Pheromonal control: reconciling physiological mechanism with signalling theory

Pheromones are intraspecific chemical signals. They can have profound effects on the behaviour and/or physiology of the receiver, and it is still common to hear pheromones described as controlling of the behaviour of the receiver. The discussion of pheromonal control arose initially from a close association between hormones and pheromones in the comparative physiological literature, but the concept of a controlling pheromone is at odds with contemporary signal evolution theory, which predicts that a manipulative pheromonal signal negatively affecting the receiver's fitness should not be stable over evolutionary time. Here we discuss the meaning of pheromonal control, and the ecological circumstances by which it might be supported. We argue that in discussing pheromonal control it is important to differentiate between control applied to the effects of a pheromone on a receiver's physiology (proximate control), and control applied to the effects of a pheromone on a receiver's fitness (ultimate control). Critically, a pheromone signal affecting change in the receiver's behaviour or physiology need not necessarily manipulate the fitness of a receiver. In cases where pheromonal signalling does lead to a reduction in the fitness of the receiver, the signalling system would be stable if the pheromone were an honest signal of a social environment that disadvantages the receiver, and the physiological and behavioural changes observed in the receiver were an adaptive response to the new social circumstances communicated by the pheromone.     

A review of chemical communication in freshwater fish

Since communication between individuals of a species of fish by chemical agents (pheromones) was first demonstrated in 1932, such a process has been suggested in many aspects of fish behavidïo'ur and development. This review describes observations on such mechanisms in shoaling behaviour and beneficial conditioning of water, homing of migratory fish, communication of alarm, 'crowding factor' (which adversely affects growth, survival and fecundity in dense populations), pair formation and spawning, and a range of other social interactions. Some of the chemicals involved have been isolated and identified, but most are indicated by behavioural observations. Pheromones are of great significance in fish behaviour and ecology, and are likely to be an important factor in culture operations.     

Clock-controlled endogenous melatonin rhythms in Nile tilapia (Oreochromis niloticus niloticus) and African catfish (Clarias gariepinus)

The purpose of this work was to investigate the circadian melatonin system in two tropical teleost species characterized by different behavioral habits, Nile tilapia (diurnal) and African catfish (nocturnal). To do so, fish were subjected to either a control photoperiod (12L:12D), continuous light (LL) or darkness (DD), or a 6L:6D photoperiod. Under 12L:12D, plasma melatonin levels were typically low during the photophase and high during the scotophase in both species. Interestingly, in both species, melatonin levels significantly decreased prior to the onset of light, which in catfish reached similar basal levels to those during the day, demonstrating that melatonin production can anticipate photic changes probably through circadian clocks. Further evidence for the existence of such pacemaker activity was obtained when fish were exposed to DD, as a strong circadian melatonin rhythm was maintained. Such an endogenous rhythm was sustained for at least 18 days in Nile tilapia. A similar rhythm was shown in catfish, although DD was only tested for four days. Under LL, the results confirmed the inhibitory effect of light on melatonin synthesis already reported in other species. Finally, when acclimatized to a short photo-cycle (6L:6D), no endogenous melatonin rhythm was observed in tilapia under DD, with melatonin levels remaining high. This could suggest that the circadian clocks cannot entrain to such a short photocycle. Additional research is clearly needed to further characterize the circadian axis in teleost species, identify and localize the circadian clocks, and better understand the environmental entrainment of fish physiology.     

A comparison of the activity patterns of the sand goby Pomatoschistus minutus (Pallas) from areas of different tidal range

Sand gobies ( Pomatoschistus minutus ) collected from beaches with a large tidal range (Scotland) exhibit a circatidal rhythm of activity in constant conditions in the laboratory. There is no endogenous circadian component to the rhythm. The phasing of the rhythm is such that peak activity occurs at the predicted time of ebb tide. Light-dark (LD) cycles applied in the laboratory have a marked effect on activity greatly enhancing it at night so that the original tidal rhythm becomes nocturnal. Some evidence was obtained that LD cycles can entrain a weak nocturnal circadian rhythm in fish removed from tidal conditions. Contrary to expectations, some fish from beaches with a small and unpredictable tidal range (Oslofjord, Norway) also show a weakly persistent circatidal thythm without an endogenous circadian component, but great variability was noticed between individual fish. Laboratory LD cycles did not entrain a persistent circadian rhythm in the fish from the Oslofjord.     

Biochemical properties of CikA,an unusual phytochrome-like histidine protein kinase that resets the circadian clock in Synechococcus elongatus PCC 7942

We recently described the cikA (circadian input kinase A) gene, whose product supplies environmental information to the circadian oscillator in the cyanobacterium Synechococcus elongatus PCC 7942. CikA possesses three distinct domains: a GAF, a histidine protein kinase (HPK), and a receiver domain similar to those of the response regulator family. To determine how CikA functions in providing circadian input, we constructed modified alleles to tag and truncate the protein, allowing analysis of each domain individually. CikA covalently bound bilin chromophores in vitro, even though it lacks the expected ligand residues, and the GAF domain influenced but did not entirely account for this function. Full-length CikA and truncated variants that carry the HPK domain showed autophosphorylation activity. Deletion of the GAF domain or the N-terminal region adjacent to GAF dramatically reduced autophosphorylation, whereas elimination of the receiver domain increased activity 10-fold. Assays to test phosphorelay from the HPK to the cryptic receiver domain, which lacks the conserved aspartyl residue that serves as a phosphoryl acceptor in response regulators, were negative. We propose that the cryptic receiver is a regulatory domain that interacts with an unknown protein partner to modulate the autokinase activity of CikA but does not work as bona fide receiver domain in a phosphorelay.     

INTRODUCTION: Pheromones as Tools for Olfactory Research

We have long been fascinated by the unique ability of odorsto stir our emotions and to evoke long-forgotten memories, butcertain odors play a much more fundamental role in that theyvastly improve an organism's chances for reproductive successand survival. These odorants are called pheromones, a term commonlyapplied to semiochemicals that are released by one member ofa species and evoke a specific reaction or reactions from membersof the same species. Pheromones are known for both the specificityand the potency of their actions, which can be behavioral and/orneuroendocrinological. Pheromones can stimulate individualsto aggregate, to disperse, or to react defensively in the presenceof a predator, but they are probably best known for bringingthe sexes together. Some pheromones have also been found totrigger a dramatic release of pituitary hormones in severalvertebrate species. Although first identified in insects, morerecent studies show that sex pheromones influence the livesof a wide range of organisms, from microbes to man. The hormonally-derivedsex pheromones in teleost fish, and the airborne pheromonesof moths are two systems that illustrate how scientists haveused these specialized chemical signals as important tools toinvestigate the morphology, physiology and biochemistry of olfactory-receptorsystems, the mechanisms of odor-information processing in thebrain, and the diverse range of behaviors and endocrinologicalchanges associated with pheromonal communication. While ourfocus is on these two animal models, other examples, includingmammalian pheromone systems, are also discussed. Chem. Senses21: 241–243, 1996.     

Information Transfer across Fish Shoals under Predator Threat

The hypothesis that shoaling fish can obtain information about a predator's approach from changes in the behaviour of other shoal members was tested in an experiment in which receiver minnows (Phoxinus phoxinus) behind a one-way-mirror could observe the reaction of transmitter minnows threatened by the stalk of a pike (Esox lucius) model. Although the receiver minnows were out of visual contact with the pike model they decreased their foraging behaviour and started hiding when the pike model came towards the feeding patch of the transmitter fish. The presence of skittering behaviour and inspection behaviour in the transmitter but not in the receiver fish suggests that individual confirmation of the predator does play a role in determining the nature of the anti-predator response. Nevertheless there is clear evidence that receiver fish modify their own behaviour on the basis of information obtained from individuals which have seen a predator.     

The Neurospora crassa pheromone precursor genes are regulated by the mating type locus and the circadian clock

Pheromones play important roles in female and male behaviour in the filamentous ascomycete fungi. To begin to explore the role of pheromones in mating, we have identified the genes encoding the sex pheromones of the heterothallic species Neurospora crassa. One gene, expressed exclusively in mat A strains, encodes a polypeptide containing multiple repeats of a putative pheromone sequence bordered by Kex2 processing sites. Strains of the opposite mating type, mat a, express a pheromone precursor gene whose polypeptide contains a C-terminal CAAX motif predicted to produce a mature pheromone with a C-terminal carboxy-methyl isoprenylated cysteine. The predicted sequences of the pheromones are remarkably similar to those encoded by other filamentous ascomycetes. The expression of the pheromone precursor genes is mating type specific and is under the control of the mating type locus. Furthermore, the genes are highly expressed in conidia and under conditions that favour sexual development. Both pheromone precursor genes are also regulated by the endogenous circadian clock in a time-of-day-specific fashion, supporting a role for the clock in mating.     

Influence of light and food on the circadian clock in liver of rainbow trout,Oncorhynchus mykiss

Several reports support the existence of multiple peripheral oscillators in fish, which may be able to modulate the rhythmic functions developed by those tissues hosting them. Thus, a circadian oscillator has been proposed to be located within fish liver. In this vertebrate group, the role played by the circadian system in regulating metabolic processes in liver is mostly unknown. We, therefore investigated the liver of rainbow trout (Oncorhynchus mykiss) as a potential element participating in the regulation of circadian rhythms in fish by hosting a functional circadian oscillator. The presence and expression pattern of main components of the circadian molecular machinery (clock1a, bmal1, per1 and rev-erbβ-like) were assessed. Furthermore, the role of environmental cues such as light and food, and their interaction in order to modulate the circadian oscillator was also assessed by exposing animals to constant conditions (absence of light for 48 h, and/or a 4 days fasting period). Our results demonstrate the existence of a functional circadian oscillator within trout liver, as demonstrated by significant rhythms of all clock genes assessed, independently of the environmental conditions studied. In addition, the daily profile of mRNA abundance of clock genes is influenced by both light (mainly clock1a and per1) and food (rev-erbβ-like), which is indicative of an interaction between both synchronizers. Our results point to rev-erbβ-like as possible mediator between the influence of light and food on the circadian oscillator within trout liver, since its daily profile is influenced by both light and food, thus affecting that of bmal1.     

Circadian Rhythms of Locomotor Activity in the Nile Tilapia Oreochromis niloticus

Behavioral rhythms of the Nile tilapia were investigated to better characterize its circadian system. To do so, the locomotor activity patterns of both male and female tilapia reared under a 12:12 h light-dark (LD) cycle were studied, as well as in males the existence of endogenous rhythmicity under free-running conditions (DD and 45 min LD pulses). When exposed to an LD cycle, the daily pattern of activity differed between individuals: some fish were diurnal, some nocturnal, and a few displayed an arrhythmic pattern. This variability would be typical of the plastic circadian system of fish. Moreover, reproductive events clearly affected the behavioral rhythms of female tilapia, a mouth-brooder teleost species. Under DD, 50% (6 of 12) of male fish showed circadian rhythms with an average period (τ) of 24.1±0.2 h, whereas under the 45 min LD pulses, 58% (7 of 12) of the fish exhibited free-running activity rhythms with an average τ of 23.9±0.5 h. However, interestingly in this case, activity was always confined to the dark phase. Furthermore, when the LD cycle was reversed, a third of the fish showed gradual resynchronization to the new phase, taking 7–10 days to be completely re-entrained. Taken together, these results suggest the existence of an endogenous circadian oscillator that controls the expression of locomotor activity rhythms in the Nile tilapia, although its anatomical localization remains unknown.     

Photic and circadian regulation of melatonin production in the Mozambique tilapia Oreochromis mossambicus

Diverse circadian systems related to phylogeny and ecological adaptive strategies are proposed in teleosts. Recently, retinal photoreception was reported to be important for the circadian pacemaking activities of the Nile tilapia Oreochromis niloticus. We aimed to confirm the photic and circadian responsiveness of its close relative-the Mozambique tilapia O. mossambicus. Melatonin production in cannulated or ophthalmectomized fish and its secretion from cultured pineal glands were examined under several light regimes. Melatonin production in the cannulated tilapias was measured at 3-h intervals; it fluctuated daily, with a nocturnal increase and a diurnal decrease. Exposing the cannulated fish to several light intensities (1500-0.1 lx) and to natural light (0.1 and 0.3 lx) suppressed melatonin levels within 30 min. Static pineal gland culture under light-dark and reverse light-dark cycles revealed that melatonin synthesis increased during the dark periods. Rhythmic melatonin synthesis disappeared on pineal gland culture under constant dark and light conditions. After ophthalmectomy, plasma melatonin levels did not vary with light-dark cycles. These results suggest that (1) Mozambique tilapias possess strong photic responsiveness, (2) their pineal glands are sensitive to light but lack circadian pacemaker activity, and (3) they require lateral eyes for rhythmic melatonin secretion from the pineal gland.     

Light and dark rations and the photic entrainment of circadian locomotor activity patterns in the South American Silver Catfish (Rhamdia quelen,Quoy & Gaimard, 1824)

The aim of this study was to evaluate the daily rhythm of locomotor activity in Rhamdia quelen (R. quelen). A total of 30 fish were enrolled in the study and were equally divided in 10 groups and maintained in 100 liters tanks. The locomotor activity was measured in fish maintained under the LD 12:12 photoperiod regime; thereafter, the LD cycle was reversed to DL in order to study the resynchronization and to explore the endogenous pacemaker. Subsequently, the fish were subjected to constant conditions of light to test whether or not locomotor rhythms are regulated by the endogenous circadian clock. The effect of increasing light length and intensity was studied on daily rhythm of locomotor activity of fish. Our results showed that the R. quelen is a strictly diurnal species, the rhythm of locomotory activity resynchronized quickly after inverting the LD cycle and persist under free course LL, suggesting a circadian origin. The light showed a significant masking effect often blocking the expression of the biological rhythm. The strictly diurnal behavior is controlled directly by the photoperiod and maintained even under very dim light (30 lux).     

Early development of circadian rhythmicity in the suprachiamatic nuclei and pineal gland of teleost,flounder (Paralichthys olivaeus), embryos

Circadian rhythms enable organisms to coordinate multiple physiological processes and behaviors with the earth's rotation. In mammals, the suprachiasmatic nuclei (SCN), the sole master circadian pacemaker, has entrainment mechanisms that set the circadian rhythm to a 24‐h cycle with photic signals from retina. In contrast, the zebrafish SCN is not a circadian pacemaker, instead the pineal gland (PG) houses the major circadian oscillator. The SCN of flounder larvae, unlike that of zebrafish, however, expresses per2 with a rhythmicity of daytime/ON and nighttime/OFF. Here, we examined whether the rhythm of per2 expression in the flounder SCN represents the molecular clock. We also examined early development of the circadian rhythmicity in the SCN and PG. Our three major findings were as follows. First, rhythmic per2 expression in the SCN was maintained under 24 h dark (DD) conditions, indicating that a molecular clock exists in the flounder SCN. Second, onset of circadian rhythmicity in the SCN preceded that in the PG. Third, both 24 h light (LL) and DD conditions deeply affected the development of circadian rhythmicity in the SCN and PG. This is the first report dealing with the early development of circadian rhythmicity in the SCN in fish.     

Circadian rhythms and photic entrainment of swimming activity in cave-dwelling fish Astyanax mexicanus (Actinopterygii: Characidae), from El Sotano La Tinaja,San Luis Potosi,Mexico

Circadian regulation has a profound adaptive meaning in timing the best performance of biological functions in a cyclic niche. However, in cave-dwelling animals (troglobitic), a lack of photic cyclic environment may represent a disadvantage for persistence of circadian rhythms. There are different populations of cave-dwelling fish Astyanax mexicanus in caves of the Sierra El Abra, Mexico, with different evolutive history. In the present work, we report that fish collected from El Sótano la Tinaja show circadian rhythms of swimming activity in laboratory conditions. Rhythms observed in some of the organisms entrain to either continuous light–dark cycles or discrete skeleton photoperiods tested. Our results indicate that circadian rhythm of swimming activity and their ability to entrain in discrete and continuous photoperiods persist in some organisms that might represent one of the oldest populations of cave-dwelling A. mexicanus in the Sierra El Abra.     

Eyeless Mexican Cavefish Save Energy by Eliminating the Circadian Rhythm in Metabolism

The eyed surface form and eyeless cave form of the Mexican tetra Astyanax mexicanus experience stark differences in the daily periodicities of light, food and predation, factors which are likely to have a profound influence on metabolism. We measured the metabolic rate of Pachón cave and surface fish at a fixed swimming speed under light/dark and constant dark photoperiods. In constant darkness surface forms exhibited a circadian rhythm in metabolism with an increase in oxygen demand during the subjective daytime, whereas cave forms did not. The lack of circadian rhythm in metabolism leads to a 27% energy savings for Pachón cave fish compared to surface fish when comparing both forms in their natural photoperiods. When surface forms were tested under constant dark conditions they expended 38% more energy than cave forms under equivalent conditions. Elimination of the circadian rhythm in metabolism may be a general feature of animals that live in perpetually dark food-limited environments such as caves or the deep sea.     

Pheromones and signature mixtures: defining species-wide signals and variable cues for identity in both invertebrates and vertebrates

Pheromones have been found in species in almost every part of the animal kingdom, including mammals. Pheromones (a molecule or defined combination of molecules) are species-wide signals which elicit innate responses (though responses can be conditional on development as well as context, experience, and internal state). In contrast, signature mixtures, in invertebrates and vertebrates, are variable subsets of molecules of an animal’s chemical profile which are learnt by other animals, allowing them to distinguish individuals or colonies. All signature mixtures, and almost all pheromones, whatever the size of molecules, are detected by olfaction (as defined by receptor families and glomerular processing), in mammals by the main olfactory system or vomeronasal system or both. There is convergence on a glomerular organization of olfaction. The processing of all signature mixtures, and most pheromones, is combinatorial across a number of glomeruli, even for some sex pheromones which appear to have ‘labeled lines’. Narrowly specific pheromone receptors are found, but are not a prerequisite for a molecule to be a pheromone. A small minority of pheromones act directly on target tissues (allohormone pheromones) or are detected by non-glomerular chemoreceptors, such as taste. The proposed definitions for pheromone and signature mixture are based on the heuristic value of separating these kinds of chemical information. In contrast to a species-wide pheromone, there is no single signature mixture to find, as signature mixtures are a ‘receiver-side’ phenomenon and it is the differences in signature mixtures which allow animals to distinguish each other.     

Fatty acid levels in blood of cyclostomes and fish

Synopsis Experimental data obtained on cyclostomes and fish concerning the levels of plasma fatty acids (PFA), their biological role as an energetic substrate, various factors affecting PFA and mechanisms involved in the control of their level are reviewed.It was shown that PFA levels in lamprey and fish depend strongly on season, circadian rhythm, nutritional state, age, sex, intrinsic activity of individuals and are regulated by some metabolites as well as hormones. The question concerning the nervous control of PFA in cyclostomes and fish needs further elucidation on account of scarce information available at present.     

Photic entrainment of circadian activity patterns in the tropical labrid fish Halichoeres chrysus

Yellow wrasses (Halichoeres chrysus) show clear daily activity patterns. The fish hide in the substrate at (subjective) night, during the distinct rest phase. Initial entrainment in a 12h:12h light-dark (12:12 LD) cycle (mean period 24.02h, SD 0.27h, n = 16 was followed by a free run (mean period 24.42h, SD 1.33h) after transition into constant dim light conditions. Light pulses of a comparable intensity as used in the light part of the LD cycles did not result in significant phase shifts of the free-running rhythm in constant darkness. Application of much brighter 3h light pulses resulted in a phase-response curve (PRC) for a fish species, with pronounced phase advances during late subjective night. The PRCs differed from those mainly obtained in other vertebrate taxa by the absence of significant phase delays in the early subjective night. At that circadian phase, significant tonic effects of the light pulses caused a shortening of the circadian period length. Entrainment to skeleton photoperiods of 1:11 LD was observed in five of six wrasses exposed, also after a 3h phase advance of this LD cycle. Subsequently, a 1:11.25 LD cycle resulted in entrainment in four of the six fish. It is suggested that the expression of the circadian system in fish can be interpreted as a functional response to a weak natural zeitgeber, as present in the marine environment. This response allows photic entrainment as described here in the yellow wrasse. (Chronobiology International, 17(5), 613-622, 2000)     

Sexual attraction: on the role of fungal pheromone/receptor systems (A review)

Pheromones have been detected in all fungal phylogenetic lineages. This came as a surprise, as the general role of pheromones in mate attraction was not envisioned for some fungi. Pheromones and pheromone receptor genes have been identified, however, in members of all true fungal lineages, and even for mycelia forming organisms of plant and amoeba lineages, like oomycetes and myxomycetes. The mating systems and genes governing the mating type are different in fungi, ranging from bipolar with two opposite mating types to tetrapolar mating systems (with four possible mating outcomes, only one of which leads to fertile sexual development) in homobasidioymcetes with more than 23,000 mating types occurring in nature. Pheromones and receptors specifically recognizing these pheromones have evolved with slightly different functions in these different systems. This review is dedicated to follow the evolution of pheromone/receptor systems from simple, biallelic bipolar systems to multiallelic, tetrapolar versions and to explain the slightly different functions the pheromone recognition and subsequent signal transduction cascades within the fungal kingdom. The biotechnological implications of a detailed understanding of mating systems for biological control and plant protection, in medicine, and in mushroom breeding are discussed.     

A biotelemetry system recording fish activity

A biotelemetry system is described for obtaining, transmitting and recording the electromyograms (EMGs) produced in muscle activity of free-swimming fish as quantitative indicators of overall fish activity. The radiotransmitters used come in the form of cylindrical packages having two sensing electrodes, all fully implantable in the fish body cavity. EMGs are transmitted as radio pulses with the intensity of muscular activity determining the intervals between pulses. The packages also contain temperature sensors and fish temperatures are transmitted with every 32nd pulse. Transmitted EMG pulses are detected, 'measured' and stored by a single portable receiver (Model SRX_400, Lotek). Data can be subsequently transferred to a computer (which can also be portable) for storage, processing and statistical analysis. Transmitter battery life can be in excess of 7 months, permitting laboratory or field studies of long duration. Transmitter package implantation surgery requires a mid-ventral incision and internal securing of transmitter and sensing electrodes. Surgical silk, cyanoacrylate tissue adhesives, and polydioxanone (PD), a synthetic absorbable suture, were all tried as means of incision closure. The most effective was PD alone. Trials of the system consisted of forced swims by transmitter-equipped rainbow trout Oncorhynchus mykiss Walbaum. The data obtained provided an inverse linear relation between forced swim speed and EMG pulse interval. Trials were conducted at intervals over periods up to 2 months. Fish showed neither distress, nor difficulty in swimming up to maximum speeds of 60 cm s ⁻¹ (fish lengths 41.0, 44.4 cm).