Length heterogeneity of amplified circular rDNA molecules in oocytes of the house cricket Acheta domesticus (Orthoptera: Gryllidae)

Amplification of the genes coding for rRNA occurs in the oocytes of a wide variety of organisms. The amplification process appears to be mediated through a rolling-circle mechanism. The approximate molecular weight of the smallest rDNA circles is equivalent to the estimated combined molecular weight of DNA which codes for a single ribosomal RNA precursor molecule and an associated non-transcribed spacer DNA sequence. RNA-DNA hybridization studies carried out on oocytes of the house cricket, Acheta domesticus, suggest that DNA coding for rRNA accounts for only a small fraction of the rDNA satellite, all of which is amplified in the oocyte. In order to test the possibility that the remainder of the amplified rDNA represents spacer and to determine whether a rolling-circle mechanism might also be involved in amplification in A. domesticus oocytes, rDNA was isolated from ovaries of A. domesticus and spread for electron microscopy. A large proportion of the rDNA isolated from ovaries is circular, while main-band DNA and rDNA prepared from other tissues demonstrates few if any circles. The mean size of the smallest rDNA circles is approximately 8 times longer than the length estimated for DNA which codes for 18 S and 28 S rRNA. Denaturation mapping shows the rDNA circles to contain two major readily denaturing regions located about equidistant from one another on the circle. Each readily denaturing region accounts for 4–6% of the total DNA in the circle. The fact that only 12% of the average molecule is required to code for A. domesticus 18 S and 28 S rRNA is consistent with the hybridization data. Considerable size heterogeneity exists in the length of the smallest class of rDNA molecules. In the rDNA of other species such heterogeneity has been shown to reside in the non-transcribed spacer.     

Tracheation of abdominal ganglia and cerci in the house cricket Acheta domesticus (Orthoptera,Gryllidae)

Patterns of tracheation in the abdominal central nervous system and the cerci of Acheta domesticus are described from whole mounts, and light and electron microscopy. The tracheal supply of the ganglia is derived from ventral longitudinal tracheal trunks which have segmental connections to the spiracels. Each abdominal ganglion is served by a single pair of tracheal trunks, except the terminal ganglion, which has two pairs. Within the ganglia, tracheoles occur principally in association with glia-rich areas of the neuropile. We suggest that the respiratory exchange may be concentrated in the cell bodies of neurons and glia. Each cercus has a tracheal supply in paralle with a large air sac which, it is suggested, serves to lighten the cercus, functions as a resonator for sound reception, or facilitates tidal flow of hemolymph and postecdysial expansion of the cercus. No tracheae run continuously between ganglia or between the terminal ganglion and the cerci, and they do not appear to have a potential role as a contact guidance pathway for cercal nerve growth.     

Analysis of a large nontranscribed spacer in the ribosomal DNA of the house cricket,Acheta domesticus (Orthoptera:Gryllidae)

An analysis of a 29-kilobase nontranscribed spacer fragment in the ribosomal DNA (rDNA) of the house cricket, Acheta domesticus, revealed a highly repetitious structure. A total of eight EcoRI repeats of three different size classes measuring 259, 420, and 508 base pairs (bp) was mapped to a region 2 kilobases (kb) from the 18 S coding region. The repeats were oriented in a nonrandom manner and had sequences homologous to DNA located immediately adjacent to the repetitive array. DNA sequence analysis showed that the repetitive region was composed of smaller direct repeats 66, 67, and 383 bp in length. There was minor length heterogeneity of the chromosomal restriction fragments containing the entire array, indicating that a variable number of EcoRI repeats is a minor contributor to the total repeat-unit length heterogeneity. Immediately upstream from the EcoRI array there is a 17-kb region composed of 50 to 60 subrepeat elements recognized by a variety of restriction endonucleases. A subcloned SmaI repeat from the array was not homologous to any other part of the rDNA repeat unit or other chromosomal DNA. There was little length heterogeneity in restriction fragments containing the chromosomal 17-kb repetitions region. Immediately upstream from the 17-Kb region there is a 4.1-kb segment with sequences homologous to the EcoRI repeats.     

The delta 12-desaturase from the house cricket, Acheta domesticus (Orthoptera: Gryllidae): characterization and form of the substrate

A novel delta 12-desaturase from animals, which converts oleic acid (18:1n-9) to linoleic acid (18:2n-6), was characterized in the house cricket, Acheta domesticus. The delta 12-desaturase product, linoleic acid, was determined by silver nitrate thin-layer chromatography, radio-gas-liquid chromatography and radio-high-performance liquid chromatography with the latter being used for routine analyses. Enzyme activity was located in the microsomal fraction of whole insect homogenates. NADPH or NADH was required for activity, with NADPH being the more efficient electron donor. In short incubation times with oleoyl-CoA as substrate, the highest amount of product, linoleic acid, was found as linoleoyl-CoA. With longer incubation periods, most of the linoleic acid was recovered in the polar lipid fraction containing phospholipid. Preincubation of the microsomal preparation in the absence of NADPH, which allowed 90% of the oleoyl moiety to be transacylated into complex lipid, resulted in no detectable desaturation upon addition of NADPH. These data indicate that the oleic acid moiety used as substrate was in the form of a CoA derivative and not in the form of a phospholipid, as it is for the plant delta 12-desaturase. This is the first characterization of a delta 12-desaturase from an animal system and the first report of a delta 12-desaturase that uses oleoyl-CoA as substrate.     

Courtship song, male agonistic encounters, and female mate choice in the house cricket,Acheta domesticus (Orthoptera: Gryllidae)

Whether female crickets choose among males based on characteristics of the courtship song is uncertain, but in many species, males not producing courtship song do not mate. In the house cricket,Acheta domesticus, we examined whether a female chose or rejected a male based on his size, latency to chirp, latency to produce courtship song, or rate of the high-frequency pulse of courtship song (“court rate”). We confirmed that females mated only with males that produced courtship song, but we found no evidence that the other factors we measured affected a female’s decision to mate. In addition, we investigated whether the outcome of male agonistic encounters affected the subsequent production of courtship song. In one experiment, we observed courtship and mating behavior when a single female was placed with a pair of males following a 10-min interaction period between the two males. Winners of male agonistic encounters had higher mating success. However, winners and losers of agonistic encounters were not different in their likelihood or latency to produce courtship song or in the number of times they were disrupted by the other male in the pair. In a second experiment, we allowed two males to interact for a 10-min period, but following this interaction period, we placed a female with each male separately and observed courtship and mating behavior. The mating success of winners and losers was not different under these circumstances, and we found no differences between winners and losers in any subsequent courtship or mating behavior examined. We conclude that winning agonistic encounters influences a male’s mating success in ways other than his production of courtship song and this effect is lost when winning and losing males are separated and each is given an opportunity to mate.     

Male mate choice based on chemical cues in the cricket Acheta domesticus (Orthoptera: Gryllidae)

1. Males in many animal species exercise mate choice to maximise their reproductive success, assessing females by characteristics related to reproductive potential, such as mating status, body size, and age. The sensory modalities involved in mate choice are often not firmly demonstrated, but only inferred. This is especially true for chemical cues and signals. 2. The present study tests whether males of the cricket Acheta domesticus are able to choose among females based only on chemosensory cues. In A. domesticus, as in many crickets, males call to attract females or roam the habitat silently to search for females. In three‐way choice trials, males were presented with two filter papers that had been placed with females for 24 h prior to the trials and one blank control. Females were either mated or virgin and starved or well‐fed. It was predicted that males would prefer virgin over mated females and those in good condition over starved ones. 3. Males were more likely to contact filters that had been exposed to females. They spent more time examining filter papers from virgin females than those from mated ones, while the condition of the females had no effect. 4. We conclude that males can detect chemical cues from females on substrate and distinguish virgin females from mated ones. Being able to assess sperm competition risk prior to mating or even before further pursuing a trail with chemical cues should confer a considerable benefit to males.     

Cajal bodies (coiled bodies) in the nuclei of the house cricket (Acheta domesticus) oocytes

The morphology and fine structure of Cajal bodies (coiled bodies, CB) in the germinal vesicles (oocyte nuclei) of the house cricket, Acheta domesticus have been analyzed. It is shown that in the studied species CBs arise as early as in the youngest previtellogenic oocytes, and are located next to or within aggregations of multiple nucleoli. Surprisingly, two morphological types of CBs have been found in the analyzed specimens. On the basis of EM studies we suggest that they represent subsequent developmental stages of CB morphogenesis.     

Temperature and the energetics of development in the house cricket (Acheta domesticus)

The influence of rearing temperature on the energetics of development was investigated in house crickets (Acheta domesticus). Crickets raised at 25 degrees C grew slower (0.51 mg d(-1), dry mass basis) and took longer to develop (119 d) but obtained a greater adult body mass (61 mg, dry mass) than crickets reared at 28 degrees C (0.99 mg d(-1), 49 d, 48 mg). Total metabolic energy consumed during development at 25 degrees C (1351 J) was twice that at 28 degrees C (580 J) primarily because of the longer development period, and as a consequence the specific net cost of growth was much greater for crickets reared at 25 degrees C (22.1 kJ g(-1)) than 28 degrees C (11.9 kJ g(-1)).     

Postembryonic development of teratogen-induced supernumerary eyes in Acheta domesticus (L.) (Orthoptera : Gryllidae)

Supernumerary compound eyes were generated in the cricket Acheta domesticus (L.) (Orthoptera : Gryllidae) by applying the insect teratogen biquidone (benz[g]isoquinoline-5, 10-dione) and partially characterized teratogenic mixtures to eggs within 24 hr following oviposition at 31 ± 0.5°C. Postembryonic development of supernumerary eyes was compared with that of normal compound eyes, using light and scanning electron microscopy. All elements of the dioptric apparatus were present in supernumerary eyes of nymphs, but no axonal connection to the brain was found. Supernumerary eyes deteriorated during successive instars. Occasionally, these eyes were sloughed off before the adult stage was reached; more frequently, eyes appeared as highly pigmented protrusions not readily identifiable as adult eye tissue. Teratogen-induced supernumerary eyes are probably not functional light receptors in A. domesticus nymphs or adults.     

Excretion in the house cricket (Acheta domesticus): ultrastructure of the ampulla and ureter

An ultrastructural analysis of the ampulla and ureter of the cricket, Acheta domesticus, is presented. The excretory system of the cricket is unusual in that the 112 Malpighian tubules do not attach directly to the gut, but fuse to form a bladder-like ampulla which is joined to the colon by a muscular ureter. The ampulla consists of two cell types, primary and regenerative. Primary cells secrete large numbers of membrane-bound vesicles into the lumen and also appear to be involved in fluid reabsorption. Regenerative cells are very small and form a layer just beneath the basal lamina of the ampulla. They are believed to differentiate and replace sloughed off primary cells. The ureter is a muscular tube lined with cuticle which connects the ampulla (endoderm) with the colon (ectoderm). The probable origin and significance of the morphological modifications of the excretory system are discussed.     

Excretion in the house cricket (Acheta domesticus): fine structure of the Malpighian tubules

An ultrastructural analysis of the Malpighian tubules of the cricket, Acheta domesticus, is presented. The excretory system of the cricket is unusual in that the 112 Malpighian tubules do not attach directly to the gut, but fuse to form a bladder-like ampulla which is joined to the colon by a muscular ureter. The tubules have three structurally distinct segments and consist of four cell types. Attached by basal lamina to the outer surface of the distal tip are nodules, consisting of small cells on membranous stalks. These are presumed to serve as attachments to the body wall. The distal 20% of the tubule is hyaline, consisting of a monolayer of squamous cells that appear to be secretory. The mid-tubule comprises 75% of the total length and is the primary region for fluid secretion. It is also characterized by having large numbers of laminate spheres in the cytoplasm of the cells. The proximal 5% of each tubule consists of more columnar cells and may function in fluid resorption. The relationship between structural features and known physiological functions are discussed.     

Fatty acid and lipid analysis of the house cricket,Acheta domesticus

The fatty acid content and composition of the house cricket Acheta domesticus have been investigated in entire insects at different developmental stages and in selected organs of male and female adults. We have also determined the fatty acid composition of the various lipid classes within extracts of the organs of adult female insects. Fatty acids were analysed by capillary gas chromatography or mass spectrometry as their methyl esters (FAMEs) after direct transesterification of insect material or separated lipid classes.The major esterified fatty acids in all extracts were palmitate (C_16:0), stearate (C_18:0), oleate (C_18:1) and linoleate (C_18:2). Levels of esterified fatty acid varied considerably between organs but the fatty acid compositions showed only small variations. The levels of polyunsaturated fatty acids of the C₁₈ series were considerably higher in phospholipid fractions than in other lipid classes. Triacylglycerols formed the major lipid class in ovaries, fat-body and newly-laid eggs, whereas diacylglycerols and phospholipid predominate in the haemolymph. Triacylglycerols, phospholipids, diacylglycerols and free fatty acids were all found in significant amounts in the gut tissue.     

The maltase of the house cricket Acheta domesticus L. (orthoptera: gryllidae)

• 1.1. The optimal pH of maltase of the house cricket was 5.6 and the optimal temperature was 40°C. The apparent activation energy was 5300 cal/mol
• 2.2. The 50% inactivation time for maltase at 50°C was 69 min and the 50% inhibition concentration for mercuric chloride was 163μmole.
• 3.3. Maltase was only produced by the caecal tissues but its highest activity was found in the foregut lumen and after 5 days of starvation, only 44.6% of the total maltase activity remained.

     

Nanomechanical properties of wing membrane layers in the house cricket (Acheta domesticus Linnaeus)

Many insect wings change shape dynamically during the wingbeat cycle, and these deformations have the potential to confer energetic and aerodynamic benefits during flight. Due to the lack of musculature within the wing itself, the changing form of the wing is determined primarily by its passive response to inertial and aerodynamic forces. This response is in part controlled by the wing’s mechanical properties, which vary across the membrane to produce regions of differing stiffness. Previous studies of wing mechanical properties have largely focused on surface or bulk measurements, but this ignores the layered nature of the wing. In our work, we investigated the mechanical properties of the wings of the house cricket (Acheta domesticus) with the aim of determining differences between layers within the wing. Nanoindentation was performed on both the surface and the interior layers of cross-sectioned samples of the wing to measure the Young’s modulus and hardness of the outer- and innermost layers. The results demonstrate that the interior of the wing is stiffer than the surface, and both properties vary across the wing.     

The energetic costs of fighting in the house cricket, Acheta domesticus L

The cost of performing an agonistic behavior, or tactic, willhave consequences for an individual's rate of cost accrual,the tactic's evolutionary stability if used as an assessmentsignal, and its pattern of use in the behavioral choreographyof a contest. Few studies have attempted to quantify the costsof fighting, particularly with regard to energy expenditure.Flow-through respirometry revealed that house cricket malescan expend energy at relatively high rates when fighting (morethan eight times resting levels) depending on the particulartactic performed. Acoustic signalling (stridulation) constitutedthe least costly of seven measured agonistic tactics, whilewrestling with an opponent, the most energetically costly tactic,consumed oxygen at a net rate more than 40 times that of stridulation.Low-cost tactics are used by opponents more frequently thanmore costly tactics, providing evidence for an escalating tacticalconvention based on energetic costs. The moderate energeticcost of some tactics suggests they may function as reliablesignals in the assessment of wrestling ability. Net energy costsper contest increased linearly with contest duration for bothcontest winners and losers, but winners tended to expend moreenergy per contest than losers. The likely fitness effects ofenergy expended while fighting are discussed. The results ofthis study indicate that energy expenditure is an importantcost shaping contest strategies in Acheta domesticus