Vincent Gennotte

Characterization of the primary sonic muscles in Carapus acus (Carapidae): a multidisciplinary approach

Sound production in carapid fishes results from the action of extrinsic muscles that insert into the swim bladder. Biochemical, histochemical and morphological techniques were used to examine the sonic muscles and compare them with epaxial muscles in Carapus acus. Sonic fibres are thicker than red and thinner than white epaxial fibres, and sonic fibres and myofibrils exhibit an unusual helicoidal organization: the myofibrils of the centre are in a straight line whereas they are more and more twisted towards the periphery. Sonic muscles have both features of red (numerous mitochondria, high glycogen content) and white (alkali–stable ATPase) fibres. They differ also in the isoforms of the light chain (LC3) and heavy chain (HC), in having T tubules at both the Z–line and the A–I junction and in a unique parvalbumin isoform (PAI) that may aid relaxation. All these features lead to the expression of two assumptions about sound generation: the sonic muscle should be able to perform fast and powerful contractions that provoke the forward movement of the forepart of the swim bladder and the stretching and ‘flapping’ of the swim bladder fenestra; the helicoidal organization allows progressive drawing of the swim bladder fenestra which emits a sound when rapidly released in a spring–like manner.

Behaviours Associated with Acoustic Communication in Nile Tilapia (Oreochromis niloticus)

Background Sound production is widespread among fishes and accompanies many social interactions. The literature reports twenty-nine cichlid species known to produce sounds during aggressive and courtship displays, but the precise range in behavioural contexts is unclear. This study aims to describe the various Oreochromis niloticus behaviours that are associated with sound production in order to delimit the role of sound during different activities, including agonistic behaviours, pit activities, and reproduction and parental care by males and females of the species. Methodology/Principal Findings Sounds mostly occur during the day. The sounds recorded during this study accompany previously known behaviours, and no particular behaviour is systematically associated with sound production. Males and females make sounds during territorial defence but not during courtship and mating. Sounds support visual behaviours but are not used alone. During agonistic interactions, a calling Oreochromis niloticus does not bite after producing sounds, and more sounds are produced in defence of territory than for dominating individuals. Females produce sounds to defend eggs but not larvae. Conclusion/Significance Sounds are produced to reinforce visual behaviours. Moreover, comparisons with O. mossambicus indicate two sister species can differ in their use of sound, their acoustic characteristics, and the function of sound production. These findings support the role of sounds in differentiating species and promoting speciation. They also make clear that the association of sounds with specific life-cycle roles cannot be generalized to the entire taxa.

The sensitive period for male-to-female sex reversal begins at the embryonic stage in the Nile tilapia and is associated with the sexual genotype

In this study, we sought to determine the mechanism of early sex reversal in a teleost by applying 4 hr feminization treatments to XY (17α‐ethynylestradiol 2000 μg L−1) and YY (6500 μg L−1) Nile tilapia embryos on the first day post‐fertilization (dpf). We then searched for changes in the expression profiles of some sex‐differentiating genes in the brain (cyp19a1b, foxl2, and amh) and in sex steroids (testosterone, 17β‐estradiol, and 11‐ketotestosterone) concentrations during embryogenesis and gonad differentiation. No sex reversal was observed in YY individuals, whereas sex‐reversal rates in XY progeny ranged from 0–60%. These results, together with the clearance profile of 17α‐ethynylestradiol, confirmed the existence of an early sensitive period for sex determination that encompasses embryonic and larval development and is active prior to any sign of gonad differentiation. Estrogen treatment induced elevated expression of cyp19a1b and higher testosterone and 17β‐estradiol concentrations at 4 dpf in both XY and YY individuals. foxl2 and amh were repressed at 4 dpf and their expression levels were not different between treated and control groups at 14 dpf, suggesting that foxl2 did not control cyp19a1b in the brains of tilapia embryos. Increased cyp19a1b expression in treated embryos could reflect early brain sexualization, although this difference alone cannot account for the observed sex reversal as the treatment was ineffective in YY individuals. The differential sensitivity of XY and YY genotypes to embryonic induced‐feminization suggests that a sex determinant on the sex chromosomes, such as a Y repressor or an X activator, may influence sex reversal during the first steps of tilapia embryogenesis. Mol. Reprod. Dev. 81: 1146–1158, 2014.

Brief exposure of embryos to steroids or aromatase inhibitor induces sex reversal in Nile tilapia (Oreochromis niloticus).

This study aimed to develop sex reversal procedures targeting the embryonic period as tools to study the early steps of sex differentiation in Nile tilapia with XX, XY, and YY sexual genotypes. XX eggs were exposed to masculinizing treatments with androgens (17α-methyltestosterone, 11-ketotestosterone) or aromatase inhibitor (Fadrozole), whereas XY and YY eggs were subjected to feminizing treatments with estrogen analog (17α-ethynylestradiol). All treatments consisted of a single or double 4-hr immersion applied between 1 and 36 hour post-fertilization (hpf). Concentrations of active substances were 1000 or 2000 μg l(-1) in XX and XY, and 2000 or 6500 μg l(-1) in YY. Masculinizing treatments of XX embryos achieved a maximal sex reversal rate of 10% with an exposure at 24 hpf to 1000 μg l(-1) of 11-ketotestosterone or to 2000 μg l(-1) of Fadrozole. Feminization of XY embryos was more efficient and induced up to 91% sex reversal with an exposure to 2000 μg l(-1) of 17α-ethynylestradiol. Interestingly, similar treatments failed to reverse YY fish to females, suggesting either that a sex determinant linked to the Y chromosome prevents the female pathway when present in two copies, or that a gene present on the X chromosome is needed for the development of a female phenotype.

Ontogeny of Swimming Movements in the Catfish Clarias gariepinus

The swimming movements of C. gariepinus larvae were recorded with a high-speed camera (400, 500 and 800 fps) from 0 to 336 hours post-hatching. Movements of adult fish were also recorded to provide information on the last developmental stage. Seven landmarks positioned on the fish midline were used during tail beating to determine various parameters during ontogeny and, on the basis of these parameters, to describe the first appearance of swimming move- ments and their development and efficiency during growth. Larvae were unable to swim at hatching (4 mm total length). Swimming movements were established at 48 hours post- hatching when the fish measured between 7 and 8 mm total length and the yolk sac was more than 95% absorbed. At this stage, lateral excursion of the head appeared strongly reduced (from 13% to 6% of the total length). The efficiency of swimming movements increased throughout ontogeny, as did the homogeneity of the speed of the propulsive wave. Spon- taneous swimming speed of 1 to 10 TLs -1 were observed in early stage (8-12 hPH). The various speed induced significant variations in parameters such as the amplitude of lateral head movements, swimming efficiency, and body rigidity. No major change was observed at the theoretical flow-regime transition.

Temperature Preference and Sex Differentiation in African Catfish, Clarias gariepinus

The African catfish Clarias gariepinus has a genetic sex determination system in which high temperature induces masculinization. The thermosensitive period for sex differentiation is short and occurs very early (from 6 to 8 days posthatching [dph]). As young juveniles can encounter high masculinizing temperature (36.5°C) in African water points, we aimed to determine the thermal preference of sexually undifferentiated juveniles and investigate if they spontaneously move toward high masculinizing temperature. Experiments were carried out in an environmental continuum (28-28-28°C and 28-32-36.5°C) made up of three 50-L aquariums connected together. Four hundred larvae from 10 different full-sib progenies were reared successively from 2 to 14 dph in these facilities. Before and after thermal treatments, fish were reared at 28°C until sex ratio determination at 70 dph. In the control continuum, fish were nearly equally distributed in the three compartments. Conversely, in the thermal continuum, compartment occupation significantly differed with progeny and period. During the highly thermosensitive period, two of five progenies significantly preferred (54.7% and 39.8% occupation) the 36.5°C compartment. All tested progenies reared in thermal continuum and separated 36.5°C aquarium showed a skewed sex ratio toward the male phenotype (78-100%). Nevertheless, no correlation was found between 36.5°C compartment occupation and sex ratio in thermal continuum groups. As masculinization temperature could be encountered in African water points during the spawning season, we discussed the adaptive advantage for the African catfish to display a sex differentiation process controlled by a temperature effect.

Do sex reversal procedures differentially affect agonistic behaviors and sex steroid levels depending on the sexual genotype in Nile tilapia

In Nile tilapia Oreochromis niloticus, phenotypic males and females with different sexual genotypes (XX, XY, YY) have particular behavioral and physiological traits. Compared to natural XX females and XY males, XY and YY females and XX males expressed higher level of aggressiveness that could be related to higher levels of 17β-estradiol and 11-ketotestosterone, respectively. Our results suggest that the presence of a Y chromosome increases aggressiveness in females. However, since the same relationship between aggressiveness and the Y chromosome is not observed in males, we can hypothesize that the differences in aggressiveness are not directly dependent on the genotype but on the sex reversal procedures applied on young fry during their sexual differentiation to produce these breeders. These hormonal treatments could have permanently modified the development of the brain and consequently influenced the behavior of adults independently of their genotype. In both hypotheses (genotype or sex reversal influence), the causes of behavioral modifications have to be searched in an early modification of the brain sexual differentiation.