Molecular and Immunologic Characterization of Gynogenetic Channel Catfish (Ictalurus punctatus)

Second-generation gynogenetic channel catfish were characterized by molecular and immunologic assays to determine if they were isogenic at major histocompatibility complex loci. Southern blot analyses, using channel catfish MHC class II B and class I A gene probes, revealed identical banding patterns among second-generation gynogenetic fish. In contrast, banding patterns from outbred fish differed not only from gynogenetic animals, but also among themselves. Nucleotide sequence analysis of the MHC class II β₁ domain, which encompasses the peptide binding region, from four randomly selected gynogenetic fish showed a single DNA sequence. In contrast, analysis of the same region from three outbred fish showed sequences that differed not only among themselves, but also from those of gynogenetic animals. In cytotoxic assays, peripheral blood leukocytes from outbred fish lysed both gynogenetic and allogeneic targets, whereas those from gynogenetic fish lysed only allogeneic targets. Taken together, these results suggest that this group of second-generation gynogenetic channel catfish is isogenic at MHC loci and may provide an excellent system with which to study cell-mediated immunity in teleosts. Received September 11, 1998; accepted January 14, 1999     

Ictalurus melas (Rafinesque, 1820) and I. nebulosus (Lesueur, 1819): the North American catfishes in Europe

The greater part of the literature on European fishes reports the widespread occurrence of an introduced North American catfish and identifies it as Ictalurus nebulosus. Study of the literature reporting critical determinations and of specimens from Europe and Great Britain reveals the presence of two species, I nebulosus and I melas. These fishes are widely used in experimental studies, usually being obtained through aquarium-fish dealers indirectly from continental Europe. Mostly they are incorrectly identified as I nebulosus. There is reason to believe that I melas is the more commonly imported catfish in Britain; both species occur feral in mainland Europe.     

A study of haematological profiles of channel catfish, Ictalurus punctatus (Rafinesque)

A blood profile of Ictalurus punctatus was determined from a total of 35 fish. The values obtained included plasma protein, haemoglobin, red cell counts and differential white cell counts.     

Ultrastructure of the liver in channel catfish Ictalurus punctatus (Rafinesque)

Livers of 38 channel catfish were studied by light and electron microscopy and the morphology of hepatocytes, endothelial cells, Kupffer cells, fat-storing cells, macrophages, exocrine pancreatic cells, and epithelium of bile pre-ductules and ducts was described, Hepatocytes contained large peri-canalicular bodies which showed acid hydrolase activity. A comparison between catfish liver and that of other teleosts was made. The applicability of the findings to pathobiological studies of teleost liver is described.     

Polymorphism of myofibrillar proteins in histochemically identified myotomal muscle fibre types of Heteropneustes fossilis (Bloch) and Labeo rohita (Hamilton)

Myofibrillar proteins in the myotomal muscle fibre types of two freshwater teleosts, Heteropneustes fossilis (Bloch) and Labeo rohita (Hamilton), were investigated. The fibre types were identified histochemically based on the reactivities of the enzymes SDH and m -ATPase. Electrophoretic analysis revealed distribution patterns of myosin light chains, tropomyosin, troponin, c - and m -protein; specific to the muscle fibre types. The results correlate well with the general pattern of myofibrillar protein distribution found in the skeletal muscles of higher vertebrates. The significance of the present findings are discussed with regard to histochemical, biochemical as well as functional properties of the muscle fibre types.     

Erythrocyte nuclear measurements of diploid and triploid channel catfish, Ictalurus punctatus (Rafinesque)

Minor axis, major axis, and volume measurements of erythrocyte nuclei were compared by discriminant analysis to determine which variable best predicted ploidy levels. Predicted correct classification percentages for minor axis, volume, major axis, and all variables combined were 81·33, 86·21, 92·36 and 92·65%, respectively. Minor axis and volume measurements were not as accurate in predicting ploidy levels because nuclear shape changed as the ploidy level changed from the diploid to triploid state. Mean major axis measurements alone classified 92·36% of the fish correctly with a discriminant level of 4·726 μm.     

Novel heme-binding component in the serum of the channel catfish (Ictalurus punctatus)

The serum of the channel catfish (Ictalurus punctatus) was examined for heme- and hemoglobin-binding proteins. Electrophoretic mobility retardation assays failed to detect a hemoglobin-binding material similar to mammalian haptoglobin; however, a heme-binding component (not previously described) was identified in catfish seru. The heme-binding component was purified by gel filtration chromatography; electrophoretic analyses suggested it to be composed of two polypeptide subunits of molecular masses about 115 and 98 kDa. This composition is inconsistent with hemopexin, the known heme-binding serum protein of mammals. Although it was not fully saturated with heme, the catfish component contained detectable heme in normal sera. When complexed by the binding material, heme was used as an iron source by isolates of the bacterial Gram-negative genusAeromonas; the capacity of other bacteria to use the complex was not tested. The physiological function of the catfish heme-binding serum protein is presently not clear.     

Morphometric properties and innervation of muscle compartments in rat medial gastrocnemius

The rat medial gastrocnemius (MG) muscle is composed of the proximal and distal compartments. In this study, morphometric properties of the compartments and their muscle fibres at five levels of the muscle length and the innervation pattern of these compartments from lumbar segments were investigated. The size and number of muscle fibres in the compartments were different. The proximal compartment at the largest cross section (25% of the muscle length) had 34% smaller cross-sectional area but contained a slightly higher number of muscle fibres (max. 5521 vs. 5360) in comparison to data for the distal compartment which had the largest cross-sectional area at 40% of the muscle length. The muscle fibre diameters revealed a clear tendency within both compartments to increase along the muscle (from the knee to the Achilles tendon) up to 46.9?μm in the proximal compartment and 58.4?μm in the distal one. The maximal tetanic and single twitch force evoked by stimulation of L4, L5, and L6 ventral roots in whole muscle and compartments were measured. The MG was innervated from L4 and L5, only L5, or L5 and L6 segments. The proximal compartment was innervated by axons from L5 or L5 and L4, and the distal one from L5, L5 and L6, or L5 and L4 segments. The forces produced by the compartments summed non-linearly. The tetanic forces of the proximal and distal compartments amounted to 2.24 and 4.86?N, respectively, and their algebraic sums were 11% higher than the whole muscle force (6.37?N).     

Comparison between serum values of pond and intensive raceway cultured channel catfish Ictalurus punctatus (Rafinesque)

Blood serum components in channel catfish ( Ictalurus punctatus ) were compared between raceway (with low dissolved oxygen) and pond populations. Normal limits were defined for the pond populations, for each of 18 variables, as ± two standard deviations. Carbon dioxide, blood urea nitrogen, total protein, uric acid, creatinine, sodium, and total bilirubin were all lower in fish raised in raceways. Carbon dioxide and sodium had the greatest reductions; all reductions were statistically significant.
Alkaline phosphatase, inorganic phosphorus, calcium, cholesterol, and glucose values were higher in raceway fish, but only alkaline phosphatase and glucose were significantly higher.     

Use of shape analysis in the investigation of disputable meristic characters for Ameiurus melas (Rafinesque, 1820) and Ameiurus nebulosus (Lesueur, 1819)

The aim of this paper was to use geometric morphometry as a helpful tool to clarify the overlap between meristic characters (such as the number of anal fin rays) of different fish identification keys employed for the two sister species, Ameiurus melas and Ameiurus nebulosus. Geometric morphometry included shape analysis of 15 landmarks on 432 specimens. The analyses showed significant differences among groups based on the sampling site, while there was no difference among groups based on the number of anal fin rays (used to distinguish species). This suggests that the use of meristic characters for the determination of these two sister species is rather questionable.     

Evidence for a morphological component in acid-base regulation during environmental hypercapnia in the brown bullhead (Ictalurus nebulosus)

Summary Exposure of adult brown bullheads Ictalurus nebulosus (120–450 g) to environmental hypercapnia (2% carbon dioxide in air) and subsequent recovery caused transient changes in whole body net sodium flux (J _net ^Na+ ) and net chloride flux (J _net ^Cl- ) resulting largely from changes in whole body sodium influx (J _in ^Na+ ) and chloride influx (J _in ^Cl- ). Scanning electron microscopy (SEM) revealed that the fractional area of chloride cells (CCs) on the interlamellar regions was reduced by 95% during environmental hypercapnia. During post-hypercapnic recovery, gill filament CC fractional area increased. The changes in J _in ^Cl- during and after environmental hypercapnia were closely associated with the changes in CC fractional area while the changes in J _in ^Na+ did not correspond to the changes in CC fractional area. Transmission electron microscopy (TEM) supported the SEM observations of CC surface area changes and demonstrated that these changes were caused by covering/uncovering by adjacent pavement cells (PVCs). Lamellar and filament PVC microvilli density increased during hypercapnia while there was a subsequent reduction in the post-hypercapnic period. These data suggest that an important mechanism of acid-base regulation during hypercapnic acidosis is modification of the chloride cell-associated Cl^-/HCO ₃ ^- exchange mechanism. We suggest that bullheads vary availability, and thus functional activity, of this transporter via reversible morphological alterations of the gill epithelium. The increase in density of PVC microvilli may be associated with sodium uptake and/or acidic equivalent excretion during acidosis.     

Ionic transporting systems of skeletal muscle in relation with innervation and their involvement in myotonic diseases

Excitation-contraction coupling describes the series of events that begins with propagated action potential on the muscle fiber surface membrane and leads to the twitch contraction of the fiber. The generation of an action potential during excitation requires rapid sequential changes in membrane conductances of Na⁺, Ca²⁺, and K⁺ ions that depend upon the opening and closing of the respective channels. Myotonic disorders are inherited diseases whose clinical manifestations include electrophysiological signs such as increased excitability and delayed relaxation of the muscles after voluntary contraction. All these disorders appears to be due to an abnormality of the muscle itself since they persist after section or blocking of the motor nerve after curarization. Most experimental and clinical data suggest that human myotonia arises from genetically-induced structural and functional alterations of the muscle membrane. The purpose of this article is to focus on the more recent developments in the molecular and pharmacological analysis of cation transporting systems such as ionic channels and (Na⁺, K⁺) ATPase in myotonic disorders.Special issue dedicated to Dr. Lawrence Austin.     

The claw closer muscle of Neohelice granulata (Grapsoidea,Varunidae): a morphological and histochemical study

Longo, M.V., Goldemberg, A.L. and Díaz, A.O. 2011. The claw closer muscle of Neohelice granulata (Grapsoidea, Varunidae): a morphological and histochemical study. —Acta Zoologica (Stockholm) 92 : 126–133. The claw closer muscle of Neohelice granulata was studied according to histological, histochemical, and morphometrical criteria. Adult male crabs in intermoult stage were collected from Mar Chiquita Lagoon (Buenos Aires, Argentina). Muscle fibers show evident striations and oval‐elongated nuclei with loose chromatin. The loose connective tissue among muscle fibers consists of cells and fibers embedded in an amorphous substance. Muscle histochemistry reveals two slow fiber types: ‘A’ and ‘B’. Prevailing A fibers are larger, and they usually show, with respect to B type, a weaker reaction to whole techniques. Fibers with short (SS), intermediate (IS), and long sarcomeres (LS) appear in the claw closer muscle, being the LS fibers predominant. Concluding, the histochemical and morphometrical characteristics of the claw closer muscle fibers of N. granulata are indicative of slow fibers. The slow A type (low resistant to fatigue) prevails.     

Comparative study of mannose-binding C-type lectin isolated from channel catfish (Ictalurus punctatus) and blue catfish (Ictalurus furcatus)

Mannose-binding C-type lectin (MBL) was isolated from channel catfish (Ictalurus punctatus) NWAC 102 and 103 strains, blue catfish (Ictalurus furcatus) D+B and Rio Grande strains, hybrid catfish (channel catfish female NWAC 103 x blue catfish male D+B) sera, and purified by affinity chromatography from channel catfish Norris strain serum. Reduction of purified channel catfish MBL with 2-ME yielded a single band of 62 kDa by SDS-PAGE and Western blot analysis using guinea pig anti-MBL IgG as primary antibody. Channel catfish NWAC 102 strain, channel catfish NWAC 103 strain and hybrid catfish sera had molecular masses of 63 kDa for MBL. Blue catfish (D+B strain) serum MBL had a molecular mass of 66 kDa. Rio Grande blue catfish serum MBL had a molecular mass of 65 kDa. Amino acid composition analysis (mol%) of the affinity-purified channel catfish MBL found a high content of serine present. Functional binding studies of channel catfish and blue catfish MBLs binding to Edwardsiella ictaluri were done using a dot-immunoblot ELISA method. A dot-immunoblot ELISA binding assay was done to compare nine different strains and species of channel catfish and blue catfish for their levels of serum MBL. Blue catfish had higher levels of MBL than did the various strains of channel catfish tested. MBL could be used as a genetic marker for selection of disease resistance in the different strains of catfish used in aquaculture. This study describes the presence of serum MBL in catfish and evidence for a C-type lectin complement pathway of innate immunity.     

The innervation of the myometrium of the cynomolgus monkey (Macaca fascicularis)

Summary The autonomic innervation of the myometrium of Macaca fascicularis consists of bundles of unmyelinated nerve fibres running between the smooth muscle cells, and is therefore considered to be of the fascicular (= unitary) type. Close contacts between nerve fibres and smooth muscle fibres were not found. Modification of the chromaffin method according to Tranzer and Richards made it possible to visualize the heterogeneity of the nerve fibres in a single bundle. The following fibre types were found to coexist: (1) noradrenergic fibres containing synaptic vesicles with a dense granule, (2) cholinergic fibres containing empty synaptic vesicles, and (3) non-adrenergic noncholinergic (NANC) fibres containing only or predominantly large dense-cored vesicles, which do not react with this method. Noradrenergic fibres are the most numerous (around 60%), followed by NANC fibres (30%) and cholinergic elements (around 10%). The distribution of these three types is similar in the cervix, the isthmus and the body of the uterus in pregnant and non-pregnant females.