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.     

Electron microscopic observations of karyogamy in the fish egg

In the initial step of pronuclear association in fertilized fish eggs, the female and male pronuclei (containing large nucleolus-like bodies) were juxtaposed in the center of the blastodisc and formed nucleoplasmic projections along adjacent surfaces. After contact of the pronuclei, small internuclear bridges joining them were formed by fusion at several regions of the nuclear envelope projections. No specific site of fusion or breakdown of nuclear envelopes was recognized in the pronuclei during karyogamy. In the advanced stage, clumps of condensing chromatin appeared in the nucleoplasm of the newly fused pronuclei. The number and diameter of the internuclear bridges increased gradually by progressive fusion in many regions, finally yielding a spherical zygote nucleus. Following complete formation of the zygote nucleus, the pronuclear envelope began to break down concomitantly with shrinkage of the nucleoplasm, which was highly convoluted around the entire nuclear surface. The nucleoplasm containing chromosomes then mingled with the perinuclear cytoplasm.     

Osmoregulatory physiology of pyloric ceca: regulated and adaptive changes in chinook salmon

Functions of the anatomically obvious, yet peculiar, pyloric ceca of the fish gut have been a source of conjecture for over two millennia since Aristotle hypothesized on digestive utilities. Here, we demonstrate regulated and adaptive changes in osmoregulatory physiology of ceca from chinook salmon (Onchorhynchus tshawytscha). Transfer of salmon from freshwater to seawater (both short- and long-term) significantly stimulated both fluid uptake from 5.1 to 8.3-9.3 microl/cm2/hr and also Na+/K+ -ATPase from 6.5 to 8.3-9.6 micromol/ADP/mg protein/hr. Similar changes were induced with implants of cortisol, which resulted in high physiological cortisol levels in plasma. Ceca, which can number about 200 in chinook salmon, were estimated to account for the majority of fluid uptake capacity of the intestine and, after long-term seawater adaptation, the proportion of uptake capacity was sixfold higher. Transport physiology of ceca is thus under environmental and endocrine control indicative of an important role in salt and water homeostasis.     

Enzyme activity and lipids of a preparation from the pyloric ceca of salmon

Proteolytic activity and lipid composition of enzymic preparations of water extract from the pyloric caeca of salmon fishes have been studied. Phospholipids and sterols are tightly bound with proteins. The participation of lipids in the proteolytic activity is discussed.     

Isolation and characteristics of trypsin from pyloric ceca of the starfish Asterina pectinifera

Trypsin was purified from pyloric ceca of the starfish Asterina Pectinifera by ammonium sulfate precipitation, gel filtration, and cation-exchange chromatography. Final enzyme preparation was nearly homogeneous in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its molecular weight was estimated as approximately 28000. Optimum pH and temperature of A. pectinifera trypsin for hydrolysis of N(alpha)-p-Tosyl-L-arginine methyl ester hydrochloride were approximately pH 8.0 and 55 degrees C, respectively. A. pectinifera trypsin was unstable at above 50 degrees C and below pH 5.0, and was not activated by adding Ca(2+). The N-terminal amino acid sequence of A. pectinifera trypsin, IVGGHEF, was found.     

Dietary phosphorus-responsive genes in the intestine, pyloric ceca, and kidney of rainbow trout

Identification of phosphorus (P)-responsive genes is important in diagnosing the adequacy of dietary P intake well before clinical symptoms arise. The mRNA abundance of selected genes was determined in the intestine, pyloric ceca, and kidney of rainbow trout fed low-P (LP) or sufficient-P (SP) diet for 2, 5, and 20 days. The LP-to-SP ratio (LP/SP) of mRNA abundance was used to evaluate the difference in gene expression between LP and SP fish, and to compare the response with bone and serum P, which are conventional indicators of P status. The LP/SP of intestinal, cecal, and renal type II sodium-phosphate cotransporter (NaPi-II) mRNA abundance changed from approximately 1-2 (day 2) to approximately 1.4-4 (day 5) and to approximately 2-10 (day 20). The LP/SP of renal NaPi-II, vitamin D 24-hydroxylase, and vitamin D receptor mRNA abundance correlated inversely with serum P on day 5 but not on day 2 and day 20. In another study, differentially expressed genes between LP and SP fish were examined by subtractive hybridization, confirmed by Northern blot, and evaluated by t-test and correlation with serum and bone P concentrations. About 30 genes were identified as dietary P responsive at day 20, including intestinal meprin and cysteinesulfinic acid decarboxylase, renal S100 calcium-binding protein and mitochondrial P(i) carrier, and cecal apolipoprotein E, somatomedin B-related protein, and NaPi-II. The LP/SP of mRNA abundance of renal mitochondrial P(i) carrier and intestinal cysteinesulfinic acid decarboxylase changed significantly by day 2, and intestinal meprin by day 5. Hence, these genes and NaPi-II are among the earliest steady-response genes capable of predicting P deficiency well before the onset of clinical deficiency.     

Isolation and characteristics of carboxypeptidase B from the pyloric ceca of the starfish Asterias amurensis

Carboxypeptidase B was purified from the pyloric ceca of the starfish Asterias amurensis. The final enzyme preparation was nearly homogeneous in polyacrylamide gel electrophoresis and its molecular weight was estimated as approximately 34,000. The optimum pH and temperature of the enzyme for hydrolysis of benzoyl-glycyl-L-arginine were at approximately pH 7.5 and 55 degrees C, respectively. The enzyme was unstable at above 50 degrees C and at below pH 5.0. The enzyme was activated by Co(2+), but was inhibited by EDTA and Hg(2+). The N-terminal amino acid sequence of A. amurensis carboxypeptidase B was ASFDYNVYHSYQEIMNWITN.     

An isthmo-optic system in a bony fish

This investigation reveals the existence of an isthmo-optic system in a bony fish for the first time. Two cell aggregates of the isthmic region project bilaterally to each eye in Polypterus. The crossed connections are significantly more developed than the uncrossed ones. These findings provide further evidence for the presence of bilaterally projecting isthmo-optic systems in early stages of vertebrate evolution. Furthermore, they suggest that a loss of one or all of these connections during evolutionary or ontogenetic development reflects a parcellation process as proposed by the parcellation theory.     

Complement system of bony and cartilaginous fish

Accumulating evidence indicates that the complement system experienced a discontinuous development at an early stage of vertebrate evolution. Invertebrates such as echinoderms and ascidians, and the most primitive extant vertebrates, the cyclostomes, seem to have a primitive complement system equipped only with the alternative and lectin pathways. In contrast, cartilaginous fish and higher vertebrates seem to have a modern complement system which has two additional pathways, namely the classical and lytic pathways. Recent molecular analyses of the complement system of bony and cartilaginous fish have not only confirmed the above conclusion, but also revealed a unique characteristic of the complement system of fish, where certain key component genes are duplicated. The complement system seems to play a more pivotal role in body defence in fish, whose adaptive immunity is considered to be at a relatively undeveloped state.     

Long range intermolecular forces between macroscopic bodies: macroscopic and microscopic approaches.

Van der Waals energies of interaction are calculated by two methods, the macroscopic method of Lifshitz and the microscopic method of London-Casimir and Polder-Hamaker for the case of two semi-infinite slabs separated by a thin film. When retardation effects may be neglected, the London-Hamaker approach yields values of dispersion interactions which almost coincide with those of the Lifshitz approach, the magnitude of the former values being larger by approximately 10–25%, which is attributed to the effect of the molecular environment in condensed media. At 50–100 Å film thicknesses where retardation effects are small, dispersion terms are generally the major part of van der Waals forces in the Lifshitz formulation. Hence, for 50–100 Å film thicknesses the Hamaker approach, which only includes dispersion interactions is generally adequate. By accounting for retardation effects, which significantly reduce the magnitude of dispersion interactions at several hundred Å, there is a reasonable agreement between the values obtained by the macroscopic and microscopic approaches. When polar substances are present and for film thicknesses of several hundred Å, where dispersion interactions are significantly reduced, the major contribution to van der Waals forces may arise from orientation and induction terms. For such cases the Hamaker approach may lead to critical underestimates of the calculated magnitude of van der Waals forces. An ad hoc way to overcome this difficulty which is applicable to any geometry is proposed. This study presents a simple procedure for the determination of free energies of interaction between macroscopic bodies of various shapes. The procedure, which is applicable when the molecules of bodies and surrounding medium are isotropic, yields results which closely approximate those obtained with the Lifshitz theory.     

多形类杆菌的电镜观察

Three strains of B. thetaiotaomicron have been isolated from caeca of BALB/c-nu/+ mice of SPF. These Bacteroides are obligately anaerobic, Gram-negative, non-spore-forming and non-motil without flagella rods. A characteristic of staining is deep at ends and stainless at the medium of a rod. The isolation and identification of these strains has been reported in 1987. This paper introduced only the results of EM observations. Under the SEM, the unstained area of rods is always showing a concavity, which is just a nucleoid in sections under the TEM. Many lamellar corpuscles have been found in cell plasma. Some of them have been secreted out of the cells. The chemical properties and physiological functions of them are unknown.     

cDNA cloning and sequencing of phospholipase A2 from the pyloric ceca of the starfish Asterina pectinifera

Three cDNA from the pyloric ceca of the starfish Asterina pectinifera, (namely, cDNA 1, 2, and 3), encoding phospholipase A₂ (PLA₂), were isolated and sequenced. These cDNAs were composed of 415 bp with an open reading frame of 414 bp at nucleotide positions 1–414, which encodes 138 amino acids including N-terminal Met derived from the PCR primer. The amino acid sequence deduced from the cDNA 1 was completely consistent with the sequence determined with the starfish PLA₂ protein, while those deduced from cDNA 2 and cDNA 3 differed at one and twelve amino acid residual positions, respectively, from the sequence of the PLA₂ protein, suggesting the presence of multiple forms in the starfish PLA₂. All of the sequences deduced from cDNA 1, 2, and 3 required two amino acid deletions in pancreatic loop region, and sixteen insertions and three deletions in β-wing region when aligned with the sequence of mammalian pancreatic PLA₂. In phylogenetic tree, the starfish PLA₂ should be classified into an independent group, but hardly to the established groups IA and IB. The characteristic structure in the pancreatic loop and β-wing regions may account for the specific properties of the starfish PLA₂, e.g. the higher activity and characteristic substrate specificity compared with commercially available PLA₂ from porcine pancreas.