Skip to main content
Log in

Putative circadian pacemaker cells in the antenna of the hawkmoth Manduca sexta

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

Antennal sensory neurons in the fruit fly Drosophila melanogaster express circadian rhythms in the clock gene PERIOD (PER) and appear to be sufficient and necessary for circadian rhythms in olfactory responses. Given recent evidence for daily rhythms of pheromone responses in the antenna of the hawkmoth Manduca sexta, we examined whether a peripheral PER-based circadian clock might be present in this species. Several different cell types in the moth antenna were recognized by monoclonal antibodies against Manduca sexta PER. In addition to PER-like staining of pheromone-sensitive olfactory receptor neurons and supporting cells, immunoreactivity was detected in beaded branches contacting the pheromone-sensitive sensilla. The nuclei of apparently all sensory receptor neurons, of sensilla supporting cells, of epithelial cells, and of antennal nerve glial cells were PER-immunoreactive. Expression of per mRNA in antennae was confirmed by the polymerase chain reaction, which showed stronger expression at Zeitgeber-time 15 compared with Zeitgeber-time 3. This evidence for the expression of per gene products suggests that the antenna of the hawkmoth contains endogenous circadian clocks.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price includes VAT (France)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Baker TC, Cardé RT (1979) Endogenous and exogenous factors affecting periodicities of female calling and pheromone response in Grapholita molesta. J Insect Physiol 25:943–950

    Article  Google Scholar 

  • Bell RA, Joachim FA (1976) Techniques for rearing laboratory colonies of tobacco hornworms and pink bollworms. Ann Ent Soc 69:365–373

    Google Scholar 

  • Emery IF, Noveral JM, Jamison CF, Siwicki KK (1997) Rhythms of Drosophila period gene expression in culture. Proc Natl Acad Sci USA 94:4092–4096

    Article  CAS  PubMed  Google Scholar 

  • Flecke C, Dolzer J, Krannich S, Stengl M (2006) Perfusion with cGMP analogue adapts the action potential response of pheromone-sensitive sensilla trichoidea of the hawkmoth Manduca sexta in a daytime-dependent manner. J Exp Biol 209:3898–3912

    Article  CAS  PubMed  Google Scholar 

  • Giebultowicz JM (2001) Peripheral clocks and their role in circadian timing: insights from insects. Philos Trans R Soc Lond Biol 356:1791–1799

    Article  CAS  PubMed  Google Scholar 

  • Glossop NRJ, Hardin PE (2002) Central and peripheral circadian oscillator mechanisms in flies and mammals. J Cell Sci 115:3369–3377

    CAS  PubMed  Google Scholar 

  • Hardin PE (2005) The circadian timekeeping system of Drosophila. Curr Biol 15:R714–R722

    Article  CAS  PubMed  Google Scholar 

  • Itagaki H, Conner WE (1988) Calling behavior of Manduca sexta (L.) (Lepidoptera: Sphingidae) with notes on the morphology of the female sex pheromon gland. Ann Ent Soc Am 81:798–807

    Google Scholar 

  • Keil T, Steinbrecht RA (1984) Mechanosensitive and olfactory sensilla of insects. In: King RC, Akai H (eds) Insect ultrastructure, vol 2. Plenum, New York, pp 477–515

    Google Scholar 

  • Krishnan B, Dryer SE, Hardin PE (1999) Circadian rhythms in olfactory responses of Drosophila melanogaster. Nature 400:375–378

    Article  CAS  PubMed  Google Scholar 

  • Krishnan B, Levine JD, Lynch KS, Dowse HB, Funes P, Hall JC, Hardin PE, Dryer SE (2001) A new role for cryptochrome in a Drosophila circadian oscillator. Nature 411:313–317

    Article  CAS  PubMed  Google Scholar 

  • Linn CE, Campbell MG, Poole KR, Wu W-Q, Roelofs WL (1996) Effects of photoperiod on the circadian timing of pheromone response in male Trichoplusia ni. J Insect Physiol 42:881–891

    Article  CAS  Google Scholar 

  • Loesel R, Weigel S, Braunig P (2006) A simple fluorescent double staining method for distinguishing neuronal from non-neuronal cells in the insect central nervous system. J Neurosci Methods 155:202–206

    Article  CAS  PubMed  Google Scholar 

  • Lohr C, Deitmer JW (2006) Glia. Calcium signaling in invertebrate glial cells. Glia 54:642

    Article  PubMed  Google Scholar 

  • Merlin C, François MC, Queguiner I, Maïbèche-Coisné M, Jacquin-Joly E (2006) Evidence for a putative antennal clock in Mamestra brassicae: molecular cloning and characterization of two clock genes—period and cryptochrome—in antennae. Insect Mol Biol 15:137–145

    Article  CAS  PubMed  Google Scholar 

  • Moussian B, Schwarz H, Bartoszewski S, Nüsslein-Volhard C (2005) Involvement of chitin in exoskeleton morphognesis in Drosophila melanogaster. J Morphol 264:117–130

    Article  CAS  PubMed  Google Scholar 

  • Myers EM (2003) The circadian control of eclosion. Chronobiol Int 20:775–794

    Article  CAS  PubMed  Google Scholar 

  • Page TL, Koelling E (2003) Circadian rhythm in olfactory response in the antennae controlled by the optic lobe in the cockroach. J Insect Physiol 49:697–707

    Article  CAS  PubMed  Google Scholar 

  • Plautz JD, Kaneko M, Hall JC, Kay SA (1997) Independent photoreceptive circadian clocks throughout Drosophila. Science 278:1632–1635

    Article  CAS  PubMed  Google Scholar 

  • Rosén WQ (2002) Endogenous control of circadian rhythms of pheromone production in the turnip moth Agrotis segetum. Arch Insect Biochem Physiol 50:21–30

    Article  PubMed  Google Scholar 

  • Rosén WQ, Han GB, Lofstedt C (2003) The circadian rhythm of the sex-pheromone-mediated behavioral response in the turnip moth Agrotis segetum, is not controlled at the peripheral level. J Biol Rhythms 18:402–408

    Article  PubMed  Google Scholar 

  • Sanes JR, Hildebrand JG (1976) Structure and development of antennae in a moth, Manduca sexta. Dev Biol 51:282

    Article  Google Scholar 

  • Sauman I, Reppert SM (1996) Circadian clock neurons in the silkmoth Antheraea pernyi: novel mechanisms of PERIOD protein regulation. Neuron 17:889–900

    Article  CAS  PubMed  Google Scholar 

  • Schneider NL, Stengl M (2005) Pigment-dispersing factor and GABA synchronize cells of the isolated circadian clock of the cockroach Leucophaea maderae. J Neurosci 25:5138–5147

    Article  CAS  PubMed  Google Scholar 

  • Schneider NL, Stengl M (2006a) Extracellular long-term recordings of the isolated accessory medulla, the circadian pacemaker center of the cockroach Leucophaea maderae, reveal ultradian and hint circadian rhythms. J Comp Physiol A Neuroethol Sens Neural Behav Physiol 193:35–42

    PubMed  Google Scholar 

  • Schneider NL, Stengl M (2006b) Gap junctions between accessory medulla neurons appear to synchronize circadian clock cells of the cockroach Leucophaea maderae. J Neurophysiol 95:1996–2002

    Article  PubMed  Google Scholar 

  • Sehadova H, Markova EP, Sehnal E, Takeda M (2004) Distribution of circadian clock-related proteins in the cephalic nervous system of the silkworm, Bombyx mori. J Biol Rhythms 19:466–482

    Article  CAS  PubMed  Google Scholar 

  • Silvegren G, Löfstedt C, Rosén WQ (2005) Circadian mating activity and effect of pheromone pre-exposure on pheromone response rhythms in the moth Spodoptera littoralis. J Insect Physiol 51:277–286

    Article  CAS  PubMed  Google Scholar 

  • Steel CG, Vafopoulou X (2006) Circadian orchestration of developmental hormones in the insect Rhodnius prolixus. Comp Biochem Physiol A Mol Integr Physiol 144:351–364

    Article  PubMed  Google Scholar 

  • Tanoue S, Krishnan P, Krishnan B, Dryer SE, Hardin PE (2004) Circadian clocks in antennal neurons are necessary and sufficient for olfaction rhythms in Drosophila. Curr Biol 14:638–649

    Article  CAS  PubMed  Google Scholar 

  • Weber F (1985) Postmolt cuticle growth in a cockroach: in vitro deposition of multilamellate and circadian–like layered endocuticle. Experientia 41:1632–1635

    Article  Google Scholar 

  • Wise S, Davis NT, Tyndale E, Noveral J, Folwell G, Bedian V, Emery IF, Siwicki KK (2002) Neuroanatomical studies of PERIOD gene expression in the hawkmoth, Manduca sexta. J Comp Neurol 447:366–380

    Article  CAS  PubMed  Google Scholar 

  • Zavodska R, Sehadova H, Sauman I, Sehnal F (2005) Light-dependent PER-like proteins in the cephalic ganglia of an apterygote and a pterygote insect species. Histochem Cell Biol 123:407–418

    Article  CAS  PubMed  Google Scholar 

  • Zerr DM, Hall JC, Rosbash M, Siwicki KK (1990) Circadian fluctuations of period protein immunoreactivity in the CNS and the visual system of Drosophila. J Neurosci 10:2749–2762

    CAS  PubMed  Google Scholar 

  • Zhou X, Yuan C, Guo A (2005) Drosophila olfactory response rhythms require clock genes but not pigment dispersing factor or lateral neurons. J Biol Rhythms 20:237–244

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr. Lino Saez and Dr. M.W. Young (Rockefeller University, New York, USA) for the anti-Drosophila PER antibodies and to Dr. Anthony Gotter and Dr. S.M. Reppert (Medical School, University of Massachusetts, USA) for anti-Antheraea PER antibodies. We also thank Dr. Alexander Tups (University of Otago, New Zealand) and Tobias Fromme (University of Marburg, Germany) for helpful instructions concerning the PCR technique.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Monika Stengl.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schuckel, J., Siwicki, K.K. & Stengl, M. Putative circadian pacemaker cells in the antenna of the hawkmoth Manduca sexta . Cell Tissue Res 330, 271–278 (2007). https://doi.org/10.1007/s00441-007-0471-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-007-0471-x

Keywords

Navigation