Isolation and characterization of the cDNA encoding DH(31) in the kissing bug, Rhodnius prolixus.
Rhodnius prolixus undergoes a period of rapid diuresis after ingesting large blood meals. Neurohormones with either diuretic or anti-diuretic activity control diuresis by acting on several tissues including the Malpighian tubules. One of the neurohormones that potentially plays a role in diuresis is diuretic hormone 31 (DH(31)) which belongs to the insect calcitonin-like family of diuretic hormones. Here we determine the complete cDNA sequences of three Rhopr-DH(31) splice variants (Rhopr-DH(31)-A, Rhopr-DH(31)-B and Rhopr-DH(31)-C) and characterize their expression in unfed fifth-instar R. prolixus. Reverse transcriptase-PCR demonstrates that Rhopr-DH(31) is predominantly expressed in the central nervous system (CNS) of unfed fifth-instars. However, the expression of the three splice variants differs with Rhopr-DH(31)-B expression being the highest followed by Rhopr-DH(31)-A and Rhopr-DH(31)-C, as determined using semi-quantitative Southern blot analysis. Fluorescent in situ hybridization reveals that Rhopr-DH(31) is expressed in a variety of cells in the CNS, including some neurosecretory cells.
Zandawala
Meet
M
Department of Biology, University of Toronto Mississauga, Mississauga, Ontario, Canada L5L 1C6. meet.zandawala@utoronto.ca
Paluzzi
Jean-Paul
JP
Orchard
Ian
I
D017398:Alternative Splicing / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D017404:In Situ Hybridization, Fluorescence; D007301:Insect Hormones / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012239:Rhodnius / Q000235:genetics*; D016415:Sequence Alignment
D018076:DNA, Complementary; D007301:Insect Hormones; D010455:Peptides; D012333:RNA, Messenger