Isolation, cloning, and expression mapping of a gene encoding an antidiuretic hormone and other CAPA-related peptides in the disease vector, Rhodnius prolixus.
After a blood meal, Rhodnius prolixus undergoes a rapid diuresis to eliminate excess water and salts. During the voiding of this primary urine, R. prolixus acts as a vector of Chagas' disease, with the causative agent, Trypanosoma cruzi, infecting the human host via the urine. Diuresis in R. prolixus is under the neurohormonal control of serotonin and peptidergic diuretic hormones, and thus, diuretic hormones play an important role in the transmission of Chagas' disease. Although diuretic hormones may be degraded or excreted, resulting in the termination of diuresis, it would also seem appropriate, given the high rates of secretion, that a potent antidiuretic factor could be present and act to prevent excessive loss of water and salts after the postgorging diuresis. Despite the medical importance of R. prolixus, no genes for any neuropeptides have been cloned, including obviously, those that control diuresis. Here, using molecular biology in combination with matrix-assisted laser desorption ionization-time of flight-tandem mass spectrometry, we determined the sequence of the CAPA gene and CAPA-related peptides in R. prolixus, which includes a peptide with anti-diuretic activity. We have characterized the expression of mRNA encoding these peptides in various developmental stage and also examined the tissue-specific distribution in fifth-instars. The expression is localized to numerous bilaterally paired cell bodies within the central nervous system. In addition, our results show that RhoprCAPA gene expression is also associated with the testes, suggesting a novel role for this family of peptides in reproduction.
Paluzzi
Jean-Paul
JP
Department of Biology, University of Toronto Mississauga, South Building (Room 3016A), 3359 Mississauga Road North, Mississauga, Ontario, Canada L5L 1C6. jeanpaul.paluzzi@utoronto.ca
Russell
William K
WK
Nachman
Ronald J
RJ
Orchard
Ian
I
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D004199:Disease Vectors; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D012239:Rhodnius / Q000235:genetics* / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D014667:Vasopressins / Q000235:genetics* / Q000302:isolation & purification
D018076:DNA, Complementary; D029721:Drosophila Proteins; D009479:Neuropeptides; C455043:capa protein, Drosophila; D014667:Vasopressins