The pH sensitivity of Aqp0 channels in tetraploid and diploid teleosts.

Water homeostasis and the structural integrity of the vertebrate lens is partially mediated by AQP0 channels. Emerging evidence indicates that external pH may be involved in channel gating. Here we show that a tetraploid teleost, the Atlantic salmon, retains 4 aqp0 genes (aqp0a1, -0a2, -0b1, and -0b2), which are highly, but not exclusively, expressed in the lens.

Mitochondrial aquaporin-8-mediated hydrogen peroxide transport is essential for teleost spermatozoon motility.

Reactive oxygen species (ROS), particularly hydrogen peroxide (H2O2), cause oxidative cell damage and inhibit sperm function. In most oviparous fishes that spawn in seawater (SW), spermatozoa may be exposed to harmful ROS loads associated with the hyperosmotic stress of axonemal activation and ATP synthesis from mitochondrial oxidative phosphorylation. However, it is not known how marine spermatozoa can cope with the increased ROS levels to maintain flagellar motility.

Development of a flatfish-specific enzyme-linked immunosorbent assay for Fsh using a recombinant chimeric gonadotropin.

In flatfishes with asynchronous and semicystic spermatogenesis, such as the Senegalese sole (Solea senegalensis), the specific roles of the pituitary gonadotropins during germ cell development, particularly of the follicle-stimulating hormone (Fsh), are still largely unknown in part due to the lack of homologous immunoassays for this hormone. In this study, an enzyme-linked immunosorbent assay (ELISA) for Senegalese sole Fsh was developed by generating a rabbit antiserum against a recombinant chimeric single-chain Fsh molecule (rFsh-C) produced by the yeast Pichia pastoris. The rFsh-C N- and C-termini were formed by the mature sole Fsh β subunit (Fshβ) and the chicken glycoprotein hormone common α subunit (CGA), respectively.

Flow cytometry analyses of phagocytic and respiratory burst activities and cytochemical characterization of leucocytes isolated from wrasse (Labrus bergylta A.).

We have isolated leucocytes from peripheral blood (PBL), head kidney (HKL) and spleen (SL) of wrasse (Labrus bergylta A.) and studied the innate immune responses phagocytosis and respiratory burst using flow cytometry. Further, we have characterized the phenotypic properties of the leucocytes by cytochemical staining.

Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication.

The granuloma disease caused by Francisella noatunensis subsp. noatunensis in farmed Atlantic cod has not been successfully treated by use of antibacterials, even when antibacterial resistance testing indicates a sufficient effect. The reason for this treatment failure may be the intracellular existence of the bacteria within immune cells, mainly macrophages.

Atlantic salmon (Salmo salar L.) smolts require more than two weeks to recover from acidic water and aluminium exposure.

The detrimental effects of acid rain and aluminium (Al) on salmonids have been extensively studied, yet knowledge about the extent and rate of potential recovery after exposures to acid and Al episodes is limited. Atlantic salmon smolts in freshwater (FW) were exposed for 2 and 7-day episodes (ACID2 and ACID7, respectively) to low pH (5.7±0.

Francisella noatunensis subsp. noatunensis replicates within Atlantic cod (Gadus morhua L.) leucocytes and inhibits respiratory burst activity.

Francisella noatunensis subsp. noatunensis, causing granulomatosis in cod, has been shown to reside within cod immune cells, mainly within monocytes and macrophages. In the present study, we analysed the ability of the bacterium to replicate within adherent cells isolated from head kidney by in vitro infection of leucocytes.

Short-term, but not long-term feed restriction causes differential expression of leptins in Atlantic salmon.

Atlantic salmon was used to investigate the effect of long- and short-term dietary ration on the tissue expression levels of leptins. Compared to ad libitum fed fish (0.8-3kg), 6months of dietary restriction (60%) resulted in significantly lower body mass and adiposity, but did not produce a clear effect on the expression levels of either lepa1 or lepa2.

Characterization of small, mononuclear blood cells from salmon having high phagocytic capacity and ability to differentiate into dendritic like cells.

Phagocytes are the principal component of the innate immune system, playing a key role in the clearance of foreign particles that include potential pathogens. In vertebrates, both neutrophils and mononuclear cells like monocytes, macrophages and dendritic cells are all professional phagocytes. In teleosts, B-lymphocytes also have potent phagocytic ability.

Effects of acidic water and aluminum exposure on gill Na(+), K(+)-ATPase alpha-subunit isoforms, enzyme activity, physiology and return rates in Atlantic salmon (Salmo salar L.).

Na(+), K(+)-ATPase (NKA) is involved, through its role as a major driving force for electrochemical gradients, in a range of transmembrane transport processes. Maintenance of homeostasis in anadromous salmonids requires modulation of several gill ion secretory proteins as part of the preparatory adaptation and acclimation to marine life. Atlantic salmon smolts were exposed to combinations of low pH and inorganic aluminum (acid/Al(i)) in freshwater (FW) and were then transferred to seawater (SW) for studies of post-smolt performance.

Oocyte selection is concurrent with meiosis resumption in the coenocystic oogenesis of Oikopleura.

Oogenesis in the tunicate, Oikopleura, is unusual for a chordate, in that the thousands of nuclei comprising the entire germline are contained in a unique giant cell, the coenocyst. We examined progression through meiotic prophase I in concert with cellular mechanisms implicated in selection, growth and maturation of oocytes in this shared cytoplasm. Unlike sister vertebrates, no germinal vesicle was formed and maternal transcripts were instead synthesized by polyploid nurse nuclei present in equal numbers to transcriptionally quiescent meiotic nuclei.

Enhancer detection and developmental expression of zebrafish sprouty1, a member of the fgf8 synexpression group.

Signaling pathways mediated by receptor tyrosine kinases (RTKs) are under positive and negative regulation, and misregulation of RTK signaling results in developmental defects and malignancy. A major class of antagonists of Fgf and Egf signaling are the Sprouty proteins. Through an enhancer detection approach, we isolated the sprouty1 (spry1) gene, expressed in multiple developing organs during embryogenesis.

Altered miRNA repertoire in the simplified chordate, Oikopleura dioica.

Recent studies reveal correlation between microRNA (miRNA) innovation and increased developmental complexity. This is exemplified by dramatic expansion of the miRNA inventory in vertebrates, a lineage where genome duplication has played a significant evolutionary role. Urochordates, the closest extant group to the vertebrates, exhibit an opposite trend to genome and morphological simplification.

Continued growth and cell proliferation into adulthood in the notochord of the appendicularian Oikopleura dioica.

The appendicularian urochordate Oikopleura dioica retains a free-swimming chordate body plan throughout life, in contrast to ascidian urochordates, whose metamorphosis to a sessile adult form involves the loss of chordate structures such as the notochord and dorsal nerve cord. Development to adult stages in Oikopleura involves a lengthening of the tail and notochord and an elaboration of the repertoire of tail movements. To investigate the cellular basis for this lengthening, we have used confocal microscopy and BrdU labeling to examine the development of the Oikopleura notochord from hatching through adult stages.

Endostyle cell recruitment as a frame of reference for development and growth in the Urochordate Oikopleura dioica.

In models of growth and life history, and in molecular and cell biology, there is a need for more accurate frames of reference to characterize developmental progression. In Caenorhabditis elegans, complete fate maps of cell lineage provide such a standard of reference. To be more widely applicable, reference frames should be easier to measure while still providing strong predictive capacity.

Spatiotemporal patterns of neurogenesis in the appendicularian Oikopleura dioica.

Incorporation of the thymidine analog bromodeoxyuridine (BrdU) was used to assess cytogenesis in the central nervous system (CNS) of the appendicularian Oikopleura dioica. A series of timed cumulative labelings carried out from 45 minutes (min) to 8 hours (h) after fertilization provided labeling patterns that showed when neurons and support cells residing at specific sites within the 9 h CNS became postmitotic. Throughout the CNS, which includes the cerebral ganglion, caudal ganglion and caudal nerve cord, neurogenesis occurs during an earlier time window than the genesis of support cells.

Endocrine systems in juvenile anadromous and landlocked Atlantic salmon (Salmo salar): seasonal development and seawater acclimation.

The present study compares developmental changes in plasma levels of growth hormone (GH), insulin-like growth factor I (IGF-I) and cortisol, and mRNA levels of their receptors and the prolactin receptor (PRLR) in the gill of anadromous and landlocked Atlantic salmon during the spring parr-smolt transformation (smoltification) period and following four days and one month seawater (SW) acclimation. Plasma GH and gill GH receptor (GHR) mRNA levels increased continuously during the spring smoltification period in the anadromous, but not in landlocked salmon. There were no differences in plasma IGF-I levels between strains, or any increase during smoltification.

Development of the caudal nerve cord, motoneurons, and muscle innervation in the appendicularian urochordate Oikopleura dioica.

The development of the caudal nerve cord and muscle innervation in the appendicularian Oikopleura dioica was assessed using differential interference contrast and confocal microscopy, phalloidin staining of actin, and in situ hybridization for the neuronal markers tubulin and choline acetyltransferase (ChAT). The caudal nerve cord first appears as a stream of tubulin mRNA-positive neurons that extends into the tail from the caudal ganglion. By this stage a few actin-rich nerve fibers course longitudinally along the cord.

Rapidly evolving lamins in a chordate, Oikopleura dioica, with unusual nuclear architecture.

Metazoan lamins are implicated in the organization of numerous critical nuclear processes. Among chordates, the appendicularian, Oikopleura dioica, has an unusually short life cycle involving rapid growth through extensive recourse to endoreduplication, a characteristic more associated with some invertebrates. In some tissues, this is accompanied by the formation of elaborate, bilaterally symmetric nuclear morphologies associated with specific gene expression patterns.

Phosphorylation of the histone H3.3 variant in mitosis and meiosis of the urochordate Oikopleura dioica.

Mammalian histone variant H3.3 differs from replication-dependent histone H3.1 by five amino acids, including replacement of alanine 31 by serine.

Conserved patterns of nuclear compartmentalization are not observed in the chordate Oikopleura.

Background Information: Recent results from a limited number of eukaryotic model organisms suggest that major principles governing spatial organization of the genome in functionally distinct nuclear compartments are conserved through evolution.

Results: We examined the in situ spatial organization of major nuclear components and nuclear patterns of gene loci with strictly defined expression patterns in endocycling cells of the transparent urochordate Oikopleura dioica, a complex metazoan with a very compact genome. Endocycling cells with different functions and similar DNA content displayed distinct topologies of nuclear components.

The Oikopleura coenocyst, a unique chordate germ cell permitting rapid, extensive modulation of oocyte production.

The ability to adjust reproductive output to environmental conditions is important to the fitness of a species. The semelparous, chordate, Oikopleura dioica, is particularly adept in producing a highly variable number of oocytes in its short life cycle. Here we show that this entails an original reproductive strategy in which the entire female germline is contained in a single multinucleate cell, the "coenocyst".

The cytoskeleton organizes germ nuclei with divergent fates and asynchronous cycles in a common cytoplasm during oogenesis in the chordate Oikopleura.

Germline cysts are conserved structures in which cells initiating meiosis are interconnected by ring canals. In many species, the cyst phase is of limited duration, but the chordate, Oikopleura, maintains it throughout prophase I as a unique cell, the coenocyst. We show that despite sharing one common cytoplasm with meiotic and nurse nuclei evenly distributed in a 1:1 ratio, both entry into meiosis and subsequent endocycles of nurse nuclei were asynchronous.

Comparative organization of follicle, accessory cells and spawning anlagen in dynamic semelparous clutch manipulators, the urochordate Oikopleuridae.

Background Information: The urochordate appendicularians play a key trophic role in marine ecosystems and are the second largest component of zooplankton after copepods. Part of their success is due to their ability to undergo rapid population blooms in response to changes in primary productivity. Nonetheless, the reproductive biology of this important group remains poorly understood.

Development of putative GABAergic neurons in the appendicularian urochordate Oikopleura dioica.

Studying the developing brain of urochordates can increase our understanding of brain evolution in the chordate lineage. To begin addressing regional patterns of neuronal differentiation in appendicularian urochordates, we examined the development of putative GABAergic neurons in Oikopleura dioica using GABA immunohistochemistry and in situ hybridization for the GABA-synthesizing enzyme GAD. First, we assessed the developmental dynamics of neuron number and organization in the cerebral and caudal ganglia.