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Molecular cloning and immunolocalization of a diuretic hormone receptor in rice brown planthopper ( Nilaparvata lugens )
Author(s) -
Price D. R. G.,
Du J.,
Dinsmore A.,
Gatehouse J. A.
Publication year - 2004
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.0962-1075.2004.00507.x
Subject(s) - brown planthopper , biology , malpighian tubule system , receptor , complementary dna , hemolymph , microbiology and biotechnology , biochemistry , gene , midgut , botany , larva
RNA extracted from guts of rice brown planthopper, Nilaparvata lugens , was used to clone cDNA predicted to encode a diuretic hormone receptor (DHR). The DHR, a member of the calcitonin/secretin/corticotropin‐releasing factor family of G‐protein‐coupled receptors, contains seven transmembrane domains and a large N‐terminal extracellular domain potentially involved in hormone binding. The N‐terminal domain was expressed as a recombinant protein, purified and used to raise antibodies. Anti‐DHR IgG bound specifically to Malpighian tubules in immunolocalization experiments using dissected guts, and to a putative DHR polypeptide from N. lugens gut on Western blots. Anti‐DHR IgG delivered orally to insects was not detected in the haemolymph, and showed no binding to gut or tubules, confirming that DHR N‐terminal hormone‐binding domain is not exposed to the gut lumen.