Effects of inhibitors and specific ion-free salines on segmental fluid secretion by the Malpighian tubules of the black field cricket Teleogryllus oceanicus

The effects of inhibitors and specific ion-free salines on fluid secretion rates in the distal and main segments showed that there were major differences in secretory mechanisms in the two segments. Both main and distal segments of the Malpighian tubules were sensitive to DIDs, SITS and acetazolamide but in different ways. The evidence suggests that the main segment does not contain a Cl(-)/HCO(3)(-) exchanger in the basal membrane, whereas the distal segment may do so. Secretion in both segments was K(+) dependent. Ba(2+) markedly reduced fluid secretion by the main segment and K(+) entry into the cells of the main segment is suggested to be predominantly via K(+) channels. Entry of K(+) may be primarily by other routes, such as Na K ATPase, in the distal segment. In the distal segment secretion was highly Mg(2+) dependent. Both segments were sensitive to amiloride analogs suggesting the presence of apical cation/H(+) exchangers.     

Control of ion and fluid transport by putative second messengers in different segments of the Malpighian tubules of the black field cricket Teleogryllus oceanicus

The differences in second messenger control of secretion were investigated in the distal and main segments of the Malpighian tubules of the black field cricket Teleogryllus oceanicus. Secretion by the main segment was considerably increased by corpora cardiaca extract and db-cAMP. Corpora cardiaca had no effect on secreted fluid composition or intracellular elemental composition but db-cAMP increased Na(+) and Cl(-) transport, as measured by x-ray microanalysis of secreted fluids and cells. Secretion by the main segment was considerably increased by forskolin and by Sp-cAMP. Secretion in the distal segment was abolished by corpora cardiaca extract but was unaffected by db-cAMP and only slightly reduced by 8-bromo-cAMP. However, Sp-cAMP increased secretion but forskolin reduced secretion. The responses of the distal segment suggest the possibility of a multiplicity of controls through different protein kinases and adenylyl cyclases. Secretion rate in the main segment was also increased by cGMP but distal segment secretion was unaffected. Secretion from both segments was increased by 5-HT. In the main segment secretion rate was increased by Ca-ionophore and thapsigargin and decreased by verapamil. This suggests a role for Ca(2+) as a controlling second messenger. In the distal segment only Ca-ionophore had an effect on secretion rate, which was reduced. Secretion rates in both segments were decreased in Ca-free saline. In saline in which Sr(2+) replaced Ca(2+), secretion rate in the main segment was greatly increased whilst that of the distal segment was decreased, suggesting that Sr(2+) could substitute for Ca(2+) as a second messenger.     

X-ray microanalysis of the Malpighian tubules of the black field cricket Teleogryllus oceanicus: the roles of Na K ATPase and the Na K 2Cl cotransporter

Both main and distal segments of the Malpighian tubules were sensitive to ouabain and furosemide but in different ways. Oubain had no effect on secretion rate by the main segment but in the secreted fluid Na(+) concentration increased substantially whereas K(+) decreased. Similarly intracellular elemental Na concentration increased and K decreased. Furosemide decreased the secretion rate of the main segment by 80%. The Na(+) concentration in the secreted fluid increased markedly but K(+) was not affected. Intracellular elemental Na concentration also increased but K was unchanged. In the distal segments both ouabain and furosemide decreased secretion rate by 40% but although ouabain had no effect on the composition of the secreted fluid, furosemide caused a substantial reduction in the concentrations of Mg(2+) and Cl(-) and a substantial increase in Na(+) and K(+) concentrations. The evidence suggests that the main segment contains a Na K ATPase and possibly a Na K 2Cl cotransporter whereas the distal segment may contain a Na K ATPase and a furosemide sensitive Mg(2+) transporter. K(+) entry into the cells of the main segment may be partially effected by a Na K 2Cl cotransporter but may be primarily via Na K ATPase in the distal segment.     

Use of Rb(+) and Br(-) as tracers for investigating ion transport by X-ray microanalysis in the Malpighian tubules of the black field cricket Teleogryllus oceanicus

Substitution of Rb(+) for K(+) in the incubation saline for in vitro preparations of Malpighian tubules had little effect on tubule function. Secretion rates increased by 10% for whole tubules, 9% for distal segments and 10% for main segments. In the secreted fluids Rb(+) almost completely replaced K(+). Within the cells of the main segment of the tubules Rb replaced the majority of the intracellular K. Treatment by ouabain in Rb saline resulted in a considerable increase in intracellular Na and Cl concentrations but no change in Rb concentration. This suggests that Rb(+) did not enter the cell via Na K ATPase and that the latter was not directly involved in Rb(+) secretion and by inference K(+) secretion. Substitution of Br(-) for Cl(-) in the incubation saline resulted in a 30% reduction in secretion rate from the distal segments but only a 10% reduction for the main segment. Cl(-) was almost completely replaced by Br(-) in fluid from both main and distal segments. In cells of the main segment the intracellular concentration of Br(-) did not exceed 30mmol kg(-1) dry weight and the Cl(-) concentration was unchanged in the apical region of the cell and increased in the basal region. These data suggest that Br(-) was transported across the tubule epithelium by a paracellular route and that the basal cell membrane is relatively impermeable to Cl(-). By inference Cl(-) may also be transported by a paracellular route.     

Acoustically-orienting parasitoids in calling and silent males of the field cricket Teleogryllus oceanicus

Abstract.

• 1 On three Hawaiian Islands, the introduced Australasian field cricket Teleogryllus oceanicus Le Guillou (Orthoptera: Gryllidae) was found to be attacked by the phonotactic parasitoid tachinid fly, Ormia ochracea Bigot.
• 2 Noncalling males occurred with callers in all locations, but silent males were more heavily parasitized than callers.
• 3 Body size was unrelated to both calling status and the likelihood of harbouring parasitoid larvae.
• 4 An experiment examining the likelihood of calling in the laboratory by males collected as silent or calling individuals showed no difference between the two classes of males, after accounting for parasitoid levels; males harbouring larvae were less likely to call.

     

Sexual selection on cuticular hydrocarbons in the Australian field cricket, Teleogryllus oceanicus

Background  

Females in a wide range of taxa have been shown to base their choice of mates on pheromone signals. However, little research has focussed specifically on the form and intensity of selection that mate choice imposes on the pheromone signal. Using multivariate selection analysis, we characterise directly the form and intensity of sexual selection acting on cuticular hydrocarbons, chemical compounds widely used in the selection of mates in insects. Using the Australian field cricket Teleogryllus oceanicus as a model organism, we use three measures of male attractiveness to estimate fitness; mating success, the duration of courtship required to elicit copulation, and subsequent spermatophore attachment duration.     

Cuticular hydrocarbons are heritable in the cricket Teleogryllus oceanicus

The ability of individuals to respond differentially to conspecifics depending on their genetic relatedness is a widespread phenomenon across the animal kingdom. Despite this, little is known about the selection processes that act on the phenotypic variation of traits used during recognition. Here we use a quantitative genetic approach to examine the patterns of genetic variation in cuticular hydrocarbon (CHC) profiles, a pheromonal system used extensively in insect communication. Using gas chromatography, we found family specificity in the CHC profiles of male crickets, Teleogryllus oceanicus. Across CHC peaks, our mean coefficient of additive genetic variation was 10.8%. Multivariate principal component analysis showed that most axes of variation were weighted by CHC peaks with significant additive genetic variation. Our results provide evidence that variation in CHC profiles can reflect genetic relatedness, supporting the widely held belief that this phenotypic trait is used as a mechanism for chemosensory kin recognition.     

Geographic variation in female preference functions and male songs of the field cricket Teleogryllus oceanicus

Male crickets (Teleogryllus oceanicus) produce a complex call consisting of two elements, the long chirp (three to eight sound pulses) followed by a series of short chirps (each with two sound pulses). There is significant geographic variation in the temporal structure of calls, and the long chirp is selected against by acoustically orienting parasitoids in some populations. Here we examine geographic variation in female preference functions for the amount of long chirp. In general, females prefer calls with greater proportions of long chirp, although the strength and nature of selection varied across populations. Variation in preference functions did not match variation in call structure. There was a mismatch between the proportion of long chirp produced by males in a population and the proportion of long chirp preferred by females. The convergent preferences of predators and females are likely to maintain genetic variation in song traits in parasitized populations. The apparent mismatch between preference and trait is discussed in relation to theoretical models of preference evolution.     

Microsatellite loci for the Australian field cricket Teleogryllus oceanicus and their cross‐utility in Teleogryllus commodus

Microsatellite loci were isolated from the Australian field cricket Teleogryllus oceanicus. Seven polymorphic loci were found with an observed number of alleles ranging from eight to 17 and observed heterozygosities between 0.26 and 0.94. One locus was found to be X‐linked. These seven loci were also tested for amplification and polymorphism in the congeneric species Teleogryllus commodus. The loci will be used for paternity studies in these species.     

Preexisting behavior facilitated the loss of a sexual signal in the field cricket Teleogryllus oceanicus

Behavioral preadaptations can provide an accommodating environmentin which novel morphological characters may be selected. A veryrecent morphological mutation, flatwing, has caused the lossof male song in field crickets (Teleogryllus oceanicus) fromthe Hawaiian island of Kauai. Previous studies in this and relatedspecies have shown that females require male courtship songto mount males, but flatwing T. oceanicus males still achievematings. Females from Kauai may have a relaxed requirement formale courtship song, or flatwing males may compensate for theirinability to sing by altering other courtship behaviors. Wetested whether male courtship and female responses to male courtshipwere preadapted in a way that facilitated the spread of themale wing mutation or if parallel changes in male courtshipand female responses accompanied the mutation. We performedmating trials in 2 captive-bred populations to assess how matingbehavior varied depending on the presence or absence of courtshipsong playback. The first was an ancestral population from Kauaiestablished prior to the emergence of the flatwing mutation,and the second was derived from Kauai after the mutation becameprevalent. Mating behaviors did not differ qualitatively orquantitatively between the ancestral and current populations,and females from both accepted males for mating in the absenceof courtship song. Our results provide direct evidence thata mechanism allowing flatwing males to mate with females wasin place before the mutation actually arose on Kauai and demonstratehow preexisting behavior facilitated the rapid spread of a novelmorphological mutation.     

Auditory pattern recognition and steering in the cricket Teleogryllus oceanicus

Male crickets Teleogryllus oceanicus (Le Guillou) produce a complex species‐specific calling song with phrases combining groups of single pulses (chirps) and groups of double‐pulses (trills) to attract females, which fly or walk towards singing males. In open‐loop trackball experiments, phonotactic steering responses to normal calling song phrases consisting of chirps and trills are strongest, suggesting that both components are necessary for maximal attractiveness. Sequences of just chirps or trills are less effective in eliciting phonotactic walking and steering. Split‐song paradigms are used to analyze the steering behaviour underlying orientation in more detail. The females' phonotactic steering reflects the alternating acoustic pattern of the split‐song paradigm. Analysis with high temporal resolution demonstrate, that even when the calling song is presented only from one side, the steering velocity and lateral deviation towards the song is modulated by steering events to single‐sound pulses. Therefore, pattern recognition, which integrates the structure of the song, appears not to be directly involved in the rapid steering response. This organization of phonotactic behaviour with a parallel processing of pattern recognition and steering is similar to other cricket species and may allow T. oceanicus females to steer transiently towards distorted song patterns as they occur in natural habitats.     

Cuticular hydrocarbons influence female attractiveness to males in the Australian field cricket,Teleogryllus oceanicus

Sexual dimorphism is thought to result from directional sexual selection acting on male signal traits, with female signal traits given little, if any, attention. Here, we examine male mating preferences in the Australian field cricket, Teleogryllus oceanicus. Using a multivariate selection analysis approach, we found that male preferences have the potential to exert selection on female cuticular hydrocarbons, chemical compounds widely used as sexual signals in insects. In addition to finding both stabilizing and disruptive preference gradients, we also found weak negative directional preference for female cuticular hydrocarbons. We contrast our results with a recent study examining sexual selection via female choice on male T. oceanicus cuticular hydrocarbons and suggest that differences in the form and intensity of sexual selection between the genders may provide part of the net selection differential necessary for the evolution of sexual dimorphism in this species.     

Geographical variation in calling song of the field cricket Teleogryllus oceanicus: the importance of spatial scale

We compared calling songs of the field cricket Teleogryllus oceanicus from 15 sites within six regions of two continental areas, Oceania and Australia. The cricket was introduced to Hawaii, where it is subject to an acoustically orienting parasitoid fly not found elsewhere in its range. In a principal components analysis (PCA) of song from all populations combined, the first five components had eigenvalues greater than one, and collectively accounted for over 80% of the total variation. Means for all song components varied significantly among sites, and different components varied at the three levels of analysis (continent, region and site). The principal way in which sites differed was along a gradient in increasing song length, pulse duration and intervals between song elements. Crickets from Oceania had a significantly greater variance in their song than Australian crickets, driven largely by the high variance in Hawaii. Geography explained a substantial amount of variation in song, despite the likelihood of serial bottlenecks having occurred as the species moved from island to island. Because female crickets appear not to focus on the short chirp as a component of mate choice, a lack of selection may allow this song component to vary more widely.     

Stimulation of fluid secretion by single isolated Malpighian tubules of the house cricket, Acheta domesticus

Abstract. Fluid secretion by single isolated Malpighian tubules of Acheta domesticus (L.) is stimulated by aqueous extracts of nervous and neuroendocrine tissues from the cricket. Diuretic activity, expressed as the increase in rate of secretion per microgram tissue protein (pl/mm/min/ug protein), is highest in the CA and CC.
The response to aqueous CC extracts is dose-dependent. The maximum increase in secretion rate is 300–350 pl/mm/min and the ED_max and ED₅₀ are 0.1–0.2 and 0.02–0.03 gland pairs respectively.
The diuretic activity in the CC is retained after 5min at 95C, and is freely soluble in 80% methanol. Diuretic activity is, however, greatly reduced after prolonged heating or after treatment with either pronase or chymotrypsin. It is conluded that the diuretic factor(s) is a low molecular weight peptide. A loss of activity after incubation with pyroglutamate amino peptidase suggests that some active peptides present are N-terminally blocked.
The diuretic activity of crude aqueous CC extracts is rapidly lost on standing at room temperature. This is partly prevented by precipitation of protein enzymes by either heat treatment or extraction in methanol.
Synthetic vertebrate and insect neuropeptides are generally low in activity increasing fluid secretion by no more than 60pl/mm/min. However, extracts of neuroendocrine tissues from a wide range of insect species are potent stimulants of tubule secretion. Various biogenic amines were tested and have little effect on fluid secretion; thus they cannot contribute greatly to the response of cricket tubules to aqueous tissue extracts.     

Sexual dimorphism in cuticular hydrocarbons of the Australian field cricket Teleogryllus oceanicus (Orthoptera: Gryllidae)

Sexual dimorphism is presumed to reflect adaptive divergence in response to selection favouring different optimal character states in the two sexes. Here, we analyse patterns of sexual dimorphism in the cuticular hydrocarbons of the Australian field cricket Teleogryllus oceanicus using gas chromatography. Ten of the 25 peaks found in our chromatographs, differed in their relative abundance between the sexes. The presence of sexual dimorphism in T. oceanicus is discussed in reference to a review of sexual dimorphism in cuticular hydrocarbons of other insects. We found that this trait has been examined in 103 species across seven different orders. Seventy-six of these species (73%) displayed sex specificity of cuticular hydrocarbons, the presence/absence of which does not appear to be directly linked to phylogeny. The occurrence of sexual dimorphism in cuticular hydrocarbons of some but not other species, and the extent of variation within genera, suggest that this divergence has been driven primarily by sexual selection.     

Regulation of haemolymph sodium and potassium during dehydration in the cricket Teleogryllus oceanicus

ABSTRACT. Male crickets ( Teleogryllus oceanicus ) when dehydrated for 3 days lost 51% of their body water, and 65% of their haemolymph volume. Haemolymph osmolality rose from 391 to 572mOs/kg; [Na⁺] from 149 to 289 HIM; and [K⁺] from 13.0 to 26.3 mM. During dehydration 385 μig Na (expressed as NaCl) and 41 μug K (expressed as KCI) were removed from the haemolymph. Rehydration of the dehydrated insects failed to restore the Na⁺ and K⁺ concentrations to near their original levels. Approximately 62% of the missing Na⁺ was excreted, whilst five times the amount of K⁺ removed from the haemolymph was excreted. It is presumed that the excess represents K⁺ removed from intracellular fluid.     

Sexual signaling and immune function in the black field cricket Teleogryllus commodus

The immunocompetence handicap hypothesis predicts that male sexual trait expression should be positively correlated with immunocompetence. Here we investigate if immune function in the cricket, Teleogryllus commodus, is related to specific individual components of male sexual signals, as well as to certain multivariate combinations of these components that females most strongly prefer. Male T. commodus produce both advertisement and courtship calls prior to mating. We measured fine-scale structural parameters of both call types and also recorded nightly advertisement calling effort. We then measured two standard indices of immune function: lysozyme-like activity of the haemolymph and haemocyte counts. We found a weak, positive relationship between advertisement calling effort and lysozyme-like activity. There was, however, little evidence that individual structural call components or the net multivariate attractiveness of either call type signalled immune function. The relationships between immunity and sexual signaling did not differ between inbred and outbred males. Our data suggest that it is unlikely that females assess overall male immune function using male calls.     

Serotonin has kinin-like activity in stimulating secretion by Malpighian tubules of the house cricket Acheta domesticus

Serotonin stimulates secretion by Malpighian tubules (MT) of a number of insects, and functions as a diuretic hormone in Rhodnius prolixus and in larval Aedes aegypti. Serotonin is here shown to be a potent stimulant of secretion by MT of the house cricket, Acheta domesticus, with an apparent EC₅₀ of 9.4 nmol L⁻¹, although its diuretic activity is just 25% of the maximum achievable with either the native CRF-related peptide, Achdo-DH, or a crude extract of the corpora cardiaca. In this respect, the diuretic activity of serotonin is similar to that of the cricket kinin Achdo-KI, and when tested together their actions are not additive, which suggests they target the same transport process. Consistent with this suggestion, the activity of serotonin is chloride-dependent and is associated with a non-selective stimulation of NaCl and KCl transport. In common with Achdo-KI, serotonin has no effect on cAMP production by isolated MT, and both act synergistically with exogenous 8bromo-cAMP in stimulating fluid secretion, most likely by promoting the release of Ca²⁺ from intracellular stores. A number of serotonin agonists and antagonists were tested to determine the pharmacological profile of receptors on cricket MT. The results are consistent with the diuretic activity of serotonin being mediated through a 5-HT₂-like receptor.