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Enkephalins play a major role in reproductive physiology in crustaceans; however their role in reproductive development in insects is largely unknown. We investigated the effect of exposure to exogenous leucine-enkephalin (Leu-Enk), methionine-enkephalin (Met-Enk), and the opioid antagonist naloxone on gonad development in the Eastern lubber grasshopper, Romalea microptera. Injection of either Leu-Enk or naloxone alone significantly increased the testicular index and testicular follicular diameter in males, and the ovarian index, oocyte length, and oocyte diameter in females. In contrast, injection of Met-Enk inhibited all measures of reproductive development in both sexes. Surprisingly, co-injection of naloxone with either enkephalin enhanced the effect associated with administration of the enkephalin alone. This study clearly demonstrates the ability of enkephalins to disrupt insect sexual development and also suggests the existence of conserved enkephaline-dependent regulatory mechanisms in insects and crustaceans.

The interaction of juvenile hormone (JH) and nutrition was studied during the oviposition cycle of the Eastern Lubber grasshopper (Romalea microptera). Starvation of females early or in the middle of the cycle inhibited oocyte growth. Starvation for 4 days also reduced hemolymph levels of JH III and vitellogenesis (Vg) to 25% and 15%, respectively, of the levels in fed animals. Likewise, Vg production by fat body fragments incubated in vitro was reduced to 2% of the levels in fed animals and total protein synthesis was reduced to 25%, suggesting that starvation had a stronger effect on Vg synthesis than on protein synthesis. These effects were reversed when starved animals were fed again. However, fat body levels of Vg-mRNA were similar in fed and starved animals, indicating that starvation did not affect transcript levels. We tested whether the decline in JH levels mediated the other starvation effects by infusing animals with JH III or vehicle for 2 days at the onset of starvation. Infusion of JH elevated JH and Vg-mRNA levels 670% and 103%, respectively, above the levels in vehicle-infused animals. However, Vg production and hemolymph levels of Vg were similar to the levels in vehicle-infused animals. These data suggest that JH alone is insufficient to stimulate Vg production.

The in vitro synthesis of juvenile hormone (JH) by corpora allata (CA) from the lubber grasshopper (Romalea microptera) was stimulated by low concentrations of brain extract and this effect was reduced at higher concentrations, suggesting the presence of allatotropin (AT) and allatostatin (AST) factors in the brain. The AT activity of brain extracts caused a rapid and reversible stimulation and appeared to be a peptide(s). Reversed phase (C18) HPLC analysis of brain extracts disclosed two peaks of AT activity but no significant AST activity. Manse-AT, Schgr-NPF, and Locmi-FLRF had no effect on JH synthesis by lubber CA, indicating that the Rommi-AT factors are distinct from these peptides. High concentrations of Dippu-AST-7 and Grybi-AST-1 inhibited JH synthesis, implying that AST factors might be present in lubber grasshoppers. CA response to AT activity of brain extracts varied during the oviposition cycle ( approximately 35 days), with the maximum response occurring on days 16-18. AT activity of brain extracts also varied during the cycle, being highest on day 25. Our data suggest that the lubber CA is largely regulated by AT activity, and that JH synthesis reflects both CA response to AT activity and the level of AT activity in the brain.

Although, in many insects, migration imposes a cost in terms of timing or amount of reproduction, in the migratory grasshopper Melanoplus sanguinipes performance of long-duration flight to voluntary cessation or exhaustion accelerates the onset of first reproduction and enhances reproductive success over the entire lifetime of the insect. Since juvenile hormone (JH) is involved in the control of reproduction in most species, we examined JH titer after long flight using a chiral selective radioimmunoassay. JH levels increased on days 5 and 8 in animals flown to exhaustion on day 4 but not in 1-h or non-flier controls. No difference was seen in the diel pattern of JH titer, but hemolymph samples were taken between 5 and 7 h after lights on. Treatment of grasshoppers with JH-III mimicked the effect of long-duration flight in the induction of early reproduction. The increased JH titer induced by performance of long-duration flight is thus at least one component of flight-enhanced reproduction. To test the possibility that post-flight JH titer increases are caused by adipokinetic hormone (AKH) released during long flights, a series of injections of physiological doses of Lom-AKH I were given to unflown animals to simulate AKH release during long flight. This treatment had no effect on JH titers. Thus, although AKH is released during flight and controls lipid mobilization, it is not the factor responsible for increased JH titers after long-duration flight.

The role of hemolymph ecdysteroids in the reproduction of non-dipteran insects is unclear. We examine the role(s) of hemolymph ecdysteroids during egg production in the lubber grasshopper, Romalea microptera. In all individuals, hemolymph ecdysteroids rose to a sharp peak with similar maxima and then fell to undetectable levels. The time from the adult molt to the maximum ecdysteroid titer (E(max) titer) varied in response to food availability, whereas the time from E(max) titer to oviposition was unrelated to food availability. Because both the timing of egg production and the timing of E(max) responded similarly to environmental changes, ecdysteroids may be involved in egg production. We hypothesized that this role is the stimulation of vitellogenesis. Ovariectomized females had vitellogenin but no ecdysteroids, so ecdysteroids are not necessary for vitellogenin production. In addition, treatment of females with ecdysteroids altered neither Vg titers nor ovarian growth. Ovarian ecdysteriods increased at the same age in development as hemolymph ecdysteroids. In contrast to hemolymph ecdysteroids, ovarian ecdysteroids persisted until oviposition. Despite this, [(3)H]ecdysone injected into the hemolymph was detected later only at very low levels in the ovary, suggesting that hemolymph ecdysteroids are not sequestered by the ovary. In summary, our studies indicate that hemolymph ecdysteroids in adult females of the lubber grasshopper are associated with the timing of egg production, but they neither regulate vitellogenesis nor act as a source of ecdysteroids for the ovary., (Copyright 2003 Wiley-Liss, Inc.)

The genetic and geographical variation, gene flow, and the historical biogeography of the eastern lubber grasshopper, Romalea microptera (=guttata) (Houttuyn), were examined by sequencing a 420-bp region of the mitochondrial cytochrome b gene. Individuals (168) were collected from 12 sites in the southern United States that covered most of the range reported for this species. These populations contained 49 mitochondrial DNA haplotypes. A high level of genetic diversity was observed in these populations (3.8%), most of which was due to variation within populations. The highest genetic variation was found in a northern Florida population (collected at St. Marks, FL), and the lowest was found in a southern Florida population (Copeland, FL). Estimates of historical and current population sizes suggest that most of the lubber populations drastically declined in size at some point in the past. In contrast to previous studies on several other species in this region, phylogenetic analyses (PAUP) and haplotype age phylogenies (PHYLIP) showed no major geographic structure. These observations suggest that the distribution of this species in the past may have been homogeneous, rather than the patchy distribution that is currently observed. Alternatively, this result may reflect the absence of long-term barriers for the dispersal of this species. Either of these might have contributed to the lack of a genetic structure divided geographically into east-west groupings that is seen in other species from this region. [ABSTRACT FROM AUTHOR]

The ability to change reproductive tactics during adult development in response to environmental variation is predicted to enhance fitness. Many organisms show phenotypic plasticity early in nonembryonic development, but later exhibit phases of developmental inflexibility (=canalization). Therefore, we studied reproduction-related hormones and proteins and their relationships to plasticity in the Eastern lubber grasshopper. Diet-switching experiments demonstrated plasticity early in the egg production cycle, but a switch to canalization late in the cycle. We measured developmental titers of 4 hemolymph compounds from single individuals from adult molt until first oviposition. These 4 compounds were the egg-yolk precursor protein vitellogenin, juvenile hormone (the central regulator of insect reproduction), major hemolymph proteins, and ecdysteroids (the arthropod molting hormone that ultimately is stored in the egg). Using diet manipulations, we investigated how these developmental titers relate to the switch from plastic to canalized egg production. All 4 hemolymph compounds reached their peak levels during the canalized phase, about 12 day before oviposition. Diet switches after these peak levels did not affect the timing to oviposition. Therefore, these peak titers were physiological events that occurred after the individual committed to laying. We compared these patterns in reproduction to the development toward adult molt, another major life-history event in insects. We observed an extended canalized phase before the adult molt. This canalized phase always included a peak of ecdysteroids. The similar patterns in the physiology of these life-history events suggested that common limitations may exist in major developmental processes of insects that are directed by hormones. [ABSTRACT FROM AUTHOR]