Peptide pheromones in newts

This article reviews the current state of understanding of reproductive pheromones in amphibians, focusing mainly on the purification and characterization of peptide pheromones in newts of the genus Cynops, molecular cloning of cDNAs encoding the pheromone molecules, and hormonal control of secretion of these pheromones. Pheromones that attract sexually developed female Cynops pyrrhogaster and C. ensicauda newts were isolated from the male abdominal glands. The C. pyrrhogaster and C. ensicauda pheromones are peptides, designated sodefrin and silefrin, with the amino acid sequences SIPSKDALLK and SILSKDAQLK, respectively. Each pheromone attracts only conspecific females. Molecular cloning of cDNAs encoding sodefrin and silefrin revealed the presence of precursor proteins that are considered to generate these pheromone peptides. Pheromone precursor mRNA levels and radioimmunoassayable pheromone concentrations in the abdominal glands were elevated by prolactin and androgen. Sexual dimorphism and hormone dependency of the responsiveness of vomeronasal epithelium to sodefrin were noted. Significance of pheromones in the form of peptide for those performing reproductive behavior in an aquatic environment was also discussed.     

Drosophila adult and larval pheromones modulate larval food choice

Insects use chemosensory cues to feed and mate. In Drosophila, the effect of pheromones has been extensively investigated in adults, but rarely in larvae. The colonization of natural food sources by Drosophila buzzatii and Drosophila simulans species may depend on species-specific chemical cues left in the food by larvae and adults. We identified such chemicals in both species and measured their influence on larval food preference and puparation behaviour. We also tested compounds that varied between these species: (i) two larval volatile compounds: hydroxy-3-butanone-2 and phenol (predominant in D. simulans and D. buzzatii, respectively), and (ii) adult cuticular hydrocarbons (CHs). Drosophila buzzatii larvae were rapidly attracted to non-CH adult conspecific cues, whereas D. simulans larvae were strongly repulsed by CHs of the two species and also by phenol. Larval cues from both species generally reduced larval attraction and pupariation on food, which was generally—but not always—low, and rarely reflected larval response. As these larval and adult pheromones specifically influence larval food search and the choice of a pupariation site, they may greatly affect the dispersion and survival of Drosophila species in nature.     

Peptide and protein pheromones in molluscs

Pheromones have been implicated in the control of a number of behaviors in molluscs, but few peptide pheromones have been characterized in these animals. Peptide pheromones include: (1) a family of water-borne peptide pheromonal attractants (attractins) in the gastropod Aplysia that are released during egg laying and attract other Aplysia to form egg-laying and mating aggregations; (2) a tetrapeptide (ILME) in the cephalopod Sepia that elutes from egg masses and is thought to be involved in the transport of oocytes in the genital tract during egg laying; and (3) a Sepia sperm-attracting peptide (SepSAP; PIDPGVamide) that is released from oocytes during egg laying to facilitate external fertilization.     

Peptide and protein pheromones in amphibians

Purification, characterization and biological activity of urodele and anuran sex-pheromones were reviewed. Female-attracting pheromones obtained from the abdominal gland of Cynops pyrrhogaster and C. ensicauda males are peptides consisting of 10 amino acid residues being designated sodefrin and silefrin, respectively. Each pheromone attracted only conspecific females. Molecular cloning of cDNAs encoding sodefrin and silefrin revealed that both are generated from precursor proteins. Synthesis of these pheromones is regulated by prolactin (PRL) and androgen. Responsiveness of the female vomeronasal epithelium to sodefrin is enhanced by PRL and estrogen. The submandibular gland of the male terrestrial salamander, Plethodon jardani secretes a 22-kD proteinaceous pheromone that enhances female receptivity. It was revealed that every salamander synthesizes multiple isoforms of this pheromone, Plethodontid receptivity factor. The magnificent tree frog, Litoria splendida breed in an aquatic environment. The skin glands of the male secrete a female-attracting peptide pheromone, splendipherin, comprising 25 amino acid residues. The significance of the structure of the amphibian sex-pheromone as peptide and protein is discussed in terms of their species specificity.     

Peptide pheromones in female Nereis succinea

Female specimen of the ragworm, Nereis succinea, employs a tetra-peptide, cysteinyl-glutathione (CSSG) as mate recognition and gamete release pheromone during reproduction. In the present study we review the role of peptide-based pheromones in Nereid worms focusing on pheromone production in females. New results demonstrate that the female ragworms produce the pheromone in the course of the oocyte maturation directly correlated to the oocyte diameter. Nevertheless, CSSG production is significantly increasing during the reproductive process, the nuptial dance, itself. The concentrations of the pheromone, nereithione (CSSG) and those of the possible precursors, reduced glutathione (GSH) and oxidized glutathione (GSSG), in the coelomic fluid were determined by HPLC showing that during spawning the female worms excrete 68.5 +/- 12.5 microg/h of the pheromone. GSH is mobilized, or synthesized and continuously converted into the pheromone enabling female N. succinea to maintain a constant level of pheromone release until the GSH reserves are depleted. Upon release CSSG induces not only gamete release in males but in doses as low as 10(-8) to 10(-9) M also significantly increases male swimming activity and as such may guide males towards the slower swimming females.     

Peptide products of the atrial gland are not water-borne reproductive pheromones during egg laying in Aplysia

Mate attraction in Aplysia involves long-distance water-borne signaling via the secretion of the peptide pheromone attractin from the exocrine albumen gland during egg laying. Previous studies have shown that a second exocrine organ, the atrial gland, produces abundant egg-laying hormone (ELH) precursor-related peptides and mollusk-derived growth factor (MDGF), and crude extracts of the atrial gland are attractive in T-maze attraction assays. However, it is not known whether these peptides and proteins are secreted during egg laying. In this report, seawater eluates of freshly laid egg cordons were concentrated and fractionated by C18 RP-HPLC, and the resulting major peaks were examined by amino acid compositional analysis, microsequence analysis, and electrospray mass spectrometry. Concentrated egg cordon eluates were also examined by immunoblot analysis using anti-MDGF antisera as probe. The combined data demonstrated that the atrial gland of Aplysia californica does not secrete detectable levels of either ELH precursor-related peptides or MDGF during egg laying. Although the atrial gland is the last major exocrine organ to make contact with eggs before they are laid, the gland does not appear to secrete water-borne peptide pheromones during egg laying.     

Crocodile egg sounds signal hatching time

Crocodilians are known to vocalize within the egg shortly before hatching [1,2]. Although a possible function of these calls - inducing hatching in siblings and stimulating the adult female to open the nest - has already been suggested, it has never been experimentally tested [1-5]. Here, we present the first experimental evidence that pre-hatching calls of Nile crocodile (Crocodylus niloticus) juveniles are informative acoustic signals which indeed target both siblings and mother.     

Effect of flavan-3-ols on in vitro egg hatching, larval development and viability of infective larvae of Trichostrongylus colubriformis

The effects of flavan-3-ols (the monomer units of condensed tannins (CT)) and their galloyl derivatives on the viability of eggs, the development of first stage (L1) larvae, and the viability of the infective larvae of Trichostrongylus colubriformis were investigated under in vitro conditions. Each of the flavan-3-ol gallates showed some inhibition of egg hatching at 100 μg/ml, and 100% inhibition at 1000 μg/ml, with epigallocatechin gallate being the most effective in the egg hatch (EH) assay. In contrast, none of the flavan-3-ols were able to completely inhibit egg hatching. The flavan-3-ols and galloyl derivatives dose-dependently inhibited the development of infective larvae as assessed by the larval development (LD) assay. A larval migration inhibition (LMI) assay was used to assess the effect of flavan-3-ols and their galloyl derivatives on the motility of the infective third-stage (L3) larvae of T. colubriformis. In general, the flavan-3-ol gallates were more effective than the flavan-3-ols at immobilising the infective larvae as evidenced by their ability to inhibit more (P<0.05–0.01) larvae from passing through the LMI sieves. At 500 μg/ml, epigallocatechin gallate inhibited significantly more (P<0.1) larvae from passing through the sieves than did catechin gallate, epicatechin gallate, or gallocatechin gallate. Comparisons were made between the flavan-3-ols and their galloyl derivatives with the in vitro effects of CT extracts from several forage legumes, which have exhibited effects on parasites in vivo. The forage legumes tested at 200–500 μg/ml reduced the proportion of eggs that hatch, with comparable results to those obtained using the flavan-3-ols. The activities may be influenced by the prodelphinidin: procyanidin (PD:PC) ratios: CT extracts from Lotus pendunculatus and sainfoin have PD:PC ratios of 70:30 and 77:23, respectively, whereas the less active CT extract from Lotus corniculatus has a PD:PC ratio of 27:73. The active CT extracts from forage legumes have epigallocatechin as the dominant flavan-3-ol extender unit, and epigallocatechin is the most active flavan-3-ol in both the EH and LD assays.     

The relative importance of egg production rate, hatching success, hatching duration and egg sinking in population recruitment of two species of marine copepods

The disappearance of spawned copepod eggs can, at times, approach100% day^–1 and may be a bottleneck to population recruitmentof marine copepods. We examined the egg production rate andegg hatching success of Centropages hamatus and Temora longicomis(Copepoda: Calanoida) on natural diets, and the role of delayedhatching combined with high sinking rates in removing theireggs from the water column. Cumulative hatching success withinIS days was consistently high from March to June 1996:     

Synaptotagmin I functions as a calcium sensor to synchronize neurotransmitter release

To characterize Ca(2+)-mediated synaptic vesicle fusion, we analyzed Drosophila synaptotagmin I mutants deficient in specific interactions mediated by its two Ca(2+) binding C2 domains. In the absence of synaptotagmin I, synchronous release is abolished and a kinetically distinct delayed asynchronous release pathway is uncovered. Synapses containing only the C2A domain of synaptotagmin partially recover synchronous fusion, but have an abolished Ca(2+) cooperativity. Mutants that disrupt Ca(2+) sensing by the C2B domain have synchronous release with normal Ca(2+) cooperativity, but with reduced release probability. Our data suggest the Ca(2+) cooperativity of neurotransmitter release is likely mediated through synaptotagmin-SNARE interactions, while phospholipid binding and oligomerization trigger rapid fusion with increased release probability. These results indicate that synaptotagmin is the major Ca(2+) sensor for evoked release and functions to trigger synchronous fusion in response to Ca(2+), while suppressing asynchronous release.     

The complex hatching response of Aedes eggs to larval density

Abstract. 1. We investigated the effects of hatching medium, larval density and larval instar on egg hatching in Aedes triseriutus (Say) (Diptera: Cuficidae).
2. In a nutrient broth medium, hatching rates, responded positively to low densities of large larvae, but were suppressed by high densities.
3. The magnitude of suppression was moderated through the use of a yeast hatching medium, which provides a food source for the large larvae. Small instars generally exerted positive effects on hatching, with the exception of first in star larvae in the yeast medium, which inhibited hatching at low and moderate densities.
4. The results demonstrate an ability of eggs to assess indirectly the potential risks of hatching through their responses to changes in dissolved oxygen concentration.     

Induced hatching to avoid infectious egg disease in whitefish.

Reacting to a threat before physical contact, e.g., induced by air- or water-borne substances, appears to be an elegant way of defense. The reaction may be behavioral, developmental, morphological, or physiological, and it can involve a shift in niche or life history. Hatching from eggs is a shift in niche and in life history. From niche shift and life history models, one would predict that the timing of hatching is, to some degree, phenotypically plastic, i.e., early or delayed hatching is likely to be inducible. Temporary increased larval mortality (e.g., increased predation on larvae) would favor delayed hatching, while relatively high egg mortality would favor early hatching. Here, I show experimentally that eggs of the whitefish (Coregonus sp.) hatch earlier in the presence of a virulent egg parasite and that this early hatching is induced by water-borne cues emitted from infected eggs.     

Diversity of crustacean zooplankton in riparian wetlands: colonization and egg banks

Levee breaks from the Great Flood of 1993 opened up hundreds of new scour basins in the floodplain of the Missouri River. Subsequent floods, with sediment erosion and deposition, cause these lakes to be temporary features of the landscape. Within two years of the 1993 flood, the majority of zooplankton species from the region had colonized these sites. A positive correlation between species richness and connectivity indicates that sites having higher exchange with the river tended to have more species present, a result which is consistent with higher colonization rates to these sites. Hatching experiments from the sediments revealed that remnant oxbows have a highly diverse egg bank, whereas the young scour sites have limited species and numbers present. The depauperate egg bank implies that long-term population dynamics of the scours may be more dependent upon repeated colonizations than are lakes with regular emergence from the egg bank.     

广腹螳和中华大刀螳卵块孵化的比较研究

广腹螳平均卵块重和含卵量少于中华大刀螳 ,含卵量与卵块重呈正相关 ,平均孵化率分别是5 8 94%和 91 71 %。恒温条件下 ,广腹螳卵块平均孵化期分别为 5 6 2 2天 (2 0℃ ) ,3 0 71天 (2 5℃ )和 2 6 3 7天 (3 0℃ ) ,发育起始温度 1 2 65℃ ,有效积温 42 9 77日·度 ;中华大刀螳卵块平均孵化期分别为 5 0 9天(2 0℃ ) ,2 8 0 9天 (2 5℃ )和 2 1 77天 (3 0℃ ) ,发育起始温度 1 2 90℃ ,有效积温 3 62 42日·度 ;广腹螳卵块恒温孵化期短于相应变温组 ,中华大刀螳卵块恒温孵化期长于相应变温组。     

Inhibition of barnacle larval settlement and crustacean toxicity of some hoplonemertine pyridyl alkaloids

Hoplonemertines are carnivorous marine worms, which prey upon crustaceans and annelids. They paralyze their prey by injecting alkaloids with a stylet-bearing proboscis. The dermis of these animals also secretes alkaloids to repel predators. Besides affecting central and peripheral nervous system nicotinic receptors, some pyridyl alkaloids also activate certain chemoreceptor neurons in crustacean walking legs, which sense environmental chemicals. Anabaseine (2-[3-pyridyl]-3,4,5,6-tetrahydropyridyl) and 2,3'-bipyridyl (2,3'-BP) are two nemertine alkaloids, which potently paralyze crustaceans. Anabaseine is an agonist of vertebrate as well as invertebrate nicotinic receptors. While 2,3'-BP is non-toxic to mice, it is toxic to crustaceans. We tested a variety of nemertine pyridyl alkaloids for inhibition of barnacle (Balanus amphitrite) larval settlement and for crustacean toxicity in order to determine whether toxicity could be dissociated from inhibition of larval settlement. We prepared eight C-methylated 2,3'-BP isomers to determine where substitution is permitted without loss of activity. Anti-settlement and toxicity activities were not always related. For instance, 4'-methyl-2,3'-BP displayed only 3% of the crayfish paralytic activity of 2,3'-BP, but inhibited settlement almost 2-fold more effectively. Two other isomers displaying exceptional anti-settlement activity were the 4- and 5-methyl-2,3'-BPs; these also displayed high crustacean toxicity. Nemertelline inhibited barnacle settlement at concentrations similar to 2,3'-BP but was 136-fold less toxic when injected into crayfish. Thus, certain bipyridyls and tetrapyridyls may be useful anti-fouling additives.     

Omnivory in the calanoid copepod Temora longicornis: feeding, egg production and egg hatching rates

We measured in laboratory experiments the ingestion, egg production and egg hatching rates of female Temora longicornis as a function of diet. The diets consisted of a diatom (Thalassiosira weissflogii), an autotrophic dinoflagellate (Heterocapsa triquetra), and a bacterivorous ciliate (Uronema sp.) given as sole foods, or combinations of these single-food items: diatom+dinoflagellate, diatom+ciliate, dinoflagellate+ciliate, and diatom+ciliate+dinoflagellate. For the three single-item diets, the functional response was similar; i.e., ingestion rate increased linearly with food concentration (food range: ∼25 to ∼600 μg C l⁻¹). When all diets were considered, maximum daily carbon ration (∼70% of body weight) was independent of food type. However, the maximum daily egg production rate (12% of body carbon) was obtained with the diatom diet. For all diets, both ingestion and egg production rates increased with food concentration. Ingestion and egg production rates were affected differently by the interaction of food concentration and food type: at low food concentrations, ingestion rates were highest on diets containing the diatom. At high food concentrations, egg production rates were highest on the two phytoplankter diets and their combination. The presence of the ciliate in the diet did not enhance ingestion rate or egg production. Mixed-food diets did not enhance egg production relative to single-food diets. Hence, dietary diversity did not appear to be particularly advantageous for reproduction. Carbon-specific egg production efficiency (EPE; egg production/ingestion) was independent of food concentration and type, and equaled 9%. Egg hatching success was low (mean<30%) and independent of food concentration and type, and egg production rates. Our results are consistent with previous observations that egg production in T. longicornis is enhanced during diatom blooms. However, the relatively low EPE and egg hatching success suggest that reproduction and recruitment in this study were severely constrained by the biochemical composition of the diet, or the physiological condition of the females towards the end of their season of growth in Long Island Sound.