Development of specific RIA and ELISA to study trypsin modulating oostatic factor in mosquitoes.
Trypsin modulating oostatic factor (TMOF), a decapeptide (H-YDPAPPPPPP-OH) that signals the termination of trypsinlike enzyme biosynthesis in the mosquito midgut, was covalently bound to Keyhole Limpet Hemocyanin using N-hydroxysuccinimide and dicyclohexylcarbodiimide. Polyclonal antibodies raised in rabbits against this conjugate were used to develop specific RIA and enzyme linked immunosorbent assay (ELISA) to detect the peptide hormone in female Aedes aegypti. TMOF and its analogs TMOF(B) (H-DYPAPPPPPP-OH), P4 (H-YDPAPPPP-OH), P1 (H-YDPAP-OH), and poly-L-proline were tested with the antiserum. The antiserum fully recognized TMOF and partially recognized P4. Using both RIA and ELISA, we report that the amount of TMOF in the mosquito ovary is 100 +/- 20 ng (S.E.) and 96 +/- 1.4 ng (S.E.), respectively, for each assay. Minute quantities of TMOF a thousandfold lower than in the ovary were found in the mosquito brain, indicating that the hormone is probably not neural but ovarian in origin.
Borovsky
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University of Florida-IFAS, Florida Medical Entomology Laboratory, Vero Beach 32962.
Powell
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CA
Carlson
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DA
D000330:Aedes / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D004064:Digestive System / Q000201:enzymology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D007814:Larva; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis* / Q000096:biosynthesis; D010053:Ovary / Q000737:chemistry; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods; D014357:Trypsin / Q000096:biosynthesis*
D009842:Oligopeptides; C065612:Peptide oostatic hormone; D014357:Trypsin