F. Javier Sánchez-Vázquez

Design and performance of an accurate demand feeder for the study of feeding behaviour in sea bass, Dicentrarchus labrax L.

A new demand feeder was designed for the automatic and accurate feeding of fish. The feeding device is microcomputer controlled and permits the continuous recording of feeding behaviour. The electromechanical feeder consists of a sliding door powered by a solenoid which provides only one weighed pellet when activated by the fish. Some preliminary results on feeding behaviour in sea bass are presented to test the efficiency of the feeder.

Melatonin binding sites in the brain of European sea bass (Dicentrarchus labrax).

Characteristics, day-night changes, guanosine 5′-O-(3-thiotriphosphate) (GTPγS) modulation, and localization of melatonin binding sites in the brain of a marine teleost, European sea bass Dicentrarchus labrax, were studied by radioreceptor assay using 2-[125I]iodomelatonin as a radioligand. The specific binding to the sea bass brain membranes was rapid, stable, saturable and reversible. The radio-ligand binds to a single class of receptor site with the affinity (Kd) of 9.3±0.6 pM and total binding capacity (Bmax) of 39.08±0.86 fmol/mg protein (mean±SEM, n=4) at mid-light under light-dark (LD) cycles of 12:12. Day-night changes were observed neither in the Kd nor in the Bmax under LD 12:12. Treatment with GTPγS significantly increased the Kd and decreased the Bmax both at mid-light and mid-dark. The binding sites were highly specific for 2-phenylmelatonin, 2-iodomelatonin, melatonin, and 6-chloromelatonin. Distribution of melatonin binding sites in the sea bass brain was uneven: The Bmax was determined to be highest in mesencephalic optic tectum-tegmentum and hypothalamus, intermediate in telencephalon, cerebellum-vestibulolateral lobe and medulla oblongata-spinal cord, and lowest in olfactory bulbs with the Kd in the low picomolar range. These results indicate that melatonin released from the pineal organ and/or retina plays neuromodulatory roles in the sea bass brain via G protein-coupled melatonin receptors.

Ontogeny of the Circadian System During Embryogenesis in Rainbow Trout (Oncorhynchus mykyss) and the Effect of Prolonged Exposure to Continuous Illumination on Daily Rhythms of per1, clock, and aanat2 Expression

It is widely held that the development of the circadian system during embryogenesis is important for future survival of an organism. Work in teleosts has been, to date, limited to zebrafish, which provides little insight into the diversity of this system within such a large vertebrate class. In this study, the authors analyzed the diel expression of per1, clock, and aanat2 in unfertilized rainbow trout oocytes and embryos maintained under either a 12:12-h light:dark (LD) cycle or continuous illumination (LL) from fertilization. 24-h profiles in expression were measured at fertilization as well as 8, 21 42, and 57 days postfertilization (dpf). Both per1 and clock were expressed in unfertilized oocytes and all embryonic stages, whereas aanat2 expression was only measureable from 8 dpf. A reduction in both per1 and clock mean expression levels between unfertilized oocytes/0–1 dpf embryos and 8–9 dpf embryos was suggestive of a transition from maternal RNA to endogenous mRNA expression. Although aanat2 expression was not clearly associated with photic conditions, photoperiod treatment did alter the expression of per1 and clock expression/rhythmicity from as early as 8 dpf (per1), which could suggest the presence and functionality of an as yet unidentified “photoreceptor.” As a whole, this work demonstrates that clock systems are present and functional during embryonic development in rainbow trout. Further studies of their expression and regulation will help understand how the environment interacts with embryonic development in the species. (Author correspondence: andrew.davie@stir.ac.uk)

Demand feeding and welfare in farmed fish

Following the development of demand-feeding systems, many experiments have been conducted to explore feeding motivation and feed intake in farmed fish. This work aims to review a selection of studies in the field, focusing on three key factors, related to demand feeding and fish welfare. Firstly, we outline how demand feeders should be considered when developing feed management strategies for improving welfare in production conditions. Secondly, via laboratory demand-feeding experiments, we show self-feeding activities depend not only on feeding motivation and social organisation, but also on individual learning capacity and risk-taking behaviour. Thirdly, we report encouraging results demonstrating that when presented with two or more self-feeders containing complementary foods, fish select a diet according to their specific nutritional requirements, suggesting that demand feeders could be used to improve welfare by allowing fish to meet their nutritional needs.

A contact eatometer for automated continuous recording of feeding behavior in rats

A new contact eatometer designed and built in our laboratory is described. The system makes possible the reliable continuous recording of feeding behavior in rats from the time of weaning, without significantly affecting food intake- and growth-related variables. Improved time discrimination and reliability of records--along with the small financial outlay necessary to manufacture the device--make the system appropriate for chronobiological studies which involve the simultaneous use of a large number of animals.

ERK Signaling Regulates Light-Induced Gene Expression via D-Box Enhancers in a Differential, Wavelength-Dependent Manner

The day-night and seasonal cycles are dominated by regular changes in the intensity as well as spectral composition of sunlight. In aquatic environments the spectrum of sunlight is also strongly affected by the depth and quality of water. During evolution, organisms have adopted various key strategies in order to adapt to these changes, including the development of clocks and photoreceptor mechanisms. These mechanisms enable the detection and anticipation of regular changes in lighting conditions and thereby direct an appropriate physiological response. In teleosts, a growing body of evidence points to most cell types possessing complex photoreceptive systems. However, our understanding of precisely how these systems are regulated and in turn dictate changes in gene expression remains incomplete. In this manuscript we attempt to unravel this complexity by comparing the effects of two specific wavelengths of light upon signal transduction and gene expression regulatory mechanisms in zebrafish cells. We reveal a significant difference in the kinetics of light-induced gene expression upon blue and red light exposure. Importantly, both red and blue light-induced gene expression relies upon D-box enhancer promoter elements. Using pharmacological and genetic approaches we demonstrate that the ERK/MAPK pathway acts as a negative regulator of blue but not red light activated transcription. Thus, we reveal that D-box-driven gene expression is regulated via ERK/MAPK signaling in a strongly wavelength-dependent manner.

Daily rhythms of the expression of genes from the somatotropic axis: The influence on tilapia (Oreochromis niloticus) of feeding and growth hormone administration at different times.

The aim of this research was to investigate the presence of daily rhythms in the somatotropic axis of tilapia fed at two times (mid-light, ML or mid-dark, MD) and the influence of the time of day of growth hormone (GH) administration on the response of this axis. Two different GH injection times were tested: ZT 3 (3h after lights on) and ZT 15 (3h after lights off). In both experiments, the mRNA expression levels of hypothalamic pituitary adenylate cyclase-activating polypeptide (pacap), pituitary growth hormone (gh), liver insulin-like growth factors (igf1 and igf2a), and liver and muscle growth hormone receptors (ghr1 and ghr2) and IGF receptors (igf1ra and igf2r) were evaluated by means of qPCR. Daily rhythms were observed in the liver for ghr1, ghr2 and igf2r but only in fish fed at ML, with the acrophases located in the light phase (ZT 3:30, 3:31 and 7:38 h, respectively). In the muscle, ghr1 displayed a significant rhythm in both groups and ghr2 in ML fed fish (acrophases at ZT 5:29, 7:14 and 9:23h). The time of both GH administration and feeding influenced the response to GH injection: ML fed fish injected with GH at ZT 15 h showed a significant increase in liver igf1, igf2a and ghr2; and muscle ghr2 expression. This is the first report that describes the existence of daily rhythms in the somatotropic axis of tilapia and its time-dependent responses of GH administration. Our results should be considered when investigating the elements of the somatotropic axis in tilapia and GH administration.