Prothoracicostatic Activity of the Ecdysis-Regulating Neuropeptide Crustacean Cardioactive Peptide (CCAP) in the Desert Locust
Abstract
:1. Introduction
2. Results
2.1. Fifth Instar Morphology in Correlation to the Ecdysteroid Peak and Structure of the Integument
2.2. Identification, Molecular and Functional Characterization of the S. gregaria CCAP Precursor and Receptors
2.3. Localization and Developmental Expression Patterns of the S. gregaria CCAP Precursor and Receptors
2.4. Knockdown of Ecdysteroid Receptor Components Influences Expression of Schgr-CCAPR Transcripts in the Prothoracic Gland
2.5. Phenotypic and Molecular Effects of CCAP Precursor or Receptor RNAi-Mediated Knockdown
2.5.1. Knockdown Efficiency of the dsCCAP(pre/R) Constructs
2.5.2. RNAi Ecdysis Phenotypes
2.5.3. Effects on Halloween Gene Expression in the Prothoracic Glands
2.6. Effect of CCAP on Ecdysteroidogenesis in the Prothoracic Glands
3. Discussion
3.1. Physical Characteristics Can Be Used to Identify Distinct Points within the Molt Cycle
- The observed rise in interwing distance (IW) correlates with the onset of the molting cycle, approximately marking the peak in circulating ecdysteroids and a significant increase in exuvial space compared to day 4 in the fifth nymphal stage (N5D4).
- The weight peak (WP) coincides with a noticeable and significant increase in the exuvial space compared to N5IW.
- The weight decrease (WD) is associated with an additional noticeable increase in exuvial space compared to N5WP, as well as significant shrinkage of the old endocuticle.
3.2. The Locust’s CCAP Signaling System Is Structurally and Functionally Conserved
3.3. The Imperative Role of the CCAP Signaling System in Ecdysis Is Conserved in S. gregaria
3.4. Identification of a Prothoracicostatic Function of the CCAP Signaling System in S. gregaria
4. Materials and Methods
4.1. Insect Rearing
4.2. Sampling Points Based on Locust Morphology
4.3. 20E Enzyme Immunoassay
4.4. Correlating Locust Morphology to Ecdysteroid Titer
4.5. Histology
4.6. Cell-Based Reporter Assays
4.7. Cloning, Sequence Analysis and Phylogeny of Schgr-CCAPpre and Schgr-CCAPRs
4.8. RNA Interference Experiments
4.9. RNA Extraction, cDNA Synthesis and qRT-PCR
4.10. In Vitro Incubation of Prothoracic Glands
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Verbakel, L.; Lenaerts, C.; Abou El Asrar, R.; Zandecki, C.; Bruyninckx, E.; Monjon, E.; Marchal, E.; Vanden Broeck, J. Prothoracicostatic Activity of the Ecdysis-Regulating Neuropeptide Crustacean Cardioactive Peptide (CCAP) in the Desert Locust. Int. J. Mol. Sci. 2021, 22, 13465. https://doi.org/10.3390/ijms222413465
Verbakel L, Lenaerts C, Abou El Asrar R, Zandecki C, Bruyninckx E, Monjon E, Marchal E, Vanden Broeck J. Prothoracicostatic Activity of the Ecdysis-Regulating Neuropeptide Crustacean Cardioactive Peptide (CCAP) in the Desert Locust. International Journal of Molecular Sciences. 2021; 22(24):13465. https://doi.org/10.3390/ijms222413465
Chicago/Turabian StyleVerbakel, Lina, Cynthia Lenaerts, Rania Abou El Asrar, Caroline Zandecki, Evert Bruyninckx, Emilie Monjon, Elisabeth Marchal, and Jozef Vanden Broeck. 2021. "Prothoracicostatic Activity of the Ecdysis-Regulating Neuropeptide Crustacean Cardioactive Peptide (CCAP) in the Desert Locust" International Journal of Molecular Sciences 22, no. 24: 13465. https://doi.org/10.3390/ijms222413465