Plasticity in juvenile hormone in male burying beetles during breeding: physiological consequences of the loss of a mate

Burying beetles, Nicrophorus orbicollis, have facultative biparental care. They bury and prepare small vertebrate carcasses that provide food for their young. Here we establish the juvenile hormone (JH) profiles of paired females, paired males and single males and investigate some of the environmental and social factors that may affect these profiles. Before larvae hatch JH profiles of paired males and females were similar. However, after larvae hatch and during brood care, JH titers of females were very high and those of single males were significantly higher than those of paired males. We tested the hypothesis that higher JH was a response to the need for increased parental care by manipulating brood size. Although JH titers of single males caring for small versus large broods were not significantly different, when comparing JH titers and larval growth (a measure of parental effort), a significant positive correlation emerged. In contrast, we found that food quality had no effect on JH levels suggesting that increased feeding by males and females after carcass discovery cannot explain the elevation of JH. The regulation of JH in male burying beetles appears thus to be dependent on the presence of a mate and on critical stimuli from young.     

Social stimuli affect juvenile hormone during breeding in biparental burying beetles (Silphidae: Nicrophorus)

Extended biparental care is rare in insects but provides an excellent opportunity to investigate the interaction between the endocrine system and the physical and social environment in the regulation of this behavior. Burying beetles (Nicrophorus spp.) have facultative biparental care and depend on locating a small vertebrate carcass that they bury and prepare as food for their young. Commonly, both male and female Nicrophorus orbicollis remain in the burial chamber after eggs hatch to feed and guard the larvae. In both sexes, juvenile hormone (JH) rises rapidly in response to the discovery and assessment of the carcass; it returns to near baseline in 24 h; then in females it reaches very high titers at the onset of maternal care. In this paper, we investigate some social (presence of a mate, mating history, larval age) and environmental (carcass size) factors that may affect this endocrine profile. For females, neither the presence of a mate nor mating status (i.e., virginity) affected the initial rise of JH. However, the absence of a mate significantly depressed the JH rise in males. Eighty-seven percent of the single males buried the carcass like paired males but 87% also released pheromones to attract a mate. JH hemolymph titers in females whose broods were replaced every 24 h with newly hatch larvae were significantly higher than those of females rearing aging broods. Lastly, even though larger carcasses took longer to bury and prepare and oviposition was delayed, neither JH titers nor speed of ovarian development was affected by carcass size.     

Phe-Gly-Leu-amide allatostatin in the termite Reticulitermes flavipes: content in brain and corpus allatum and effect on juvenile hormone synthesis

In the subterranean termite Reticulitermes flavipes, allatostatins (ASTs) with the C-terminus Phe-Gly Leu-amide were localized by immunocytochemistry with antibody against a cockroach AST, Dippu AST-7. AST-immunoreactivity occurred in the corpus cardiacum and corpus allatum and in the lateral and medial neurosecretory cells of the brain that innervate these organs as well as in many other nerve cells of the brain. This was observed in workers, nymphs, soldiers and secondary reproductives. A radioimmunoassay, using anti-Dippu AST-11, demonstrated about 40 fmole equivalents of AST in brains of soldiers and secondary reproductives. The product of the corpora allata in this species was determined to be juvenile hormone III. Its synthesis by corpora allata of secondary reproductives, determined by in vitro radiochemical assay, was inhibited in a dose-dependent fashion by two cockroach allatostatins, Dippu AST-7 and Dippu AST-11. Thus, as in cockroaches and crickets, allatostatin-containing nerves innervate the corpora allata of this termite species and their production of juvenile hormone is inhibited by these neuropeptides.     

The cockroach Leucophaea maderae needs more than juvenile hormone, vitellogenin and reserves to make a yolky egg

For the cockroach Leucophaea maderae the developmental profile of lipophorin (Lp) concentrations in the hemolymph was determined through the entire vitellogenic period. At mid-vitellogenesis the concentrations of Lp had risen to 6 times the level at emergence and then declined to 2/3 of such high values at ovulation. The racemic 10R,10S-JH-III bound to Lp with an affinity of K(d) = 5.76 nM and the natural enantiomer 10R-JH-III with a K(d) = 1.60 nM. Injections of anti-Lp into mated females caused a significantly reduced rate of oocyte growth and a substantial degree of oosorption. Injections of gamma-globulin did not significantly reduce oocyte growth and caused only a small number of oocytes to resorb. Starvation after mating had similar effects as treatment with anti-Lp. Because of the high affinity of JH to Lp and since Lp occurs in micromolar concentrations during vitellogenesis one can assume that practically all JH is bound and not available for hydrolysis by the JH esterases. Lp appears to function as an inhibitor of JH metabolism by the JHEs through substrate depletion. One may conclude that a normal rate of egg growth is only achieved when titers of Lp exceed those of JH and remove major portions of this substrate from degradation by the JHEs.     

Quantification of vitellogenin-mRNA during maturation and breeding of a burying beetle

Burying beetles (Nicrophorus orbicollis) are unusual in that to breed they require an unpredictable and valuable resource, a small carcass. Thus the timing of reproduction is unpredictable and beetles' physiological response must be fast. We hypothesized that their pattern of vitellogenin (Vg) synthesis might reflect these requirements. We examined the expression of two Vg genes (sequenced for this study) during sexual maturation and through a reproductive bout. Vg-mRNA, juvenile hormone (JH) titers, ovarian development, and hemolymph concentrations of Vg were quantified in the same individuals. All four variables gradually increased during maturation to peak 15-20 days after eclosion. Twelve hours after the discovery of a carcass, a few hours before oviposition, mRNA was high, hemolymph Vg had decreased, JH and ovarian weight had increased. After oviposition, mRNA was low, hemolymph Vg concentrations and JH were high. This is consistent with our hypothesis that beetles produce and store Vg in the hemolymph prior to the discovery of a breeding resource and replace it quickly. Partial regression of these variables (with the effect of time removed) indicated that JH was not correlated with mRNA, hemolymph Vg, or ovarian weight at any time. Thus the role of JH as a gonadotropin remains unclear.     

Regulation of JH epoxide hydrolase versus JH esterase activity in the cabbage looper, Trichoplusia ni, by juvenile hormone and xenobiotics

JH III esterase and JH III epoxide hydrolase (EH) in vitro activity was compared in whole body Trichoplusia ni homogenates at each stage of development (egg, larva, pupa and adult). While activity of both enzymes was detected at all ages tested, JH esterase was significantly higher than EH activity except for day three of the fifth (last) stadium (L5D3). For both enzymes, activity was highest in eggs. Adult virgin females had 4.6- and 4.0-fold higher JH esterase and EH activities, respectively, than adult virgin males. JH III metabolic activity also was measured in whole body homogenates of fifth stadium T. ni that were fed a nutritive diet (control) or starved on a non-nutritive diet of alphacel, agar and water. With larvae that were starved for 6, 28 and 52 h, EH activity per insect equivalent was 48%, 5% and 1%, respectively, of the control insects. At the same time points, JH esterase activity levels in starved T. ni were 29%, 4% and 3% of that of insects fed the nutritive diet. Selected insect hormones and xenobiotics were administered topically or orally to fifth stadium larvae for up to 52 h, and the effects on whole body EH and JH esterase activity analyzed. JH III increased the JH III esterase activity as high as 2.2-fold, but not the JH III EH activity. The JH analog, methoprene, increased both JH esterase and EH activity as high as 2.5-fold. The JH esterase inhibitor, 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP), had no impact on EH activity. The epoxides trans- and cis-stilbene oxide (TSO and CSO) in separate experiments increased the EH activity approximately 2.0-fold. TSO did not alter JH esterase levels when topically applied, but oral administration reduced activity to 70% of the control at 28 h, and then increased the activity 1.8-fold at 52 h after the beginning of treatment. CSO had no effect on JH esterase activity. Phenobarbital increased EH activity by 1.9-fold, but did not change JH esterase levels. Clofibrate and cholesterol 5alpha,6alpha-epoxide had no effect on EH. JH esterase activity also was not affected by clofibrate, but cholesterol 5alpha,6alpha-epoxide reduced the JH esterase activity to 60-80% of the control. The biological significance of these results is discussed.     

幼年蝙蝠下丘神经元的听空间特性

实验分别在出生后４周龄的幼年和成年鲁氏菊头蝠（Ｒｈｉｎｏｌｏｐｈｕｓｒｏｕｘｉ）上进行。使用移动声刺激装置,高频喇叭可在动物头部前方水平方向１８０度、垂直方向６０度的范围内移动。玻璃微电极记录单个神经元的听反应。实验考察了幼年和成年动物下丘神经元的听空间特性,共观察了３０１个神经元,其中幼年动物１４８个,成年动物１５３个。结果表明,４周龄的幼年动物下丘听神经元已表现出方向敏感性,即每个听神经元均有一个特定的最佳反应中心和反应域。但神经元听反应中心在听空间的分布相当弥散,大多数位于对侧水平方向２０—８０度、垂直方向上下１５度范围内。而成年动物听神经元反应中心的分布则相当集中,局限地分布于对侧水平方向２８－５０度,垂直方向０—１０度范围内,两者构成明显差异。