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Cholinergic and adrenergic influence on the teleost heart in vivo.

The tonical cholinergic and adrenergic influence on the heart rate was investigated in vivo in seven species of marine teleosts (pollack, Pollachius pollachius; cuckoo wrasse, Labrus mixtus; ballan wrasse, Labrus berggylta; five-bearded rockling, Ciliata mustela; tadpole fish, Raniceps raninus; eel-pout, Zoarces viviparus and short-spined sea scorpion, Myoxocephalus scor pius) during rest and, in two of the species (P. pollachius and L. mixtus), also during moderate swimming exercise in a Blazka-type swim tunnel. Ventral aortic blood pressure and heart rate were recorded via a catheter implanted in an afferent branchial artery, and the influence of the cholinergic and adrenergic tonus on the heart rate was assessed by injection of atropine and sotalol respectively. During rest the adrenergic tonus was higher than the cholinergic tonus in all species except L. berggylta, where the reverse was true. In P. pollachius and L. mixtus, exercise appeared to produce a lowering of the cholinergic tonus on the heart and, possibly, a slight increase of the adrenergic tonus. The nature of the adrenergic tonus (humoral or neural) is not clear, but the low plasma concentrations of catecholamines both during rest and exercise could be interpreted in favour of a mainly neural adrenergic tonus on the teleost heart. These experiments are compatible with the view that both a cholinergic inhibitory tonus and an adrenergic excitatory tonus are general features in the control of the teleost heart in vivo, both at rest and during moderate swimming exercise.

Animals↗

[Characteristics of the conditioned motor reactions of unrestrained elasmobranches and teleosts].

In elasmobranch (Scyllium canicula, Galleus canis) and teleost (Migull capitocum) fishes it is possible to form motor food-searching conditioned reflexes to discrimination of light and darkness. Some differences were revealed in ecological and conditioned motor behavioural activities in higher and lower sharks. Elasmobranch and teleost fishes exhibit significant differences in ecological, feeding and conditioned reflex behaviour. Nervous activity in elasmobranchs is characterized by lower and primitive organization as compared to that in teleosts.

Animals↗

Ultrastructural studies on the atrio-ventricular junction and ventriculo-bulbar junction in heart of teleosts.

The ultrastructure of the atrio-ventricular junction (A-V junction) and ventriculo-bulbar junction in the heart of a teleostean species, Gadus virens L., is described. The former junction is a narrow canal having a length of 0.35 to 0.60 mm and a wall thickness of 0.20 to 0.32 mm. At the lumenal side of the junctional wall occurs a layer of myocardial tissue which is in continuity with that of the atrium and ventricle. Further, at the atrial side of the junction this tissue is trabeculated, whereas it is untrabeculated at the ventricular side. A number of nerves occur near the atrium. The myocardial cells are tightly packed into bundles, which are surrounded by a thick external lamina. These cells are connected by numerous nexuses and desmosomes, and are rich in myofibrils. At the atrial side of the A-V junction the endocardial cells contain numerous bristle-coated vesicles, tubules of agranular endoplasmic reticulum and large inclusion bodies of moderate electron density, whereas at the ventricular side these cells are poor in bristle-coated vesicles and agranular endoplasmic reticulum, and the inclusion bodies are small and electron dense. The ventriculo-bulbar junction is a narrow canal; the wall is 0.5 to 1.0 mm thick and contains outermost high amounts of loose collagen tissue, and innermost a layer of dense collagen tissue. Between the 2 latter layers occurs a layer of myocardial tissue which is continuous with that of the ventricle. The endocardial cells in this junction is structurally similar to those located at the ventricular side of the A-V junction. The structure of the A-V junction in teleosts is compared with that of the A-V node and bundle of His in mammals, and the sino-atrial junction in teleosts. It is proposed that the myocardial tissue of the A-V junction in teleosts may conduct electrical impulses between atrium and ventricle.

Animals↗

Physical biochemical properties of IgM from a teleost fish.

Teleost fish are able to produce IgM class antibody, as are other vertebrates. When the teleost fish Oreochromis niloticus was immunized with bovine serum albumin (BSA), it produced antibody to BSA with an average avidity of 7.4 x 10(8)/M. Thus, dissociation of antigen-antibody complexes only occurred at conditions of < pH 2.5, > pH 11, > 4M NaI or > 4M urea, demonstrating high stability of the complex. Western blot analyses further showed the high specificity of the antibody to BSA. In contrast to mammals, when the fish was challenged with multiple protein antigens, it produced antibody only to the major component but not to others. The antibody generated to a specific antigen accounted for up to 1.1% of whole serum protein or 7.0% of whole immunoglobulin. We conclude (1) the systemic antibody response in teleost fish may be an 'all or nothing' response, which is different from that in mammals; (2) the quality (specificity and affinity) of the antibody produced is similar to that of mammals. The findings not only reveal a quite different strategy of immune response in fish, but also raise the possibility of technical application.

Animals↗

The morphology of teleost meningocytes as revealed by freeze-fracture.

We describe the morphology of the cells in the three layers of the teleost endomeninx, as viewed by freeze-fracturing. The cells of the outer endomeningeal layer are fusiform and closely packed, with interdigitations that hold the cells together, gap junctions and a few strands of particles resembling tight junctions, but no desmosomes. The intermediate layer is formed by a single layer of flattened and elongated cells with rectangular shape, and well developed junctional complexes (gap junctions, tight junctions and desmosomes). These cells also show numerous plasmalemmal vesicles (6.5 +/- 1.3/microns 2) in the upper (in contact with the outer layer) and lower (in contact with the inner layer) membranes. Cross fracture of these cells shows many membrane-bound and free vesicles. The inner layer is formed by spindle shaped cells with wide intercellular spaces filled with a granular matrix and collagen fibers. The density of intramembrane particles is higher than that in the meningocytes of the outer and intermediate layers. The morphology of the teleost endomeninx appears similar to that of the leptomeninges of birds and mammals, but the outer endomeningeal layer of teleosts (which resembles the arachnoid of elasmobranchs and amphibians) appears different from any cell layer in the meninges of amniotic vertebrates.

Animals↗

Structure and diversity of the TCR alpha-chain in a teleost fish.

T cell receptor beta-chain genes are well characterized in representatives of most vertebrate phyla, from sharks to mammals, but the molecular structure of complete TCR alpha-chains has not yet been established in cold-blooded vertebrates. We used a PCR approach to isolate cDNAs encoding putative teleost fish (Oncorhynchus mykiss, rainbow trout) TCR alpha-chains. Eight V alpha segments were identified, belonging to six different families, and the best amino acid sequence identity scores for these trout V alpha were all provided by mammalian V alpha or V delta sequences. Twenty-four (60.1 %) of the 39 analyzed V alpha segments belong to the V alpha 2 family, which has limited homology with mammalian V alpha/delta sequences and with the human V pre-B sequence. A total of 32 different J alpha segments were identified from 40 J alpha regions sequenced, suggesting that a large repertoire of J alpha segments is a characteristic of most vertebrates. The structural properties of the TCR alpha-chain complementarity-determining region 3 loop are well conserved between trout and mammals, suggesting that this region has been under continuous selective pressure in jawed vertebrate evolution. The trout C alpha segment has conserved N-terminal and transmembrane domains, but the C alpha intercysteine distance contains only 40 residues, significantly smaller as compared with mammals (49-56 residues). The conserved features of teleost fish TCR beta- and alpha-chains with their mammalian equivalents suggest that TCR-alpha beta receptors were still present in the common Devonian ancestors of modern teleost fish and mammals, about 450 million years ago.

Amino Acid Sequence↗

[The ratio of processes of lipid peroxidation and antioxidant activity in gonads of Elasmobranchia and teleost fishes of the Black Sea].

It has been found that gonads of sharks contain higher content of alkoxylipids and vitamin K and lower levels of monoglycerides and carotenoids than gonads of teleost fishes. The peroxidase activity is higher in gonads of shark males but the glutathione reductase activity is lower. No significant differences in lipids peroxidation indices in gonads of elasmobranchia and teleosts were found. The total antioxidant activity of lipids in gonads of shark males exceeds 1, while the same indices of teleosts are below 1. Differences in the lipid composition, antioxidant enzyme activities, the content of glutathione, carotenoids, vitamins A, E, K and lipid peroxidation in male and female fish gonads are described. Correlation coefficients for all indices of the lipid peroxidation and antioxidant activity are estimated.

Animals↗

McAB 300 antibody against calbindin D-28K is a glial marker in the teleost brain.

The monoclonal antibody McAB 300 against calbindin D-28 k demonstrated an immunostaining pattern in the teleost brain completely different to those obtained in other classes of vertebrates, or after using in the teleost brain other monoclonal and polyclonal antibodies raised against the same protein. Although calbindin D-28k is considered a neuronal marker, McAB 300 specifically stained glial cells throughout the brains of teleosts, with the only exception of some retinal amacrine and horizontal cells.

Animals↗

Teleost lincRNAs: Functional roles, regulatory mechanisms, and future applications in aquaculture.

Long intergenic non-coding RNAs (lincRNAs) regulate gene expression across vertebrate physiological systems, yet their functional roles in teleost fish remain incompletely synthesized. This review systematically integrates current evidence through PRISMA-guided searches across PubMed, Web of Science, and Scopus, identifying ten lincRNA-focused functional studies with genetic, mechanistic, or developmental validation, complemented by twenty two supplementary contextual references. Findings span development, immunity, environmental adaptation, reproduction, regeneration, and toxicology, with each association graded as experimentally validated, bioinformatically predicted, correlational, or speculative. This synthesis offers three core contributions. First, it shows that cis-acting regulation on neighboring genes, mediated through Wnt, NF-&#x3ba;B, and AHR signaling, is the dominant validated lincRNA mechanism across teleost physiological domains. Second, it demonstrates that direct experimental validation, primarily via CRISPR-Cas9 and chromatin-capture assays, remains concentrated in zebrafish, whereas aquaculture-species associations remain largely correlational. Third, it identifies two findings that challenge current lincRNA classification: unexpected regulatory directionality at the slincR-sox9b locus, and micropeptide-encoding potential within annotated lincRNAs. Together, these contributions establish an evidence-graded foundation for future mechanistic studies and translational aquaculture applications.

Aquaculture↗

DNA methylation reprogramming in teleosts.

Early embryonic development is crucially important but also remarkably diverse among animal taxa. Axis formation and cell lineage specification occur due to both spatial and temporal control of gene expression. This complex system involves various signaling pathways and developmental genes such as transcription factors as well as other molecular interactants that maintain cellular states, including several types of epigenetic marks. 5mC DNA methylation, the chemical modification of cytosines in eukaryotes, represents one such mark. By influencing the compaction of chromatin (a high-order DNA structure), DNA methylation can either repress or induce transcriptional activity. Mammals exhibit a reprogramming of DNA methylation from the parental genomes in the zygote following fertilization, and later in primordial germ cells (PGCs). Whether these periods of methylation reprogramming are evolutionarily conserved, or an innovation in mammals, is an emerging question. Looking into these processes in other vertebrate lineages is thus important, and teleost fish, with their extensive species richness, phenotypic diversity, and multiple rounds of whole genome duplication, provide the perfect research playground for answering such a question. This review aims to present a concise state of the art of DNA methylation reprogramming in early development in fish by summarizing findings from different research groups investigating methylation reprogramming patterns in teleosts, while keeping in mind the ramifications of the methodology used, then comparing those patterns to reprogramming patterns in mammals.

Animals↗

Visceral afferent and efferent columns in the spinal cord of the teleost, Ictalurus punctatus.

In tetrapod vertebrates, neural circuitries subserving visceral and somatic reflexes are each represented in distinct columns of cells within the gray area of the spinal cord. To determine the location of visceral elements of the spinal cord of a teleost fish, crystals of the carbocyanine dye 1,1'dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine (DiI), were placed on either the abdominal sympathetic (mesenteric) nerves, the coeliac ganglia, or on the rostral three somatic spinal nerves, in fixed specimens of the channel catfish, Ictalurus punctatus. In fish in which DiI had been placed on the mesenteric nerves, labeled fibers coursed along the lateral margin of the dorsal horn within the first and second spinal segments, and appeared to terminate in a region at the base of the dorsal horn. In contrast, when DiI crystals were placed on the somatic spinal nerves, labeled primary afferents terminated in the dorsalmost two thirds of the dorsal horn, as well as in ventral and ventromedial areas of the medial funicular nuclear complex. Labeled somata (motor neurons) were situated in the ventral horn. When DiI crystals were placed bilaterally on the coeliac ganglia, labeled piriform and fusiform preganglionic neurons occurred in intermediate positions adjacent to the central canal, corresponding to the paracentral nucleus of Herrick, and in the lateral funiculus. These results demonstrate that somatic and visceral afferent and efferent functional columns are distinct in a teleost fish as they are in amniote vertebrates.

Animals↗

Effects of glutamic acid and related agents on horizontal cells in a marine teleost retina.

Excitatory amino acids (EAAs) such as glutamic and aspartic acids, considered as the most likely neurotransmitters at the photoreceptor-horizontal cell synapse of teleost retinas, as well as agonists such as kainic acid and several of their antagonists, were applied to isolated and superfused retinas of the teleost Eugerres plumieri. Intracellular recordings from horizontal cells reveal that EAA receptors are of the kainate-quisqualate type. There is competitive inhibition between the agonist and antagonist agents used, and under their combined effect, the synapse under study remains operational, in a functional state, able to modulate the horizontal cell membrane potential upon retinal illumination.

Animals↗

Origins of horizontal cell spectral responses in the retina of marine teleosts (Centropomus and Mugil sp).

Intracellular recording and marking of horizontal cells (HCs) were carried out in isolated retinas of marine teleosts (Centropomus and Mugil sp.) to identify the cellular origins of different spectral types of S-potential. The spectral responses recorded under mesopic conditions were classified into four types: photopic L, biphasic C(R/G), biphasic C(G/R), and scotopic L(sL). Intracellular marking with Lucifer yellow (LY) or Procion yellow (PY) revealed that in the Centropomus retina an L-type response was recorded from H1 and H2 cells, C(R/G)-type from H3 cells, and sL-type from H4 cells. However, in 20% of microelectrode penetrations from the photoreceptor surface, the sequential order of response appearances was found to be L-, sL-, and C(R/G)-types. In this species, thick dendritic processes of H3 and H4 cells are distributed at the same level as the cell body. In the Mugil retina, on the other hand, L-type response was recorded from H1 cells, C(G/R)-type from H2 cells, C(R/G)-type from H3 cells, and sL-type from H4 cells, this sequential arrangement being very regular. In both species, the cone-connected H1 cells are small and possess an axon, while the two other cone HCs as well as rod-connected H4 cells are axonless. Earlier (Negishi et al., 1988) and present findings indicate that there is a wide variety of HC morphology and functional organization in the teleost retina.

Animals↗

Nasotemporal asymmetry during teleost retinal growth: preserving an area of specialization.

Teleost fish retinas grow throughout adult life through both cell addition and stretching. Cell division occurs at the periphery of the retina, resulting in annular addition of all cell types except rod photoreceptors, which are added in the central retina. Since many teleosts have a region of high cellular density at the temporal pole of the eye, we analyzed whether and how this specialized region of high visual acuity maintained its relative topographical position through asymmetric circumferential growth. To do this, we measured the pattern of long-term retinal growth in the African cichlid Haplochromis burtoni. We found that the retina expands asymmetrically along the nasotemporal axis, with the nasal retina growing at a higher rate than the temporal, dorsal, or ventral retinae, whose growth rates are equal. This nasotemporal asymmetry is produced via significantly greater expansion of retinal tissue at the nasal pole rather than through differential cell proliferation. The mechanisms responsible for this differential retinal enlargement are unknown; however, such asymmetric expansion very likely minimizes disruption in vision during rapid growth.

Animals↗

Postovulatory follicle: a model for experimental studies of programmed cell death or apoptosis in teleosts.

This is the first evidence of programmed cell death, or apoptosis, occurring in the postovulatory follicle (POF) of teleost fish. Females of Astyanax bimaculatus lacustris were submitted to induced ovulation through injecting pituitary extract. Ultrastructural analyses of POFs at time intervals varying from zero to four days postspawning showed several characteristic events of the apoptosis. Typical apoptotic figures, such as nucleus with chromatin condensation underlying the nuclear envelope in a crescent pattern and apoptotic bodies at different stages of formation and reabsorption, were observed in the follicular cells a few days after the onset of the postovulatory period. The results indicated that apoptosis is the major mechanism responsible for the elimination of the follicular cells in the POFs of A. bimaculatus lacustris during ovarian recovery postspawning. It is suggested that POFs might be used as an experimental model in dynamic studies involving cell death in teleosts.

Animals↗

Flow cytometry-based assay for determination of teleost cytotoxic cell lysis of target cells.

BACKGROUND: The nonradiometric assays previously developed to detect cellular cytotoxic activity have been hindered by many difficulties. Among the problems are the requirement for expensive commercial kits and the use of techniques that produce high background noise and decreased sensitivity. In addition, these assays did not account for bidirectional apoptosis (activation-induced cell death [AICD]). Most attempts to derive cytometry-based cytotoxicity assays have been unsuccessful because individual effectors and targets could not be identified (i.e., "separated") using gating techniques. METHODS: In the present study, teleost nonspecific cytotoxic (NCC) and mammalian target cells were each sufficiently different in size to identify them by flow cytometry (FCM). Using appropriate gating and discriminator techniques, these two cell populations were differentiated based on scatter properties and propidium iodide (PI) binding. Total capacity for PI binding was obtained by permeabilization of the targets with ice-cold acetone. Spontaneous PI binding was relatively low. This technique detected cytotoxicity at effector-to-target ratios (E:T) of 1:1 and after only 30 min cocultivation. RESULTS: Tilapia NCC from peripheral blood kill human transformed target cells by necrosis and apoptosis as identified by PI binding. Maximum killing of HL-60 targets (approximately 100%) occurred by 180 min cocultivation. For the same time, the killing of IM-9 did not exceed 60%. Almost 90% of IM-9 targets are lysed following 14 h of cocultivation. The maximum killing of both HL-60 and IM-9 targets was observed at a 25:1 E:T ratio after 14 h. Comparisons of the chromium(>51) release assay with flow detection of cytotoxicity revealed that FCM detected 55% lysis of the target cells compared with 2% cytotoxicity by chromium release, after a cocultivation time of 240 min. DISCUSSION: FCM detection of (teleost) NCC lysis of target cells using PI uptake is more sensitive than standard chromium release assays. This level of sensitivity was observed because NCC and targets were sufficiently different in size such that they could be resolved by scatter plots. Using FCM, cytotoxicity was detected earlier and at lower E:T ratios than previously reported for chromium release assays. Although tilapia were reported previously to be not capable of lysing IM-9 targets by chromium release detection, the more sensitive method of FCM detected cytotoxicity using PI uptake. HL-60 lysis by tilapia NCC exhibited saturable kinetics but occurred at different times post-cocultivation.

Animals↗

Identification, characterization, and ontogeny of a second cytochrome P450 3A gene from the fresh water teleost medaka (Oryzias latipes).

Multiple copies of cytochrome P450 gene family 3 have been identified from numerous mammalian species. Often these genes exhibit differential catalytic activities and gene regulation. To date however, little information is available regarding multiple forms of this gene family in teleost fishes. In this study, a second isozyme of cytochrome P450 3A has been cloned from the teleost fish Oryzias latipes and designated CYP3A40. Screening of a cDNA library to medaka liver resulted in the identification of a full length cDNA clone containing a 2316 base pair (bp) insert with an open reading frame encoding a single peptide of 502 amino acids. Comparisons of the deduced amino acid sequence to other known cytochrome P450 sequences indicate that this gene product is most similar to the CYP3A gene family and shares a 90% identity to CYP3A38 previously identified from medaka liver. Consistent with Northern blot and Western blot analysis, Southern blots of medaka genomic DNA demonstrated the presence of two CYP3A genes. Gene expression studies demonstrated that CYP3A38 and CYP3A40 are differentially regulated according to embryonic development. Northern blot analysis, using a probe to a conserved region of both CYP3A genes, demonstrated the presence of a single CYP3A transcript for early and late embryonic stages and two CYP3A transcripts in larvae and adult liver. Similarly, Western blots show a single faint immunoreactive cytochrome P450 3A protein in microsomes from early and late embryos and two abundant protein bands in microsomes from larval and adult liver. To further examine the transcriptional differences in CYP3A expression, RT-PCR analysis was performed on embryonic stages 11-35, 1- and 14-day-old larvae, and adult liver using primer sets specific for CYP3A38 and CYP3A40. These results demonstrate that CYP3A40 is expressed early in embryonic development and continues throughout adult stages. CYP3A38, however, is tightly regulated during embryonic development and is only expressed post-hatch.

Amino Acid Sequence↗

Search for enhancers: teleost models in comparative genomic and transgenic analysis of cis regulatory elements.

Homology searches between DNA sequences of evolutionary distant species (phylogenetic footprinting) offer a fast detection method for regulatory sequences. Because of the small size of their genomes, tetraodontid species such as the Japanese pufferfish and green spotted pufferfish have become attractive models for comparative genomics. A disadvantage of the tetraodontid species is, however, that they cannot be bred and manipulated routinely under laboratory conditions, so these species are less attractive for developmental and genetic analysis. In contrast, an increasing arsenal of transgene techniques with the developmental model species zebrafish and medaka are being used for functional analysis of cis regulatory sequences. The main disadvantage is the much larger genome. While comparison between many loci proved the suitability of phylogenetic footprinting using fish and mammalian sequences, fast rate of change in enhancer structure and gene duplication within teleosts may obscure detection of homologies. Here we discuss the contribution and potentials provided by different teleost models for the detection and functional analysis of conserved cis-regulatory elements.

Animals↗