Development of a novel screening system for allatostatin receptor agonists in search of new candidate insect growth regulators

Allatostatins (ASTs) are insect neuropeptide hormones that regulate diverse physiological functions, including feeding, growth and development, and reproduction. Therefore, regulation of allatostatin receptor (AstR) activity can be an effective tool for controlling insect growth and proliferation. H...

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Bibliographic Details
Published in:Biotechnology letters 2014-07, Vol.36 (7), p.1439-1446
Main Authors: Na, Joonho, Kim, Wonju, Yoon, Bong-June
Format: Article
Language:English
Subjects:
agonists
allatostatin
Animals
Applied Microbiology
aspartate transaminase
Biochemistry
Biomedical and Life Sciences
Biotechnology
Cell Line
Control
Drug Evaluation, Preclinical - methods
dyes
Feeding
Fluorescence
growth and development
Growth regulators
Hormones
Humans
insect growth
insect growth regulators
Insecta
Insects
Juvenile Hormones - isolation & purification
Juvenile Hormones - pharmacology
Life Sciences
mammals
membrane potential
Membranes
Microbiology
Neuropeptides - pharmacology
Original Research Paper
Peptides
Pest control
Receptors
Receptors, Neuropeptide - agonists
reproduction
Screening
Statins
transfection
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Summary:Allatostatins (ASTs) are insect neuropeptide hormones that regulate diverse physiological functions, including feeding, growth and development, and reproduction. Therefore, regulation of allatostatin receptor (AstR) activity can be an effective tool for controlling insect growth and proliferation. Here, we describe a novel screening system using a mammalian cell line in which AstR is ectopically expressed, combined with fluorescence-based measurements of the membrane potential. HEK293T cells that do not express cognate receptors for AST became responsive to AST upon transfection with AstR. The response of the membrane potential to AST could be reliably detected by measuring the fluorescence of DiBAC4(3), a voltage-sensitive dye. We also discovered that overexpressing GIRK1/2 in this cell line could augment the magnitude of hyperpolarization by AST. Our screening system produces a fast and reliable readout for the efficient screening of AstR agonists.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-014-1505-6