South Wales, dry weights of reproductive monarch butterflies averaged 156 mg and were higher during winter than in other seasons. Dry weights of non-reproductive monarchs ranged from 216 to 324 mg and declined by 15–25% during over wintering.
2. Fat in reproductive butterflies ranged from 0.009 to 0.017g/0.1g dry weight and was lower during winter than in other seasons. Fat content of non-reproductive monarchs was higher (0.019–0.037 g/0.1g) and declined by 24–51% during over wintering.
3. Lean dry weights of reproductive monarchs were lower than those of non-reproductive individuals. Lean dry weights of non-reproductive butterflies increased rapidly at the beginning of over wintering and remained high throughout the winter. Analysis of protein content indicated the higher lean weight of non-reproductive monarchs was due to greater protein levels.
4. Moisture content of monarchs did not vary with season or reproductive status and appeared to be correlated with ambient humidity.
5. Non-reproductive monarch butterflies in New South Wales adjust bio chemically during over wintering. Energy reserve dynamics of these butterflies are comparable to those that occur in non-reproductive monarchs in North America.