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PubMed1996

Nineteenth century research on naturally occurring cell death and related phenomena.

Research on naturally occurring cell death is older than current opinion gives credit. More than 100 nineteenth century publications deal with it, and we review most of these. Soon after the establishment of the cell theory by Schleiden and Schwann, Carl Vogt (1842) reported cell death in the notochord and adjacent cartilage of metamorphic toads. Subsequent landmark discoveries included the massive cell death that occurs in pupating diptera (Weismann 1864), chondrocyte death during endochondral ossification (Stieda 1872), phagocytosis associated with cell death in the muscles of metamorphic toads (Metschnikoff 1883), chromatolytic (apoptotic) cell death in ovarian follicles (Flemming 1885), the reinterpretation of "Sarkoplasten" as "Sarkolyten" in metamorphic amphibia (Mayer 1886), the programmed loss of an entire population of neurons in fish embryos (Beard 1889), the death of scattered myocytes and myofibres in mammalian muscle (Felix 1889), and the death of many motor and sensory neurons in chick embryos (Collin 1906). Other lines of nineteenth century research established concepts important for understanding cell death, notably trophic interactions between neurons and their targets, and intercellular competition.

Animals↗
PubMed1978

Immunofluorescence of somatostatin-producing sites in the hypothalamus of the tadpole, Alytes obstetricans Laur.

Three sites of somatostatin-synthesizing perikarya, or a related antigen, were determined by immunofluorescence in the hypothalamus of the tadpole, Alytes obstetricans (Amphibia, Anura). Two sites of neurosecretory perikarya were localized in the preoptic nuclei of the anterior hypothalamus; the axons extended either to the anterior diencephalon or to the median eminence and the pituitary. The third site was found in the posterior hypothalamus. These neurosecretory cells showed a strong immunofluorescent reaction; their axons all terminated at the level of the median eminence. Somatostatin cells were only found in intact or hypophysectomized tadpoles given somatotropin (STH). The strong reaction observed in hypophysectomized tadpoles was possibly due to the loss of the terminal portion of the neurosecretory pathway (median eminence and pituitary) by which the agent is transported to the site of discharge.

Amphibians↗
PubMed1984

Delta crystallins and their nucleic acids.

delta-Crystallin is a major structural protein of avian and reptilian lenses that is absent from the lenses of fish, amphibia and mammals. It appears to be a tetrameric protein with a native molecular weight near 200 000 (200K) and polypeptide molecular weight near 50K and 48K) (see Note added in proof). The alpha-crystallin polypeptides are extremely similar, associate in various combinations of four and are held together by hydrophobic interactions. Although principally cytoplasmic, delta-crystallin may associate with the cell membrane. delta-Crystallin differs from other lens crystallins in its alpha-helical content, native and subunit molecular weights, antigenicity, low wavelength of maximum fluorescence emission (315 nm) after excitation at 280 nm and amino acid composition (high in leucine; low in aromatic residues en no cysteine). Analyses of peptides, native and subunit molecular weights, and circular dichroism spectra indicate that the primary, secondary, tertiary and subunit structures of delta-crystallin have been generally conserved during evolution. There are at least two tandemly arranged delta-crystallin containing 13-15 introns in the chicken; a similar structure exists for a cloned delta-crystallin gene in the duck. Experiments with chicken show that delta-crystallin synthesis occurs principally in the embryo, especially during lens fiber cell differentiation. delta-Crystallin synthesis also takes place during lens fiber cell differentiation in culture. There is evidence for both transcriptional and post-transcriptional regulation of delta-crystallin synthesis. Current studies on the crystallographic and primary structures of delta-crystallin, on the structure, evolution and expression of the delta-crystallin genes, and on the translation of delta-crystallin mRNAs make this specialized lens protein an active area of investigation.

Amino Acids↗
PubMed1975

Factors determining decussation at the optic chiasma by developing retinotectal fibres in Xenopus.

In Amphibia all primary retinotectal nerve fibres cross at the chiasma. To investigate why this decussation takes place, a series of embryological operations were performed in which one host eye was replaced by an eye from the opposite side of a donor in Xenopus laevis. The visual projections to the optic tecta were mapped electrophysiologically in these animals when adult to reveal to which side of the brain the nerve fibres had become connected. In the majority of cases the contralaterally grafted eyes had developed primary projections to both tecta, in others all the fibres had innervated the contralateral tectum and in 2 animals optic axons were detected only from the ipsilateral side. In 2 other animals optic nerve fibres from both the normal and operated eyes had innervated both tecta. Control experiments showed that section of the optic nerve and stalk without any misalignment of the eye and stalk did not disturb the normal decussation of the fibres. Neither did enucleation in the embryo affect the decussation of the fibres from the remaining eye. It is concluded that no "side specificity" has been shown to exist to determine the side of the brain into which retinotectal fibres develop; rather it seems that complete decussation depends on the developing optic nerve fibres being able to enter a normally aligned optic stalk as they leave the eye.

Animals↗
PubMed1980

Neuroanatomical identification of the frog habenular connections using peroxidase (HRP).

A study of the habenular nuclei connections by means of horseradish peroxidase (HRP) has never been carried out in amphibia. In the present paper we have investigated the afferent projections of the left and right habenular nuclei of the frog Rana esculenta using this technique. Cells, labelled by HRP, were either in a Golgi-like pattern or in a granular pattern. It appears that the habenular nuclei on the two sides of the epithalamus do not show different connections even though they are morphologically asymmetric. In fact, each habenula is connected bilaterally with the septal area and the bed nucleus of the hippocampal commissure, and ipsilaterally with the hypothalamic areas, the entopeduncular nucleus, the periventricular gray of the third ventricle and the interpeduncular nucleus. However, the habenular commissure has typical commissural fibres which apparently do not involve the medial portion of the left habenula. The habenular connections in the frog are generally similar to those reported in the literature for mammals. In addition, our results show the possibility that HRP is transported both retrograde and anterograde.

Animals↗
PubMed1988

Neurophysiological properties of the retinal ganglion cell classes of the Cuban treefrog, Hyla septentrionalis.

The properties of the retinal ganglion cell classes in the cuban treefrog Hyla septentrionalis were studied qualitatively and quantitatively. In the superficial layers of the optic tectum three main classes of afferent optic nerve fibers could be distinguished, class-1*, class-3 and class-4 neurons. Hyla displays a more "classical" organization of the receptive fields in class-1* neurons and a weaker inhibitory surround and lower thresholds with respect to velocity, size and contrast than in Bufo or ranid frogs. The functions for velocity, contrast, size of stimulus, neuronal adaptation and adaptation to background luminance level were evaluated. Experiments with monochromatic light spots are mentioned. The results are compared to those of other amphibia and the diversity of the retinal ganglion cell properties in the different species is stressed as an important factor in the processing of the various ganglion cell types at the tectal level.

Action Potentials↗
PubMed1982

Chromosomal localization of 18S + 28S and 5S Ribosomal RNA genes in evolutionarily diverse anuran amphibians.

The chromosomal locations of the 18S + 28S and 5S ribosomal RNA genes have been analyzed by in situ hybridization in ten anuran species of different taxonomic positions. The chosen species belong to both primitive and evolved families of the present day Anura. Each examined species has 18s + 28S rRNA genes clustered in one locus per haploid chromosome set: this locus is placed either in an intercalary position or proximal to the centromere, or close to the telomere. The 5S rRNA genes are arranged in clusters which vary in number from one to six per haploid set. The 5S rDNA sites are found in intercalary positions, at the telomeres, and at, or close to, the centromeres. Microchromosomes and small chromosomes in primitive karyotypes have been found to carry 5S rDNA sequences. The results are discussed in relation to ideas on the karyological evolution of Amphibia.

Animals↗
PubMed1995

Localization and coexistence of atrial natriuretic peptide (ANP) and neuropeptide Y (NPY) in vertebrate adrenal chromaffin cells immunoreactive to TH, DBH and PNMT.

Antisera specific for mammalian atrial natriuretic peptide (ANP) and neuropeptide Y (NPY) were applied to examine, in immunofluorescence, the occurrence of cells immunoreactive to ANP and NPY in the adrenal organs of mammals, birds, reptiles, amphibians, and bony fish. Catecholamine-containing cells were identified using antisera against tyrosine-hydroxylase, dopamine-beta-hydroxylase, and phenylethanolamine-N-methyl-transferase. In all vertebrates studied, immunoreactivities to ANP and NPY occurred in adrenal chromaffin cells but were absent from the cortex or its homolog, the interrenal. The majority of immunoreactivities to ANP and NPY was confined to the adrenaline cells. In mammals, the number of ANP-immuno-reactive cells (60%-80% of the total cell population) exceeded that of the NPY-immunoreactive cells (35%-45%). In birds, reptiles, and Amphibia, the numbers of ANP-immunoreactive (35%-40%) and NPY-immunoreactive (30%-35%) cells were in a similar range. The bony fish showed a density of both ANP-immunoreactive (80%-90%) and NPY-immunoreactive (35%-40%) cells. In all species studied, immunoreactivities to ANP and NPY partially coexisted. Generally, 30%-55% of the ANP-immunoreactive cells also contained NPY-immunoreactivity. In rat, coexistence amounted to almost 100% and in quail to 95%. Except for the rat, three subpopulations of chromaffin cells seemed to occur: ANP-immunoreactive non-NPY-immunoreactive, ANP-immunoreactive+NPY-immunoreactive, and NPY-immunoreactive non-ANP-immunoreactive cells. Thus, adrenal ANP and NPY share a conservative history and coexist as early as at the level of bony fish. The endocrine actions of ANP and NPY derived from medullary cells on cortical cells as found in mammals might be based on an ancestoral paracrine system. In submammalians, ANP and NPY may not only act as endocrine hormones, but also influence steroid-producing interrenal cells in a paracrine manner, and act as modulators on chromaffin cells.

Adrenal Medulla↗
PubMed1993

Distribution of immunoreactive Tamm-Horsfall protein in various species in the vertebrate classes.

A sheep antibody to human Tamm-Horsfall protein, the major protein in normal urine, was used in an immunohistological study of organs of 48 species of vertebrate animals, representing the classes Mammalia, Aves, Reptilia, Amphibia, Osteichthyes and Chondrichthyes. Immunoreactivity was shown in the thick limb of the loop of Henle in the kidney of mammals, but there was no reactivity with tissues of birds or reptiles. Superficial layers of the skin of several amphibians and fish, superficial layers of the oral mucosa and gills of fish, and the distal tubules of the kidney of some amphibians, reacted with the antibody. Immunoreactivity with mammalian kidney was removed by passage of the antibody down an immunoadsorption column coated with human Tamm-Horsfall protein, and amphibian immunoreactivity was removed by incubation of the antibody with material prepared from frogs in the same way as Tamm-Horsfall protein. These findings suggest that immunoreactive Tamm-Horsfall protein appeared early in vertebrate phylogeny, initially in skin and gills and later in kidney, and that although conserved in evolution, it shows antigenic differences between amphibians and mammals. Its distribution is consistent with the hypothesis that is acts as a waterproofing agent.

Amphibians↗
PubMed1991

Histone genes are located at the sphere loci of Xenopus lampbrush chromosomes.

In the anuran Xenopus, as has been demonstrated previously in several species of urodele Amphibia, histone genes lie at the sphere organizer loci of the lampbrush chromosomes. They were located by in situ hybridization of a 3H-labelled histone H4 anti-sense cRNA probe applied to lampbrush preparations in which transcript RNA had been retained, and likewise to preparations in which transcripts were absent but whose DNA had been denatured prior to hybridization. In Xenopus the histone genes lie in intimate association with the spheres that are attached to the lampbrush chromosomes, but they are absent from spheres that lie free in the germinal vesicle. The Anura separated from the Urodela several hundred million years ago, so the sphere organizer/histone gene association is of great antiquity. This suggests that the association has a functional significance, though it is one that has yet to be discovered.

Animals↗
PubMed1994

Longevity and antioxidant enzymes, non-enzymatic antioxidants and oxidative stress in the vertebrate lung: a comparative study.

It has been proposed that antioxidants can be longevity determinants in animals. However, no comprehensive study has been conducted to try to relate free radicals with maximum life span. This study compares the lung tissue of various vertebrate species--amphibia, mammals and birds--showing very different and well known maximum life spans and life energy potentials. The lung antioxidant enzymes superoxide dismutase, catalase, Se-dependent and non-Se-dependent glutathione peroxidases, and glutathione reductase showed significantly negative correlations with maximum life span. The same was observed for the lung antioxidants, reduced glutathione and ascorbate. It is concluded that a generalized decrease in tissue antioxidant capacity is a characteristic of longevous species. It is suggested that a low rate of free radical recycling (free-radical generation and scavenging) can be an important factor involved in the evolution of high maximum animal longevities. A low free-radical production could be responsible for a low rate of damage at critical sites such as mitochondrial DNA.

Animals↗
PubMed1975

[The eyespots of the oncomiracidium of Euzetrema knoepffleri (Monogenea): ultrastructure and evolution during the life-cycle (author's transl)].

Euzetrema knoepffleri (Monogenea, Monopisthocotylea) is a parasite of the urinary bladder of Euproctus montanus (Amphibia, Urodela). The eyespots of the oncomiracidium of E. knoepffleri are rhabdomeric; each anterior pigmented cell has one rhabdomere, each posterior pigmented cell has two rhabdomeres. This pattern is quite similar to that of the eyespot of another Monopisthocotylea, Entobdella soleae but the lack of cristalline lens in E. knoepffleri appears as a new feature of the Monopisthocotylea. The symmetry of the pigmented structures seems to be effectively connected with the swimming mode of the larva. Moreover, the ultrastructural differences between the two species Euzetrema knoepffleri and Entobdella soleae may be in relation with their different behaviour concerning light. After the fixation of the larva on its host, the comparative study of the evolution of the eyespots, shows the disappearance of the cristalline lens in Entobdella, and the loss of rhabdomeric structures in Euzetrema. These differences seem related with the nature of the microbiotope of the adult: Entobdella soleae is a skin parasite, Euzetrema knoepffleri a reno-vesical one.

Animals↗
PubMed1982

Evolutionary genetics of birds. V. Genetic distances within Mimidae (mimic thrushes) and vireonidae (Vireos).

Genetic distances (D's) between five species within each of the families Mimidae and Vireonidae were estimated from frequencies of protein electromorphs at 23 loci. For three mimid species in the genus Toxostoma, D equals 0.084 (range, 0.069-0.104); and among three mimid genera, D equals 0.223 (0.167-0.278). These distances typify values previously reported in other birds at comparable levels of taxonomic recognition. In sharp contrast, the mean genetic distance among five congeneric species of Vireonidae is far higher, D = 0.360 (0.027-0.578). One possible explanation for these results is that Vireo species are considerably older, on the average, than are species of Toxostoma or than are members of several other avain genera assayed to date. Conventional thought about the origin and relative age of the Vireonidae appears compatible with this explanation. Although genetic distances in the Vireonidae are large by "avian standards," they remain modest or even small in comparison with distances between many nonavian vertebrate congeners. Results for the Mimidae and the Vireonidae are directly contrasted with genetic distances in well-known genera of Amphibia and Reptilia.

Animals↗
PubMed1976

[Histochemical and biochemical investigation of alpha-glucosidases by means of 2-naphthyl-alpha-D-glucoside (author's transl)].

Histochemical and biochemical studies yield the following method of choice for the in situ detection of neutral (microvillous) and acid (lysosomal) alpha-glucosidases: 12 mg 2-naphthyl-alpha-D-glucoside (dissolved in 0.5 ml N,N-dimethylformamide) and 0.6--0.8 ml hexazonium-p-rosaniline in 10 ml 0.1 M citric acid phosphate buffer for aqueous or 5 ml buffer mixed with equal parts of 2% agar for incubation with semipermeable membranes, pH 5 or 6.5. With this method neutral alpha-glucosidases can be exactly demonstrated in the brush border of the small intestine (glycoamylase, sucrase-isomaltase) and kidney of mammals, birds,fishes, amphibia and reptiles; localization of acid alpha-glucosidases is achieved at the cellular level in many organs and tissues. Fluorometric and photometric measurements prove that 2-naphthyl-alpha-D-glucoside is superior to 6-brom-2-naphthyl-alpha-D-glucoside for the demonstration of alpha-glucosidases in situ due to the lower Michaelis constant and higher maximal reaction velocity of the naphthol derivative.--Among the coupling reagents tested neutral alpha-glucosidases can be localized correctly with hexazotized p-rosaniline (with and without semipermeable membranes) for simultaneous coupling. Fast Blue B delivers false positive results in the suczedaneous and simultaneous coupling procedure using aqueous incubation media; in combination with the membrane technique azo dye can not be observed in the sections. Hexazonium-p-rosaniline inhibits neutral and acid alpha-glucosidases to nearly the same extent as Fast Blue B. Fixation of blocks of tissue in formaldehyde and glutaraldehyde suppresses alpha-glucosidases in the intestine and epididymis. The inhibition rates amount to 50 and 70% respectively. Washing in sugar solution rises enzyme activity to 65 and 50%. Species and organ dependent activity differences of neutral and acid alpha-glucosidases and changes of enzyme activity in the intestine and kidney after castration as well as in the course of pregnancy can be detected by means of biochemistry but not with the histochemical assay including minimal incubation. In comparison with p-nitrophenyl-alpha-D-glucoside the 2-naphthyl derivative is also the substrate of choice for the biochemical determination of alpha-glucosidases.--Agar gel electrophoresis reveals one band in the neutral and acid pH range.

Animals↗
PubMed1985

Ultrastructural distribution of peroxidase in thrombocytes of mammals and submammals.

The localization and distribution of peroxidase (PPO) activity were studied ultracytochemically in thrombocytes from lampreys, carps, frogs, snakes, tortoises, rabbits, sheep, dogs, and monkeys. PPO activity was not detectable in the thrombocytes of lampreys, carps, frogs, and snakes. However, this enzyme activity was demonstrated in the nuclear envelope and endoplasmic reticulum of tortoise thrombocytes. Dog and monkey thrombocytes (blood platelets) exhibited PPO activity in the dense tubular system, but this enzyme activity was not detectable in rabbit and sheep thrombocytes. Our observations are interpreted to suggest that thrombocytes from animals lower than amphibia are peroxidase negative. Furthermore, it can be said that thrombocytes from animals higher than reptiles are generally positive, although there are exceptions. PPO activity was localized in the endoplasmic-reticulum system, but not in the cytoplasmic granules of thrombocytes common to submammals and mammals. In this study, we also compared the distribution of peroxidase activity in thrombocytes, neutrophils, and eosinophils and conclude that these are significant differences in the distribution of PPO and myeloperoxidase.

Animals↗
PubMed1982

Potassium permeability in thin amphibian myelinated fibres.

Potential clamp analysis of thin (10-12 micrometers) myelinated fibres from amphibia (Xenopus laevis and Rana temporaria) showed that they had a large delayed K-permeability (PK) and a PK/PNa-ratio of 0.32, which is similar to the ratio in large (25-30 micrometers) frog fibres. These results agree with earlier evidence of a large delayed PK in 10-15 micrometers Rana fibres; a recent description of a size-dependent difference in PK was thus not confirmed. The large PK (0.32 cm3s-1 X 10(-9) in 10-12 micrometers amphibian fibres contrasted with the small PK (0.05 cm3 s-1 X 10(-9) in 12 micrometers rat fibres. Frog and toad fibres also had a larger nodal capacitance (4 pF), but smaller leak conductance (18 nS) than rat fibres (1.4 pF and 29 nS). These differences may be related to the morphological evidence of a more open node with a larger surface area in the amphibian as compared to the rat node.

Animals↗
PubMed1987

Identification of the conserved, conformation-dependent cytokeratin epitope recognized by monoclonal antibody (lu-5).

The epitope recognized by the murine monoclonal antibody (mAB lu-5) recently described as a formaldehyde-resistant, "pan-epithelial marker" of great value in tumour diagnosis is located on the surface of cytokeratin filaments. It has been preserved during vertebrate evolution from amphibia to man. As this epitope is not reactive after SDS-polyacrylamide gel electrophoresis (SDS-PAGE), the epitope-bearing protein has been identified by a dot-blot antibody binding assay, using purified proteins in which the epitope is reconstituted. We show that the epitope is present in most cytokeratin polypeptides of both the acidic (type I) and basic (type II) subfamily but does not occur in other cytoskeletal proteins. The location of this widespread epitope is discussed with respect to homologies of amino acid sequences of cytokeratins and their conformations.

Antibodies, Monoclonal↗