schistoflrfamide and proctolin

We have developed a semi-intact preparation-consisting of an isolated oviduct with abdominal ganglia VII and VIII intact and attached-with which to characterize the effects on oviduct contraction, of peptides that are bath applied to CNS tissues. The work presented here offers a qualitative analysis of the central effects of SchistoFLRFamide and proctolin upon action potentials recorded from the oviducal nerves and upon oviduct contraction. In the process of this, we hope to demonstrate that a previously characterized putative CNS SchistoFLRFamide receptor [Peptides 23 (2002) 765] is a functional receptor.SchistoFLRFamide (10(-6)M), bath applied to abdominal ganglion VII, caused an increase in action potential frequencies recorded from the oviducal nerves, as well as an increase in the frequency of phasic contractions of the oviduct. Although the function of this response is not known, these results further support the possibility that the putative CNS SchistoFLRFamide receptors are functional receptors. Proctolin (10(-6)M), bath applied to abdominal ganglion VIII, altered the rhythmic bursting of action potentials recorded from the oviducal nerve and changed the appearance and cycle duration of neurogenic oviduct contractions.

Topics: Action Potentials; Animals; Female; Ganglia; Grasshoppers; Muscle Contraction; Neuropeptides; Oligopeptides; Oviducts

The midgut of insects has recently been shown to contain numerous endocrine-like cells and the midgut is now considered one of the largest endocrine organs in the insect. Using immunohistochemistry, radioimmunoassay, and muscle bioassay techniques, the midgut of the adult locust, Locusta migratoria, has been investigated for the distribution and possible function of FMRFamide-related peptides contained within these endocrine-like cells and innervation. Endocrine-like cells containing RFamide-like immunoreactivity were observed to be unequally distributed throughout the midgut. RFamide-like immunoreactivity was also seen in the ingluvial ganglion and in the nerves projecting posteriorly to the midgut. These axonal tracts resulted in extensive arborizations over the posterior midgut which were RFamide-like immunoreactive. Radioimmunoassay indicated larger amounts of FMRFamide equivalents in female locust midgut as compared to males with an unequal distribution of FMRFamide-like immunoreactivity in the gastric caeca and in the anterior and posterior parts of the midgut. Circular muscle contraction of the midgut was monitored using a ring-type preparation. Structure/activity studies have shown that the only FMRFamide-related peptides tested that alter circular muscle contraction of the midgut are those that belong to the subfamily referred to as myosuppressins. SchistoFLRFamide, leucomyosuppressin, and ManducaFLRFamide were each capable of lowering basal tonus and inhibiting spontaneous and proctolin-induced contractions of midgut muscle. Further structure/activity studies indicated that HVFLRFamide is the minimum sequence required to achieve inhibition comparable to the parent compound. This work suggests that a possible function for the FMRFamide-related peptides contained within the endocrine cells and innervation of the midgut of the locust may be in modulating midgut contraction and thereby playing a role in digestion.

Topics: Animals; Dose-Response Relationship, Drug; Female; Gastrointestinal Motility; Grasshoppers; In Vitro Techniques; Intestines; Male; Muscle Contraction; Neuropeptides; Oligopeptides; Structure-Activity Relationship

We have previously shown differential expression of leucomyosuppressin (LMS) mRNA in apparent endocrine cells in the anterior region of midguts of the cockroach Diploptera punctata, using in situ hybridization. In contrast, other FMRFamide-related peptides, as revealed by immunohistochemistry, have been found most abundantly in the posterior region in both apparent endocrine cells and nerve tracts. Here, we partially purified extracts of anterior and posterior cockroach midguts, using HPLC coupled with radioimmunoassay, and found, among multiple FMRFamide-like immunoreactive fractions, one fraction co-eluting with LMS in both regions. The presence of a co-eluting fraction in the posterior region, in the absence of LMS mRNA positive endocrine cells suggests that LMS might therefore be present in nerve tracts running along the length of the midgut. Using a circular muscle contraction assay from different portions of midgut, we determined the effects of LMS, proctolin and a variety of other midgut peptides on contractions of the midgut of Diploptera. Proctolin caused a sustained tonic contraction in the anterior midgut, the amplitude of which was dose-dependent. In contrast, LMS, and its relative SchistoFLRFamide, reduced the amplitude of these contractions. LMS and SchistoFLRFamide also inhibited spontaneous phasic contractions, which were elicited by proctolin application in only a few preparations. Other postulated midgut peptides did not induce or inhibit contractions, nor augment the proctolin-induced contractions. The C-terminal truncated sequences of LMS, HVFLRFamide and VFLRFamide, were sufficient to reduce the amplitude of the proctolin-induced contractions. This work illustrates a possible physiological role for LMS in Diploptera midguts, in the passage of food along the alimentary canal.

Topics: Animals; Cockroaches; FMRFamide; Muscle Contraction; Nerve Tissue Proteins; Neuropeptides; Oligopeptides; Peptide Fragments; Peptides