A re-evaluation of silk measurement by the cecropia caterpillar (Hyalophora cecropia) during cocoon construction reveals use of a silk odometer that is temporally regulated

The late 5th instar caterpillar of the cecropia silk moth (Hyalophora cecropia) spins a silken cocoon with a distinct, multilayered architecture. The cocoon construction program, first described by the seminal work of Van der Kloot and Williams, consists of a highly ordered sequence of events. We perform behavioral experiments to re-evaluate the original cecropia work, which hypothesized that the length of silk that passes through the spinneret controls the orderly execution of each of the discrete events of cocoon spinning. We confirm and extend by three-dimensional scanning and quantitative measurements of silk weights that if cocoon construction is interrupted, upon re-spinning, the caterpillar continues the cocoon program from where it left off. We also confirm and extend by quantitative measurements of silk weights that cecropia caterpillars will not bypass any of the sections of the cocoon during the construction process, even if presented with a pre-spun section of a cocoon spun by another caterpillar. Blocking silk output inhibits caterpillars from performing normal spinning behaviors used for cocoon construction. Surprisingly, unblocking silk output 24-hr later did not restart the cocoon construction program, suggesting the involvement of a temporally-defined interval timer. We confirm with surgical reductions of the silk glands that it is the length of silk itself that matters, rather than the total amount of silk extracted by individuals. We used scanning electron microscopy to directly show that either mono- or dual-filament silk (i.e., equal silk lengths but which vary in their total amount of silk extracted) can be used to construct equivalent cocoons of normal size and that contain the relevant layers. We propose that our findings, taken together with the results of prior studies, strongly support the hypothesis that the caterpillar uses a silk odometer to measure the length of silk extracted during cocoon construction but does so in a temporally regulated manner. We further postulate that our examination of the anatomy of the silk spinning apparatus and ablating spinneret sensory output provides evidence that silk length measurement occurs upstream of output from the spinneret

Discordant timing between antennae disrupts sun compass orientation in migratory monarch butterflies

To navigate during their long-distance migration, monarch butterflies (Danaus plexippus) use a time-compensated sun compass. The sun compass timing elements reside in light-entrained circadian clocks in the antennae. Here we show that either antenna is sufficient for proper time compensation. However, migrants with either antenna painted black (to block light entrainment) and the other painted clear (to permit light entrainment) display disoriented group flight. Remarkably, when the black-painted antenna is removed, re-flown migrants with a single, clear-painted antenna exhibit proper orientation behaviour. Molecular correlates of clock function reveal that period and timeless expression is highly rhythmic in brains and clear-painted antennae, while rhythmic clock gene expression is disrupted in black-painted antennae. Our work shows that clock outputs from each antenna are processed and integrated together in the monarch time-compensated sun compass circuit. This dual timing system is a novel example of the regulation of a brain-driven behaviour by paired organs

A rhythmic Ror

The circadian clock mechanism in mammals involves two interlocking transcriptional feedback loops. Rev-erb alpha, through its role as a transcriptional repressor, was thought to be the primary determinant of the feedback loop that regulates Bmal1 transcription. Results reported by Sato et al. in this issue of Neuron now show that the transactivator Rora acts coordinately with Rev-erb alpha and that their competing activities on the same promoter element drive the rhythm in Bmal1 transcription. This finding defines the second feedback loop in mammals

A Rhythmic Ror

AbstractThe circadian clock mechanism in mammals involves two interlocking transcriptional feedback loops. Rev-erb α, through its role as a transcriptional repressor, was thought to be the primary determinant of the feedback loop that regulates Bmal1 transcription. Results reported by Sato et al. in this issue of Neuron now show that the transactivator Rora acts coordinately with Rev-erb α and that their competing activities on the same promoter element drive the rhythm in Bmal1 transcription. This finding defines the second feedback loop in mammals

Dimorphic cocoons of the cecropia moth (Hyalophora cecropia): Morphological, behavioral, and biophysical differences.

The larvae of the giant silk moth (Hyalophora cecropia) spin strikingly dimorphic, multilayered cocoons that are either large and fluffy (baggy) or significantly smaller and tightly woven (compact). Although these cocoon-morphs share the same function (i.e., housing for pupal to adult development during overwintering), previous work has been unable to determine why cocoon dimorphism exists. We addressed this issue in cecropia moth cocoons collected along power line right-of-way habitats in Massachusetts. We first characterized the architectural differences between cocoon-morphs for all three cocoon sections (outer and inner envelopes, and the intermediate layer separating the two). We show that outer envelope structural and ultrastructural differences are what underlie dimorphism. Using a common spinning arena, we next show that the behavioral suites used to construct the outer envelopes of the two morphs are significantly different in behavioral time investment and patterning, as well as in the location of silk placement in the common spinning arena. Finally, we compared the cocoon-morphs in response to various environmental stressors to ask whether dimorphism is an adaptive response to such pressures. In contrast to compact cocoons, we find that baggy cocoons act as heat sinks and allow greater moisture permeability; differences in outer envelope architecture underlie these characteristics. These two biophysical properties could be advantageous for pupae in baggy cocoons, during unseasonably cold or dry conditions encountered during development prior to adult emergence. Our results suggest that cocoon dimorphism in the cecropia moth may provide a bet-hedging strategy for dealing with varying environmental conditions in Massachusetts and perhaps over its entire habitat range, during pupal to adult development

Dimorphic cocoons of the cecropia moth (Hyalophora cecropia): Morphological, behavioral, and biophysical differences

The larvae of the giant silk moth (Hyalophora cecropia) spin strikingly dimorphic, multilayered cocoons that are either large and fluffy (baggy) or significantly smaller and tightly woven (compact). Although these cocoon-morphs share the same function (i.e., housing for pupal to adult development during overwintering), previous work has been unable to determine why cocoon dimorphism exists. We addressed this issue in cecropia moth cocoons collected along power line right-of-way habitats in Massachusetts. We first characterized the architectural differences between cocoon-morphs for all three cocoon sections (outer and inner envelopes, and the intermediate layer separating the two). We show that outer envelope structural and ultrastructural differences are what underlie dimorphism. Using a common spinning arena, we next show that the behavioral suites used to construct the outer envelopes of the two morphs are significantly different in behavioral time investment and patterning, as well as in the location of silk placement in the common spinning arena. Finally, we compared the cocoon-morphs in response to various environmental stressors to ask whether dimorphism is an adaptive response to such pressures. In contrast to compact cocoons, we find that baggy cocoons act as heat sinks and allow greater moisture permeability; differences in outer envelope architecture underlie these characteristics. These two biophysical properties could be advantageous for pupae in baggy cocoons, during unseasonably cold or dry conditions encountered during development prior to adult emergence. Our results suggest that cocoon dimorphism in the cecropia moth may provide a bet-hedging strategy for dealing with varying environmental conditions in Massachusetts and perhaps over its entire habitat range, during pupal to adult development

Coldness Triggers Northward Flight in Remigrant Monarch Butterflies

SummaryEach fall, eastern North American monarch butterflies (Danaus plexippus) migrate from their northern range to their overwintering grounds in central Mexico [1–3]. Fall migrants are in reproductive diapause, and they use a time-compensated sun compass to navigate during the long journey south [4–6]. Eye-sensed directional cues from the daylight sky (e.g., the horizontal or azimuthal position of the sun) are integrated in the sun compass in the midbrain central complex region [7, 8]. Sun compass output is time compensated by circadian clocks in the antennae so that fall migrants can maintain a fixed flight direction south [9, 10]. In the spring, the same migrants remigrate northward to the southern United States to initiate the northern leg of the migration cycle. Here we show that spring remigrants also use an antenna-dependent time-compensated sun compass to direct their northward flight. Remarkably, fall migrants prematurely exposed to overwintering-like coldness reverse their flight orientation to the north. The temperature microenvironment at the overwintering site is essential for successful completion of the migration cycle, because without cold exposure, aged migrants continue to orient south. Our discovery that coldness triggers the northward flight direction in spring remigrants solves one of the long-standing mysteries of the monarch migration

Coldness triggers northward flight in remigrant monarch butterflies

Each fall, eastern North American monarch butterflies (Danaus plexippus) migrate from their northern range to their overwintering grounds in central Mexico [1-3]. Fall migrants are in reproductive diapause, and they use a time-compensated sun compass to navigate during the long journey south [4-6]. Eye-sensed directional cues from the daylight sky (e.g., the horizontal or azimuthal position of the sun) are integrated in the sun compass in the midbrain central complex region [7, 8]. Sun compass output is time compensated by circadian clocks in the antennae so that fall migrants can maintain a fixed flight direction south [9, 10]. In the spring, the same migrants remigrate northward to the southern United States to initiate the northern leg of the migration cycle. Here we show that spring remigrants also use an antenna-dependent time-compensated sun compass to direct their northward flight. Remarkably, fall migrants prematurely exposed to overwintering-like coldness reverse their flight orientation to the north. The temperature microenvironment at the overwintering site is essential for successful completion of the migration cycle, because without cold exposure, aged migrants continue to orient south. Our discovery that coldness triggers the northward flight direction in spring remigrants solves one of the long-standing mysteries of the monarch migration