Biological activity of diuretic factors on the anterior midgut of the blood-feeding bug, Rhodnius prolixus

Probing of a host and ingestion of a blood-meal in a fifth instar Rhodnius prolixus results in a cascade of tightly integrated events, including salivary gland secretion, plasticisation of the abdominal cuticle, increased ion and water movement across the anterior midgut (crop) and Malpighian tubules (which rapidly produce urine) and the regular expulsion of urine from the hindgut. In this study we have focussed on the role of the anterior midgut during the rapid postprandial diuresis. The huge blood-meal is pumped into the anterior midgut, during feeding, then modified by diuresis and stored until it is digested. Changes in the anterior midgut activity are rapid. Within minutes of the commencement of feeding there is an increase in the frequency of anterior midgut contractions and diuresis begins with the movement of salt and water across the epithelium of the anterior midgut into the haemolymph.While serotonin, a diuretic hormone in R. prolixus, is known to play a role in the physiological activity of the anterior midgut, we were interested in exploring further the role of serotonin, and other diuretic peptides. We have tested the activity of several peptides, including R. prolixus calcitonin-like diuretic hormone (Rhopr-DH31), corticotropin-releasing factor (CRF)-like peptide from Zootermopsis nevadensis DH (Zoone-DH) and a kinin from Leucophaea maderae, Leucokinin 1 (LK1). These peptides families are known to be present in the central nervous system of R. prolixus, are putative neurohormones released into the haemolymph after the start of feeding, and have been shown to have activity on a variety of tissues involved in post-feeding diuresis. We show here that both serotonin and Zoone-DH increase the cAMP content of the anterior midgut and that serotonin, Zoone-DH and cAMP analogues increase absorption of water from the anterior midgut, increase the short circuit current and voltage, while decreasing the resistance across the epithelium. While LK1 and Rhopr-DH31 do not significantly increase absorption, or short circuit current, LK1 does significantly decrease the resistance and transepithelial voltage of the anterior midgut epithelium. All of the factors studied increase the frequency of contractions of the anterior midgut.