Diapause

Diapause is the delay in development in response to regularly and recurring periods of adverse environmental conditions.^[1][2] It is considered to be a physiological state of dormancy with very specific initiating and inhibiting conditions. Diapause is a mechanism used as a means to survive predictable, unfavourable environmental conditions, such as temperature extremes, drought or reduced food availability. Diapause is most often observed in arthropods, especially insects, and in the embryos of many of the oviparous species of fish in the order Cyprinodontiformes.^[3] (Diapause does not occur in embryos of the viviparous and ovoviviparous species of Cyprinodontiformes.)

Diapause is not only induced in an organism by specific stimuli or conditions, but once it is initiated, only certain other stimuli are capable of bringing the organism out of diapause. The latter feature is essential in distinguishing diapause as a different phenomenon from other forms of dormancy such as stratification, and hibernation.

Activity levels of diapausing stages can vary considerably among species. Diapause may occur in a completely immobile stage, such as the pupae and eggs, or it may occur in very active stages that undergo extensive migrations, such as the adult Monarch butterfly, Danaus plexippus. In cases where the insect remains active, feeding is reduced and reproductive development is slowed or halted.

Phases of insect diapause

Diapause in insects is a dynamic process consisting of several distinct phases (Kostal 2006). While diapause varies considerably from one taxon of insects to another, these phases can be characterized by particular sets of metabolic processes and responsiveness of the insect to certain environmental stimuli.^[4] Diapause can occur during any stage of development in arthropods, but each species exhibits diapause in specific phases of development. Reduced oxygen consumption is typical as is reduced movement and feeding.^[5]

Comparison of Diapause Periods

The sensitive stage is the period where stimulus must occur to trigger diapause in the organism. Examples of sensitive stage/diapause periods in various insects:^[6]

• Diatraea grandiosella (Southwestern corn borer) – sensitive stage: early larva, diapause: late larval^[7]
• Sarcophaga crassipalpis – sensitive stage: early larval, diapause: pupa
• Sarcophaga argyrostoma – sensitive stage: mid to late larval, diapause: pupa
• Manduca sexta (Tobacco hornworm) – sensitive stage: late embryonic (egg) to late larval, diapause: pupa
• Lepinotarsa decemlineata – sensitive stage: early adult, diapause: late adult
• Bombyx mori (Silkworm) – sensitive stage: late embryonic (egg) to early larval, diapause: embryonic
• Lymantria dispar (Gypsy moth) – sensitive stage: late embryonic, diapause: late embryonic
• Danaus plexipus (Monarch butterfly) – sensitive stage: early adulthood diapause: adulthood

Induction

The induction phase occurs at a genetically predetermined stage of life and occurs well in advance of the environmental stress.^[4] This sensitive stage may occur within the lifetime of the diapausing individual, or in preceding generations, particularly in egg diapause.^[8] During this phase, insects are responsive to external cues called token stimuli, which trigger the switch from direct development pathways to diapause pathways. Token stimuli can consist of changes in photoperiod, thermoperiod or allelochemicals from food plants. These stimuli are not in themselves favourable or unfavourable to development, but they herald an impending change in environmental conditions.^[2]

Preparation

The preparation phase usually follows the induction phase, though insects may go directly from induction to initiation without a preparation phase.^[4] During this phase, insects accumulate and store molecules such as lipids, proteins and carbohydrates. These molecules are used to maintain the insect throughout diapause and to provide fuel for development following diapause termination. Composition of the cuticle may be altered by changing hydrocarbon composition and by adding lipids to reduce water loss making the organism resistant to desiccation.^[9] Diapausing puparia of the flesh fly Sarcophaga crassipalpis increase the amount of cuticular hydrocarbons lining the puparium, effectively reducing the ability of water to cross the cuticle.^[10]

Initiation

Photoperiod is the most important stimulus initiating diapause.^[11] The initiation phase begins when morphological development ceases.^[4] In some cases, this change may be very distinct and can involve moulting into a specific diapause stage, or be accompanied by color change. Enzymatic changes may take place in preparation for cold hardening. For example, only diapausing adults of the fire bug, Pyrrhocoris apterus, have the enzymatic complement that allows them to accumulate polyhydric alcohols, molecules that help to lower their freezing points and thus avoid freezing.^[12] Insects may also undergo behavioural changes and begin to aggregate, migrate or search for suitable overwintering sites.

Overwintering Monarch Butterflies in diapause clustering on Oyamel trees. Note that one tree is completely covered in butterflies. These butterflies were located on a preserve outside of Angangueo, Michoacán, Mexico

Maintenance

During the maintenance phase, insects experience lowered metabolism and developmental arrest is maintained.^[4] Sensitivity to certain stimuli which act to prevent termination of diapause, such as photoperiod and temperature, is increased. At this stage, insects are unresponsive to changes in the environment that will eventually trigger the end of diapause, but they grow more sensitive to these stimuli as time progresses.

Termination

In insects that undergo obligate diapause, termination may occur spontaneously, without any external stimuli.^[4] In facultative diapausers, token stimuli must occur to terminate diapause. These stimuli may include chilling, freezing or contact with water, depending on the environmental conditions being avoided. These stimuli are important in preventing the insect from terminating diapause too soon, for instance in response to warm weather in late fall. Termination may occur at the height of unfavourable conditions, such as in the middle of winter. Over time, depth of diapause slowly decreases until direct development can resume, if conditions are favourable.

Post-diapause quiescence

Diapause frequently ends prior to the end of unfavourable conditions and is followed by a state of quiescence from which the insect can arouse and begin direct development, should conditions change to become more favourable.^[4] This allows the insect to continue to withstand harsh conditions while being ready to take advantage of good conditions as rapidly as possible.

Regulation of diapause

Diapause in insects is regulated at several levels. Environmental stimuli interact with genetic pre-programming to affect neuronal signalling, endocrine pathways and eventually metabolic and enzymatic changes.

Environmental

Environmental regulators of diapause generally display a characteristic seasonal pattern. In temperate regions, photoperiod is the most reliable cues of seasonal change.^[8] Depending on the season in which diapause occurs, either short or long days can act as token stimuli. Insects may also respond to changing day length as well as relative day length. Temperature may also act as a regulating factor, either by inducing diapause or, more commonly, by modifying the response of the insect to photoperiod.^[8] Insects may respond to thermoperiod, the daily fluctuations of warm and cold that correspond with night and day, as well as to absolute or cumulative temperature. Food availability and quality may also help regulate diapause. In the desert locust, Schistocerca gregaria, a plant hormone called gibberellin stimulates reproductive development.^[13] During the dry season, when their food plants are in senescence and lacking gibberellin, the locusts remain immature and their reproductive tracts do not develop.

Neuroendocrine

The neuroendocrine system of insects consists primarily of neurosecretory cells in the brain, the corpora cardiaca, corpora allata and the prothoracic glands.^[2] There are several key hormones involved in the regulation of diapause: juvenile hormone (JH), diapause hormone (DH), and prothoracicotropic hormone (PTTH).^[14]

Prothoracicotropic hormone stimulates the prothoracic glands to produce ecdysteroids that are required to promote development.^[14] Larval and pupal diapauses are often regulated by an interruption of this connection, either by preventing release of prothoracicotropic hormone from the brain or by failure of the prothoracic glands to respond to prothoracicotropic hormone.

The corpora allata is responsible for the production of juvenile hormone (JH). In the bean bug, Riptortus pedestris, clusters of neurons on the protocerebrum called the pars lateralis maintain reproductive diapause by inhibiting JH production by the corpora allata.^[15] Adult diapause is often associated with the absence of JH, while larval diapause is often associated with its presence.

In adults, absence of JH causes degeneration of flight muscles, atrophy or cessation of development of reproductive tissues, and halts mating behaviour. The presence of JH in larvae may prevent moulting to the next larval instar, though successive stationary moults may still occur.^[16] In the corn borer, Diatraea gradiosella, JH is required for the accumulation by the fat body of a storage protein that is associated with diapause.^[17]

Diapause hormone regulates embryonic diapause in the eggs of the silkworm moth, Bombyx mori.^[18] DH is released from the subesophageal ganglion of the mother and triggers trehalase production by the ovaries. This generates high levels of glycogen in the eggs, which is converted into the polyhydric alcohols glycerol and sorbitol. Sorbitol directly inhibits the development of the embryos. Glycerol and sorbitol are reconverted into glycogen at the termination of diapause.

Tropical diapause

Diapause in the tropics is often initiated in response to biotic rather than abiotic factors.^[19] For example, food in the form of vertebrate carcasses may be more abundant following dry seasons, or oviposition sites in the form of fallen trees may be more available following rainy seasons. Also, diapause may serve to synchronize mating seasons or reduce competition, rather than to avoid unfavourable climatic conditions.

Diapause in the tropics poses several challenges to insects that are not faced in temperate zones.^[19] Insects must reduce their metabolism without the aid of cold temperatures and may be faced with increased water loss due to high temperatures. While cold temperatures inhibit the growth of fungi and bacteria, diapausing tropical insects still have to deal with these pathogens. Also, predators and parasites may still be abundant during the diapause period.

Aggregations are common among diapausing tropical insects, especially in the orders Coleoptera, Lepidoptera and Hemiptera.^[19] Aggregations may be used as protection against predation, since aggregating species are frequently toxic and predators quickly learn to avoid them. They can also serve to reduce water loss, as seen in the fungus beetle, Stenotarsus rotundus, which forms aggregations of up to 70,000 individuals, which may be eight beetles deep. Relative humidity is increased within the aggregations and beetles experience less water loss, probably due to decreased surface area to volume ratios reducing evaporative water loss.^[20]

References

1. ^ The Insects; Structure and Function, 4th Edition. R.F. Chapman, Cambridge University Press, 1998. ISBN 0521 57048, p 403.
2. ^ ^{a b c} Tauber, M.J., Tauber, C.A., Masaki, S. (1986) Seasonal Adaptations of Insects. Oxford University Press, 414 pp.
3. ^ Glen E. Collier; William J. Murphy (August 1997). A molecular phylogeny for aplocheiloid fishes (Atherinomorpha, Cyprinodontiformes): the role of vicariance and the origins of annualism. PMID 9254916. "Annual aplocheiloid killifish embryos possess a rare ability among vertebrates to enter stages of developmental arrest (diapause) when subjected to adverse environmental conditions."  PMID 9254916
4. ^ ^{a b c d e f g} Kostal, V. (2006). Eco-physiological phases of insect diapause. Journal of Insect Physiology. 52: 113–127.
5. ^ The Insects; Structure and Function, 4th Edition. R.F. Chapman, Cambridge University Press, 1998. ISBN 0521 57048, pp. 403–404
6. ^ The Insects; Structure and Function, 4th Edition. R.F. Chapman, Cambridge University Press, 1998. ISBN 0521 57048, p 404.
7. ^ Bulletin of Entomological Research (1976), 66:75–79 Cambridge University Press, Copyright © Cambridge University Press 1976, Diapause of the southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera, Pyralidae): effects of a juvenile hormone mimic: G. M. Chippendalea1 and C.-M. Yina1a1, Department of Entomology, University of Missouri, Columbia, Missouri 65201, U.S.A.
8. ^ ^{a b c} Huffaker, C.B. and Gutierrez, A.P., Eds. 1999. Ecological Entomology. John Wiley & Sons, Inc.
9. ^ Epicuticular Wax Secretion in Diapause and Non-diapause Pupae of the Bertha Army worm, Author: HEGDEKAR, B. M., Source: Annals of the Entomological Society of America, Volume 72, Number 1, 15 January 1979 , pp. 13–15(3)Publisher: Entomological Society of America
10. ^ Yoder, J.A., Denlinger, D.L., Dennis, M.W. and Kolattukudey, P.E. (1992) Enhancement of diapausing flesh fly puparia with additional hydrocarbons and evidence for alkane biosynthesis by a decarbonylation mechanism. Insect Biochemistry and Molecular Biology. 22(3): 237–243.
11. ^ The Insects; Structure and Function, 4th Edition. R.F. Chapman, Cambridge University Press, 1998. ISBN 0521 57048, p. 404
12. ^ Kostal, V., Tollarova, M. and Sula, J. (2004) Adjustments of the enzymatic complement for polyol biosynthesis and accumulation in diapausing cold-acclimated adults of Pyrrhocoris apterus. Journal of Insect Physiology. 50: 303–313.
13. ^ Ellis, P.E., Carlisle, D.B. and Osborne, D.J. (1965) Desert locusts: sexual maturation delayed by feeding on senescent vegetation. Science. 149(3683): 546–547.
14. ^ ^{a b} Denlinger, D.L. (2002) Regulation of diapause. Annual Review of Entomology. 47: 93–122.
15. ^ Shimokawa, K., Numata, H. and Shiga, S. (2008) Neurons important for the photoperiodic control of diapause in the bean bug, Riptortus pedestris. Journal of Comparative Physiology. 194:751–762.
16. ^ Yin, C.M. and Chippendale, G.M. (1979) Diapause of the Southwestern Corn Borer, Diatraea grandiosella: further evidence showing juvenile hormone to be the regulator. Journal of Insect Physiology. 25: 513–523.
17. ^ Brown, J.J. and Chippendale, G.M. (1978) Juvenile hormone and a protein associated with the larval diapause of the Southwestern Corn Borer, Diatraea grandiosella. Insect Biochemistry. 8: 359–397.
18. ^ Horie, Y., Kanda, T. and Mochida, Y. (2000) Sorbitol as an arrester of embryonic development in diapausing eggs of the silkworm, Bombyx mori. Journal of Insect Physiology. 46: 1009–1016.
19. ^ ^{a b c} Denlinger, D.L. (1986) Dormancy in tropical insects. Annual Review of Entomology. 31:239–264.
20. ^ Yoder, J.A., Denlinger, D.L. and Wolda, H. (1992) Aggregation promotes water conservation during diapause in the tropical fungus beetle, Stenotarsus rotundus. Entomologia Experimentalis et Applicata. 63: 203–205.