Peptides related to insulin-like growth factor 1 in the gastro-entero-pancreatic system of bony and cartilaginous fish.

Evidence for the presence of peptides, related to insulin-like growth factor 1 (IGF-1) has been obtained in serum and various organs of representatives of osteichthyes and chondrichthyes, i.e., the bony fish Myoxocephalus (Cottus) scorpius and the cartilaginous fish Raja clavata. The peptides were identified by means of gel chromatography and an IGF-1 radioimmunoassay. IGF-1-like immunoreactivity was detected in three different apparent molecular mass forms, i.e., 17 kDa, 6 kDa and 4 kDa, the occurrence of which seemed to depend on the species. When the same antiserum was used immunohistochemically, IGF-1-like immunoreactivity was observed in endocrine cells of the open type in the intestinal mucosal epithelium. These cells exhibited distinct and species-specific distribution patterns. Endocrine cells of the pancreas as well as epithelial cells of the pancreatic duct also showed IGF-1-like immunoreactivity. Occasionally, IGF-1-like immunoreactivity was observed also in interstitial cells. The distribution patterns and densities of the IGF-like immunoreactive cells correlated with the results obtained by radioimmunoassay of the crude extracts. Absorption studies indicated that the IGF-1-like peptides observed differ from mammalian and submammalian insulins as well as from mammalian IGF-1.     

Genomics and mapping of teleostei (bony fish)

Until recently, the Human Genome Project held centre stage in the press releases concerning sequencing programmes. However, in October 2001, it was announced that the Japanese puffer fish (Takifugu rubripes, Fugu) was the second vertebrate organism to be sequenced to draft quality. Briefly, the spotlight was on fish genomes. There are currently two other fish species undergoing intensive sequencing, the green spotted puffer fish (Tetraodon nigroviridis) and the zebrafish (Danio rerio). But this trio are, in many ways, atypical representations of the current state of fish genomic research. The aim of this brief review is to demonstrate the complexity of fish as a group of vertebrates and to publicize the 'lesser-known' species, all of which have something to offer.     

Comparative analysis of gastrin-immunoreactive and argyrophil cells in bony fish larvae

A comparative study was made of enteroendocrine cells in the larvae of bony fishes (Teleostei): carp (Cyprinus carpio L.), big head (Aristichyts nobilis Richardson), silver carp (Hypophthalmichthys molitrix Valenciennes), and atlantic salmon (salmo salar L.). Argyrophil cells demonstrated by the method after Grimelius were found in all examined fish species. They are located in the pyloric appendices, stomach and intestinal mucosa. Gastrin-immunoreactive cells demonstrated by the immunocytochemical method were located in the pyloric part of the stomach in all examined fishes, except for the atlantic salmon in which they were not identified. The first argyrophil cells were identified at the age of about 10 days, while gastrin-immunoreactive cells proved to appear 5-7 days later.     

Conformational comparisons among homologous bony fish 5S ribosomal RNAs

The reversible thermal melting of homologous 5S ribosomal RNAs of three bony fishes, rainbow trout (Salmo gairdneri), carp (Cyprinus carpio), and dog salmon (Onchorhynchus keta), were compared in aqueous salts (sodium chloride and/or divalent metal chlorides) by means of differential optical absorption at 260 and 280 nm, and c.d. at 265 nm. The melting parameters of the multiphasic melting process were conserved among the homologous RNAs, and it was shown that they did not depend on the methods of measurement but on concentrations of [Na+] and of [metal2+]. Other physicochemical methods, such as FT-IR absorption, gave the same melting profiles as the u.v. methods. The c.d. spectrum was homologous among the ribosomal RNAs. The melting is explained by changes in local structures in the 'consensus secondary structure model'. Also, stabilization of the secondary and tertiary structures by binding of Mg2+ and Ca2+, and the catalysed hydrolysis of the main chains by various divalent ions are discussed.     

Multiple sex-chromosome system in a loach fish

This study shows an X1X1X2X2/X1X2Y type of multiple sex-chromosome system in the genus Cobitis (Pisces, Ostariophysi). Observation of C-banded male meioses revealed that one arm of the neo-Y always associates end-to-end with one of the X chromosomes. This finding implies that a mechanism might be present that has prevented crossing-over between the ancestral X and Y chromosomes without morphological differentiation. This is the first reported case of multiple sex chromosomes among cyprinoid fish.     

Universal mtDNA primers for species identification of degraded bony fish samples

We developed primers for amplifying and sequencing highly degraded mtDNA from diverse fish species. The primers flank a variable 148-bp fragment within the 12S region of mtDNA. We screened and sequenced 82 samples of bony fishes representing 17 families to confirm cross-species amplification and identification. Salmonid species were analysed and demonstrate 13 species-specific SNPs within this region. Based on alignments of additional deposited sequences, these primers are conserved in many other species, making them useful for species identification using degraded DNA samples such as archaeological specimens.     

Comparative macroscopic and microscopic anatomical observations on pyloric ceca in some bony fish

The authors report the variable number of pyloric caeca of some species of fish and describe their macroanatomy. They also describe and compare the microanatomy of pyloric caeca in three species; in particular they point out the thickness of tunica muscularis, the shape and length of villi which protrude into the lumen and the variable number of goblet cells. The tunica muscularis is very strong in Uranoscopus and consists of three strata: external and inner longitudinal strata, middle circular stratum. The tunica muscularis is less thick in Diplodus and in Scorpaena. The lamina propria and epithelium form villi which protrude into the lumen. They are covered with columnar epithelium and goblet cells which are more numerous in Uranoscopus and Scorpaena and less numerous in Diplodus. At last the authors describe in pyloric caeca of Scorpaena the presence of a network which engages all the lumen; it consists of connective tissue septa which directly derive from the lamina propria and is covered with the same columnar epithelium and goblet cells.     

Investigating the morphology, function and genetics of cytotoxic cells in bony fish

Bony fish (teleosts) possess multiple cytotoxic cell lineages that recognize and destroy virally infected and transformed cells. In general, these lineages parallel their functional equivalents in mammals and include neutrophilic granulocytes, macrophages, cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. These four cell types have been morphologically identified in multiple fish species but only limited information is available about their function. In contrast, much work has gone into examining the function of a fifth cytotoxic cell lineage, termed nonspecific cytotoxic cells (NCC), that has been referred to as the bony fish equivalent of NK cells. However, evidence suggesting that NCC do not represent the NK lineage has come through the development of multiple cytotoxic catfish cell lines that are morphologically and functionally similar to human NK cells and are distinct from NCC. In addition to characterizing cytotoxic cells from fish, recent work has identified the novel immune-type receptors (NITR) and cichlid killer leukocyte receptors (cKLR) that are structurally related to mammalian NK receptors and likely play a role in cytotoxic function in fish. This review summarizes the morphological and functional evidence for cytotoxic cells within bony fish and discusses future directions for examining cytotoxicity through genomics and transgenics.     

Parathyroid hormone-related protein (PTHrP) in cartilaginous and bony fish tissues.

Tissues from a range of fish were examined for the presence of parathyroid hormone-related protein (PTHrP) to investigate PTHrP protein distribution and PTHrP gene expression in jawless fish, cartilaginous fish, and bony fish. Immunoreactive PTHrP was localized using antisera to N-terminal and mid-molecule regions of human PTHrP and PTHrP gene expression examined using a digoxigenin labeled riboprobe to a conserved region of the mammalian PTHrP gene. In all of the fish studied, PTHrP protein and messenger RNA (mRNA) were localized to the skin, kidney, and skeletal muscle, following the pattern seen in higher vertebrates. Additional sites of localization for both protein and mRNA included gill, nerve cord, and pituitary, as well as developing dermal denticles and rectal gland in the elasmobranch species. The sites of PTHrP distribution indicate that PTHrP may have roles in ionoregulation as well as growth and differentiation in fish, as has been suggested in higher vertebrates. The results imply that the distribution of PTHrP is widespread in fish and that there is homology between the PTHrP molecules found in humans and fish. The conservation of localization and possible similarity of the PTHrP molecules between tetrapods and fish suggests that PTHrP has a number of fundamental roles in vertebrates. J. Exp. Zool. 284:541-548, 1999.     

Nonlinear acoustic complexity in a fish 'two-voice' system

Acoustic signals play essential roles in social communication and show a strong selection for novel morphologies leading to increased call complexity in many taxa. Among vertebrates, repeated innovations in the larynges of frogs and mammals and the syrinx of songbirds have enhanced the spectro-temporal content, and hence the diversity of vocalizations. This acoustic diversification includes nonlinear characteristics that expand frequency profiles beyond the traditional categorization of harmonic and broadband calls. Fishes have remained a notable exception to evidence for such acoustic innovations among vertebrates, despite their being the largest group of living vertebrates that also exhibit widespread evolution of sound production. Here, we combine rigorous acoustic and mathematical analyses with experimental silencing of the vocal motor system to show how a novel swim bladder mechanism in a toadfish enables it to generate calls exhibiting nonlinearities like those found among frogs, birds and mammals, including primates. By showing that fishes have evolved nonlinear acoustic signalling like all other major lineages of vocal vertebrates, these results suggest strong selection pressure favouring this mechanism to enrich the spectro-temporal content and complexity of vocal signals.     

Regeneration of the caudal and pectoral fins of a bony fish, Gambusia (Haplochilus) schoelleri

The study showed that excision of the caudal fin at basal level was followed by complete regeneration in one and a half months, whereas if it was amputated at mid-fin level, complete regeneration took two months. The findings confirm that the greater the extent of amputation, the faster the rate of regeneration. In the case of the pectoral fin, only part of which was removed, it was found that the fin completely regenerated, with recovery of its original pattern, within two months after amputation.     

Molecular,cellular and histological changes in skin from a larval to an adult phenotype during bony fish metamorphosis

Developmental models for skin exist in terrestrial and amphibious vertebrates but there is a lack of information in aquatic vertebrates. We have analysed skin epidermal development of a bony fish (teleost), the most successful group of extant vertebrates. A specific epidermal type I keratin cDNA (hhKer1), which may be a bony-fish-specific adaptation associated with the divergence of skin development (scale formation) compared with other vertebrates, has been cloned and characterized. The expression of hhKer1 and collagen 1α1 in skin taken together with the presence or absence of keratin bundle-like structures have made it possible to distinguish between larval and adult epidermal cells during skin development. The use of a flatfish with a well-defined larval to juvenile transition as a model of skin development has revealed that epidermal larval basal cells differentiate directly to epidermal adult basal cells at the climax of metamorphosis. Moreover, hhKer1 expression is downregulated at the climax of metamorphosis and is inversely correlated with increasing thyroxin levels. We suggest that, whereas early mechanisms of skin development between aquatic and terrestrial vertebrates are conserved, later mechanisms diverge. This work was carried out within the project “Arrested development: The Molecular and Endocrine Basis of Flatfish Metamorphosis” (Q5RS-2002-01192) with financial support from the Commission of the European Communities. It does not necessarily reflect the Commission’s views and in no way anticipates its future policy in this area. This project was further supported by Pluriannual funding to CCMAR by the Portuguese Science and Technology Council. M.A. Campinho was sponsored by the Portuguese Ministry of Science (grant no. SFRH/BD/6133/2001).     

Refined structure of bony fish muscle myosin filaments from low-angle X-ray diffraction data

Application of X-ray diffraction methods to the elucidation of the arrangement of the myosin heads on myosin filaments in resting muscles is made simpler when the muscles themselves are well ordered in 3D. Bony fish muscle for the vertebrates and insect flight muscle for the invertebrates are the muscles of choice for this analysis. The rich, well-sampled, low-angle X-ray diffraction pattern from bony fish muscle has previously been modelled with an R-factor of 3.4% between observed and calculated transforms on the assumption that the two heads in one myosin molecule have the same shape. However, recent evidence from other kinds of analysis of other muscles has shown that this assumption may not be valid. There is evidence that the motor domain of one head in each pair may interact with the neck region of the second head. This possibility has been tested directly in the present analysis which extends the X-ray modelling of fish muscle myosin filaments by permitting independent shape changes of the two heads in one molecule. The new model, with a computed R-factor of 1.19% against 56 independent reflections, shows that in fish muscle also there is a marked asymmetry in the organisation of each head pair.