Releasing Digestive Enzymes by Brain Peptides in the Larvae of Pieris brassicae L. (Lepidoptera: Pieridae)

  • Ali Sharifloo
  • Arash ZibaeeEmail author
  • Jalal Jalali Sendi
  • Khalil Talebi Jahroumi
  • Abdolrashid Janghorbani
Research Article


Neuropeptides synthesized by insect neurosecretory cells are engaged in several physiological process like immunity, feeding and reproduction so they deserve to be undertaken in subsequent experiments for discovery and manipulation regarding applied objectives. Due to importance of insect ability to utilize plant tissues, the present study was conducted to extract brain peptides in the fifth instar larvae of Pieris brassicae via High performance liquid chromatography and to identify their role in induction of digestive enzyme syntheses. Five peaks were determined and named as fractions 1, 2, 3, 4 and 5. The midguts of starved fifth larval instars were treated by each fraction of brain extract in vitro. The highest activities of α-amylase and lipase were found in the midgut treated by fractions 4 and 1, respectively while the fraction number 5 imposed the highest activities of glucosidases. Fraction 2 led to the highest activities of trypsin, chymotrypsin and elastase as the given serine proteinases as well as the two exopeptidases namely amino- and carboxypeptidases. Moreover, midgut treatments by different concentrations of TLCK (N-p-Tosyl-l-lysine methyl ester hydrochloride), TPCK (Tosyl phenylalanyl chloromethyl ketone), AEBSF (4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride) and Acarbose significantly decreased the activities of α-amylase, trypsin, chymotrypsin and elastase by the calculated IC50 values of 0.287, 0.439, 0.288 and 0.361 mM, respectively. The findings demonstrated that different neuropeptides would be engaged in enzymatic release of P. brassicae hence, manipulation of peptide receptors can change physiological process of insects and opens a selective strategy to pest control.


Pieris brassicae Brain peptide Digestive enzyme High performance liquid chromatography Manipulation of peptide receptors 



The authors would like to appreciate assistances of Ms. Minoo Sajjadian for HPLC analysis. The present study was supported by University of Guilan, Iran.

Compliance with Ethical Standards

Conflict of interest

The authors declare that there is no conflict of interest among the authors regarding publication of this manuscript.

Ethical Standard Statement

The experiments was conducted based on the rules on compliance with ethical statements.


  1. 1.
    Harshini S, Reshmi V, Sreekumar S (2003) A brain peptide stimulates release of amylase from the midgut tissue of larvae of Opisina arenosella Walk. (Lepidoptera: Cryptophasidae). Neuropeptides 37(3):133–139CrossRefGoogle Scholar
  2. 2.
    Chapman RF (2013) The insects: structure and function, 5th edn. Cambridge University Press, CambridgeGoogle Scholar
  3. 3.
    Duve H, Thorpe A (1981) Gastrin/cholecystokinin-like immunoreactivity neurons in the brain of the blow fly Calliphora erythrocephala (Diptera). Gen Comp Endocrinol 43:381–391CrossRefGoogle Scholar
  4. 4.
    Duve H, Thorpe A (1984) Immunocytochemical mapping of gastrin/cholecystokinin-like peptides in the neuroendocrine system of the blow fly Calliphora erythrocephala (Diptera). Cell Tissue Res 237:309–320CrossRefGoogle Scholar
  5. 5.
    Yui R, Fujita T, Ito S (1980) Insulin-, gastrin-, pancreatic polypeptide-like immunoreactivity neurons in the brain of the silk worm, Bombyx mori. Biomed Res 1:42–46CrossRefGoogle Scholar
  6. 6.
    Verhaert P, De Loof A (1985) Substance P-like immunoreactivity in the central nervous system of the blattarian insect Periplanata americana revealed by a monoclonal antibody. Histochemistry 83:501–5077CrossRefGoogle Scholar
  7. 7.
    Harshini S, Nachman RJ, Sreekumar S (2002) In vitro release of digestive enzymes by FMRF amide related neuropeptides and analogues in the lepidopteran insect Opisina arenosella (Walk.). Peptides 23:1759–1763CrossRefGoogle Scholar
  8. 8.
    Karuppaiah V, Sujithra M (2013) Insect neuropeptides: as potential targets for pest control, research news for U (RNFU) ISSN: 2250–3668, vol 10Google Scholar
  9. 9.
    Zibaee A (2012) A digestive lipase of Pieris brassicae L. (Lepidoptera: Pieridae): purification, characterization and host plants effects. Arch Insect Biochem Physiol 81:1–19CrossRefGoogle Scholar
  10. 10.
    Zibaee A (2012) A digestive lipase of Pieris brassicae L. (Lepidoptera: Pieridae): purification, characterization and host plants effects. Arch Insect Biochem Physiol 81(1):1–19CrossRefGoogle Scholar
  11. 11.
    Sajjadian SM, Hosseininaveh V, Talebi Jahromi K (2014) Some brain peptides regulating the secretion of digestive enzymes in the Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae). J Plant Prot Res 54:279–286CrossRefGoogle Scholar
  12. 12.
    Frugoni JAC (1957) Tampone universale di Britton e Robinson a forza ionica costante. Gazz Chim Ital 87:403–407Google Scholar
  13. 13.
    Bernfeld P (1955) Amylases alpha and beta. Methods Enzymol 1:149–158CrossRefGoogle Scholar
  14. 14.
    Tsujita T, Ninomiya H, Okuda H (1989) p-nitrophenyl butyrate hydrolyzing activity of hormone sensitive lipase from bovine adipose tissue. J Lipid Res 30:997–1004Google Scholar
  15. 15.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenolreagent. J Biol Chem 193:265–275Google Scholar
  16. 16.
    Harshini S, Sreekumar S (2001) Isolation and partial purification of the digestive enzyme-release peptide hormone from the midgut of larvae of Opisina arenosella Walk. (Lepidoptera: Cryptophasidae. In: Muraleedharan K et al (eds) Advances in entomology. Association for Advancement of Entomology, Trivandrum, pp 191–197Google Scholar

Copyright information

© The National Academy of Sciences, India 2017

Authors and Affiliations

  1. 1.Department of Plant Protection, Faculty of Agricultural SciencesUniversity of GuilanRashtIran
  2. 2.Department of Plant Protection, College of Agriculture and Natural ResourcesUniversity of TehranKarajIran

Personalised recommendations