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Smoltification prepares juvenile Atlantic salmon (Salmo salar) for downstream migration. Dramatic changes characterize this crucial event in the salmon’s life cycle, including increased gill Na+/K+-ATPase activity (NKA) and plasma hormone levels. The triggering of smoltification relies on photoperiod and is modulated by temperature. Both provide reliable information, to which fish have adapted for thousands of years, that allows deciphering daily and calendar time. Here we studied the impact of different photoperiod (natural, sustained winter solstice) and temperature (natural, ~ + 4° C) combinations, on gill NKA, plasma free triiodothyronine (T3) and thyroxine (T4), and melatonin (MEL; the time-keeping hormone), throughout smoltification. We also studied the impact of temperature history on pineal gland MEL production in vitro. The spring increase in gill NKA was less pronounced in smolts kept under sustained winter photoperiod and/or elevated temperature. Plasma thyroid hormone levels displayed day–night variations, which were affected by elevated temperature, either independently from photoperiod (decrease in T3 levels) or under natural photoperiod exclusively (increase in T4 nocturnal levels). Nocturnal MEL secretion was potentiated by the elevated temperature, which also altered the MEL profile under sustained winter photoperiod. Temperature also affected pineal MEL production in vitro, a response that depended on previous environmental acclimation of the organ. The results support the view that the salmon pineal is a photoperiod and temperature sensor, highlight the complexity of the interaction of these environmental factors on the endocrine system of S. salar, and indicate that climate change might compromise salmon’s time “deciphering” during smoltification, downstream migration and seawater residence.

Background and Purpose— Mexican Americans (MAs) have worse neurological, functional, and cognitive outcomes after stroke. Stroke rehabilitation is important for good outcome. In a population-based study, we sought to determine whether allocation of stroke rehabilitation services differed by ethnicity. Methods— Patients with stroke were identified as part of the Brain Attack Surveillance in Corpus Christi (BASIC) project, TX, USA. Cases were validated by physicians using source documentation. Patients were followed prospectively for 3 months after stroke to determine rehabilitation services and transitions. Descriptive statistics were used to depict the study population. Continuous baseline variables were compared using 2 sample t tests or Wilcoxon rank-sum tests by ethnicity. Categorical baseline variables were compared using χ 2 tests. Ethnic comparisons of rehabilitation services were compared using χ 2 tests, Fisher’s exact tests, and logistic regression. Results— Seventy-two subjects (50 MA and 22 non-Hispanic white [NHW]) were followed. Mean age, NHW-69 (SD 13), MA-66 (SD 11) years, sex (NHW 55% male, MA 50% male) and median presenting National Institutes of Health Stroke Scale did not differ significantly. There were no ethnic differences among the proportion of patients who were sent home without any rehabilitation services ( P =0.9). Among those who received rehabilitation, NHWs were more likely to get inpatient rehabilitation (73%) compared with MAs (30%), P =0.016. MAs (51%) were much more likely to receive home rehabilitation services compared with NHWs (0%) ( P =0.0017). Conclusions— In this population-based study, MAs were more likely to receive home-based rehabilitation, whereas NHWs were more likely to get inpatient rehabilitation. This disparity may, in part, explain the worse stroke outcome in MAs.

International audience; Melatonin is an important component of the vertebrates circadian system, synthetized from serotonin by the successive action of the arylalkylamine N-acetyltransferase (Aanat: serotonin[RIGHTWARDS ARROW]N-acetylserotonin) and acetylserotonin-O-methyltransferase (Asmt: N-acetylserotonin[RIGHTWARDS ARROW]melatonin). Aanat is responsible for the daily rhythm in melatonin production. Teleost fish are unique because they express two Aanat genes, aanat1 and aanat2, mainly expressed in the retina and pineal gland, respectively. In silico analysis indicated that the teleost-specific whole-genome duplication generated Aanat1 duplicates (aanat1a and aanat1b); some fish express both of them, while others express either one of the isoforms. Here, we bring the first information on the structure, function, and distribution of Aanat1a and Aanat1b in a teleost, the sea bass Dicentrarchus labrax. Aanat1a and Aanat1b displayed a wide and distinct distribution in the nervous system and peripheral tissues, while Aanat2 appeared as a pineal enzyme. Co-expression of Aanats with asmt was found in the pineal gland and the three retinal nuclear layers. Enzyme kinetics indicated subtle differences in the affinity and catalytic efficiency of Aanat1a and Aanat1b for indolethylamines and phenylethylamines, respectively. Our data are consistent with the idea that Aanat2 is a pineal enzyme involved in melatonin production, while Aanat1 enzymes have a broader range of functions including melatonin synthesis in the retina, and catabolism of serotonin and dopamine in the retina and other tissues. The data are discussed in light of the recently uncovered roles of N-acetylserotonin and N-acetyldopamine as antioxidants, neuroprotectants, and modulators of cell proliferation and enzyme activities.

Background: Mexican Americans (MAs) have worse neurologic, functional and cognitive outcomes after stroke than non Hispanic whites (NHWs). Stroke rehabilitation is important for outcome. In a population-based study, we sought to determine if allocation of stroke rehabilitation services differed by ethnicity. Methods: Consecutive stroke patients were identified for a three month time period as part of the Brain Attack Surveillance in Corpus Christi (BASIC) project, Texas, USA. Cases were validated by physicians using source documentation. Patients were followed prospectively for three months following stroke to determine self-reported rehabilitation services. Descriptive statistics were used to describe the study population. Ethnic comparisons of rehabilitation services were made using chi-squared or Fisher’s exact tests. Results: Seventy-two subjects (50 MA, 22 NHW) were followed. Mean age, NHW-69 (sd-13), MA-66 (sd-11) years, sex (NHW 55% male, MA 50% male) and median presenting NIHSS (NHW-2.5, MA-3.0) did not differ significantly. There were no ethnic differences in the proportion of patients who were discharged home without rehabilitation services (p=0.9). Among those who received rehabilitation (n=48), the figure shows the distribution of the first place for services. NHWs were more likely to be discharged to inpatient rehabilitation (73%) compared with MAs (30%), p=0.016. MAs (51%) were much more likely to be receive home rehabilitation services compared with NHWs (0%) (p=0.0017). Conclusions: In this population-based study, MAs were more likely to receive home-based rehabilitation while NHWs more likely to get inpatient rehabilitation. This disparity may, in part, explain the worse stroke outcome in MAs.

The master circadian clock in fish has been considered to reside in the pineal gland. This dogma is challenged, however, by the finding that most zebrafish tissues contain molecular clocks that are directly reset by light. To further examine the role of the pineal gland oscillator in the zebrafish circadian system, we generated a transgenic line in which the molecular clock is selectively blocked in the melatonin-producing cells of the pineal gland by a dominant-negative strategy. As a result, clock-controlled rhythms of melatonin production in the adult pineal gland were disrupted. Moreover, transcriptome analysis revealed that the circadian expression pattern of the majority of clock-controlled genes in the adult pineal gland is abolished. Importantly, circadian rhythms of behavior in zebrafish larvae were affected: rhythms of place preference under constant darkness were eliminated, and rhythms of locomotor activity under constant dark and constant dim light conditions were markedly attenuated. On the other hand, global peripheral molecular oscillators, as measured in whole larvae, were unaffected in this model. In conclusion, characterization of this novel transgenic model provides evidence that the molecular clock in the melatonin-producing cells of the pineal gland plays a key role, possibly as part of a multiple pacemaker system, in modulating circadian rhythms of behavior., Author Summary Most physiological and behavioral processes exhibit daily rhythms which are driven by an internal timing mechanism known as the circadian clock. Circadian systems are thought to be organized in a hierarchical manner, with a central pacemaker in the brain regulating peripheral clocks throughout the body. In fish species, the pineal gland has been considered to function as a central circadian clock organ. Nevertheless, a central role for the pineal gland in the zebrafish circadian system has been questioned, because peripheral clocks in this species are independently synchronized by light. Here we developed a genetically modified zebrafish model in which the molecular clock is selectively blocked in the melatonin-producing cells of the pineal gland. As a result, clock-controlled melatonin production and gene expression in the pineal gland are interrupted. Although the independent peripheral clocks are not affected by this genetic manipulation, circadian rhythms of behavior are attenuated. These findings indicate that the zebrafish pineal gland clock contributes to the generation of daily behavioral rhythms, possibly as part of a multiple pacemaker system.

In most species daily rhythms are synchronized by the photoperiodic cycle. They are generated by the circadian system, which is made of a pacemaker, an entrainment pathway to this clock, and one or more output signals. In vertebrates, melatonin produced by the pineal organ is one of these outputs. The production of this time-keeping hormone is high at night and low during the day. Despite the fact that this is a well-preserved pattern, the pathways through which the photoperiodic information controls the rhythm have been profoundly modified from early vertebrates to mammals. The photoperiodic control is direct in fish and frogs and indirect in mammals. In the former, full circadian systems are found in photoreceptor cells of the pineal organ, retina, and possibly brain, thus forming a network where melatonin could be a hormonal synchronizer. In the latter, the three elements of a circadian system are scattered: the photoreceptive units are in the eyes, the clocks are in the suprachiasmatic nuclei of the hypothalamus, and the melatonin-producing units are in the pineal cells. Intermediate situations are observed in sauropsids. Differences are also seen at the level of the arylalkylamine N-acetyltransferase (AANAT), the enzyme responsible for the daily variations in melatonin production. In contrast to tetrapods, teleost fish AANATs are duplicated and display tissue-specific expression; also, pineal AANAT is special--it responds to temperature in a species-specific manner, which reflects the fish ecophysiological preferences. This review summarizes anatomical, structural, and molecular aspects of the evolution of the melatonin-producing system in vertebrates.

Melatonin biosynthesis from serotonin involves the sequential activation of the arylalkylamine N-acetyltransferase (AANAT) and hydroxyindole-O-methyltransferase (HIOMT). Photoperiod synchronizes a daily rhythm in pineal and retinal melatonin secretion through controlling AANAT activity. Teleost fish possess two Aanat, one expressed in the retina (AANAT1) and the other expressed in the pineal gland (AANAT2). We report here the full-length cloning of Aanat1, Aanat2, SmHiomt and Otx5 (orthodenticle homeobox homolog 5) in the turbot (Scophthalmus maximus, Sm), a flatfish belonging to an evolutionary recent group of Teleost. The temporal expression pattern of the genes investigated is consistent with the idea that OTX5 is needed for photoreceptor specification, and that the pineal gland differentiates before the retina. SmAanat2 expression remained pineal specific during the period of time investigated, whereas SmOtx5 and SmHiomt expressions were seen in both the retina and pineal gland. Our results do not support the existence of a second SmHiomt, as is the case for SmAanat. Neither SmAanat2 nor SmHiomt mRNAs displayed cyclic accumulation in the pineal organ of embryos and larvae maintained under a light-dark cycle from fertilization onward. This is in marked contrast with the situation observed with zebrafish Aanat2, indicating that the molecular mechanisms controlling the development of the pineal melatonin system have been modified during the evolution of Teleost.

The cephalochordate amphioxus (Branchiostoma sp.) is an important animal model for studying the evolution of chordate developmental mechanisms. Obtaining amphioxus embryos is a key step for these studies. It has been shown that an increase of 3-4 degrees C in water temperature triggers spawning of the European amphioxus (Branchiostoma lanceolatum) in captivity, however, very little is known about the natural spawning behavior of this species in the field. In this work, we have followed the spawning behavior of the European amphioxus during two spawning seasons (2004 and 2005), both in the field and in captivity. We show that animals in the field spawn approximately from mid-May through early July, but depending on the year, they show different patterns of spawning. Thus, even if temperature has a critical role in the induction of the spawning in captivity, it is not the major factor in the field. Moreover, we report some improvements on the methodology for inducing spawning in captivity (e.g. in maintenance, light cycle control and induction of spawning in a laboratory without running sea water system). These studies have important implications for amphioxus animal husbandry and for improving laboratory techniques to develop amphioxus as an experimental animal model. J. Exp. Zool. (Mol. Dev. Evol.) 308B, 2007. (c) 2007 Wiley-Liss, Inc.

Melatonin is the time-keeping molecule of the organism. The production by the pineal organ is responsible for the diurnal and annual rhythms of plasma melatonin content. This contributes to synchronizing behavioural, biochemical and physiological processes to the environmental variations in photoperiod and temperature. Conservation and diversity characterize the melatonin system in vertebrates: conservation because its nocturnal pattern of production as well as its synchronizing properties are a constant; diversity because the modalities of its biosynthesis and modes of action have been profoundly modified in the course of evolution. This review summarizes our current knowledge on the targets and modes of action of melatonin in fish and comparisons are made with mammals.

International audience; Photoperiod plays an essential role in the synchronization of metabolism, physiology, and behavior to the cyclic variations of the environment. In vertebrates, information is relayed by the pineal cells and translated into the nocturnal production of melatonin. The duration of this signal corresponds to the duration of the night. In fish, the pinealocytes are true photoreceptors in which the amplitude of the nocturnal surge is modulated by temperature in a species-dependent manner. Thus, the daily and annual variations in the amplitude and duration of the nocturnal melatonin signal provide information on daily and calendar time. Both light and temperature act on the activity of the penultimate enzyme in the melatonin biosynthesis pathway, the arylalkylamine N-acetyltransferase (serotonin → N-acetylserotonin). Although the mechanisms of the light/dark regulation of melatonin secretion are quite well understood, those of temperature remain unelucidated. More generally, the mechanisms of thermoreception are unknown in ectotherms. Here we provide the first evidence that two thermotransient receptor potential (TRP) channels, TRPV1 and TRPV4, are expressed in the pineal photoreceptor cells of a teleost fish, in which they modulate melatonin secretion in vitro. The effects are temperature dependent, at least for TRPV1. Our data support the idea that the pineal of fish is involved in thermoregulation and that the pineal photoreceptors are also thermoreceptors. In other nervous and nonnervous tissues, TRPV1 and TRPV4 display a ubiquitous but quantitatively variable distribution. These results are a fundamental step in the elucidation of the mechanisms of temperature transduction in fish.

Melatonin (N-acetyl-5-methoxytrypamine) is the vertebrate hormone of the night: circulating levels at night are markedly higher than day levels. This increase is driven by precisely regulated increases in acetylation of serotonin in the pineal gland by arylalkylamine N-acetyltransferase (AANAT), the penultimate enzyme in the synthesis of melatonin. This unique essential role of AANAT in vertebrate timekeeping is recognized by the moniker the timezyme. AANAT is also found in the retina, where melatonin is thought to play a paracrine role. Here, we focused on the evolution of AANAT in early vertebrates. AANATs from Agnathans (lamprey) and Chondrichthyes (catshark and elephant shark) were cloned, and it was found that pineal glands and retinas from these groups express a form of AANAT that is compositionally, biochemically, and kinetically similar to AANATs found in bony vertebrates (VT-AANAT). Examination of the available genomes indicates that VT-AANAT is absent from other forms of life, including the Cephalochordate amphioxus. Phylogenetic analysis and evolutionary rate estimation indicate that VT-AANAT evolved from the nonvertebrate form of AANAT after the Cephalochordate-Vertebrate split over one-half billion years ago. The emergence of VT-AANAT apparently involved a dramatic acceleration of evolution that accompanied neofunctionalization after a duplication of the nonvertebrate AANAT gene. This scenario is consistent with the hypotheses that the advent of VT-AANAT contributed to the evolution of the pineal gland and lateral eyes from a common ancestral photodetector and that it was not a posthoc recruitment.

There is a need to understand NASA's Deep Space Network (DSN) coverage gaps and any limitations to provide redundant communication coverage for future deep space missions, especially for manned missions to Moon and Mars. The DSN antennas are required to provide continuous communication coverage for deep space flights, interplanetary missions, and deep space scientific observations. The DSN consists of ground antennas located at three sites: Goldstone in USA, Canberra in Australia, and Madrid in Spain. These locations are not separated by the exactly 120 degrees and some DSN antennas are located in the bowl-shaped mountainous terrain to shield against radiofrequency interference resulting in a coverage gap in the southern hemisphere for the current DSN architecture. To analyze the extent of this gap and other coverage limitations, simulations of the DSN architecture were performed. In addition to the physical properties of the DSN assets, the simulation incorporated communication forward link calculations and azimuth/elevation masks that constrain the effects of terrain for each DSN antenna. Analysis of the simulation data was performed to create coverage profiles with the receiver settings at a deep space altitudes ranging from 2 million to 10 million km and a spherical grid resolution of 0.25 degrees with respect to longitude and latitude. With the results of these simulations, two- and three-dimensional representations of the area without communication coverage and area with coverage were developed, showing the size and shape of the communication coverage gap projected in space. Also, the significance of this communication coverage gap is analyzed from the simulation data.

A methodology for inducing spawning in captivity of the lancelet Branchiostoma lanceolatum has been developed recently with animals collected at the Racou beach, in the southern coast of France. An increasing amount of laboratories around the world are now working on the evolution of developmental mechanisms (Evo-Devo) using amphioxus collected in this site. Thus, today, the development of new aquaculture techniques for keeping amphioxus in captivity is needed and the study of the natural conditions at which amphioxus is exposed in the Racou beach during their spawning season becomes necessary. We have investigated the amphioxus distribution, size frequency, and population structure in the Racou beach during its natural spawning season using multivariate methods (redundancy analysis and multiple regression). We found a clear preference of amphioxus for sandy sites, something that seems to be a general behaviour of different amphioxus species around the world. We have also estimated the amphioxus growth rate and we show how the animals are preferentially localized in shallow waters during April and June.

Members of the subphylum Cephalochordata, which include the genus Branchiostoma (i.e. amphioxus), represent the closest living invertebrate relatives of the vertebrates. To date, developmental studies have been carried out on three amphioxus species (the European Branchiostoma lanceolatum, the East Asian B. belcheri, and Floridian-Caribbean B. floridae). In most instances, adult animals have been collected from the field during their ripe season and allowed (or stimulated) to spawn in the laboratory. In any given year, dates of laboratory pawning have been limited by two factors. First, natural populations of these three most studied species of amphioxus are ripe, at most, for only a couple of months each year and, second, even when apparently ripe, animals spawn only at unpredictable intervals of every several days. This limited supply of living material hinders the development of amphioxus as a model system because this limitation makes it more difficult to work out protocols for new laboratory techniques. Therefore we are developing laboratory methods for increasing the number of amphioxus spawning dates per year. The present study found that a Mediterranean population of B. lanceolatum living near the Franco-Spanish border spawned naturally at the end of May and again at the end of June in 2003. Re-feeding experiments in the laboratory demonstrated that the gonads emptied at the end of May refilled with gametes by the end of June. We also found that animals with large gonads (both, obtained from the field and kept and fed at the laboratory during several weeks) could be induced to spawn in the laboratory out of phase with the field population if they were temperature shocked (spawning occurred 36 hours after a sustained increase in water temperature from 19°C to 25°C). J. Exp. Zool. (Mol. Dev. Evol.) 302B:384–391, 2004. © 2004 Wiley-Liss, Inc.

A female Octopus macropus, the first ever observed spawning, attached eggs (4·0×1·2 mm, with a chorion stalk 4 mm in length) individually or in clusters to a hard substrate, and brooded them till hatching. Hatchlings measured 4·0 mm in dorsal mantle length, 5·5 mm in total length and were planktonic; their short arms had seven suckers each, the outer demi-branch of each gill had ten lamellae. The chromatophore pattern confirmed an earlier identification of young individuals from the plankton (Naef, 1923).