Outbreeding depression in the common frog, <Emphasis Type="Italic">Rana temporaria</Emphasis>

Theory suggests that parental relatedness is a continuous variable with a fitness optimum that we heretoforth will refer to as optimal outbreeding. In the present paper, we test this proposition from a conservation (translocation) perspective. Amphibians are facing a global decline and many amphibian populations are today small and threatened by extinction. Because genetic differentiation is often high between amphibian populations, they could be particularly sensitive to outbreeding depression, e.g. due to breakdown of locally adapted gene complexes. We tested if outbreeding would reduce fitness in common frogs, Rana temporaria, crossed from a large and an isolated, small population, separated by 130km, using artificial fertilization. For females from the large population, tadpoles were significantly smaller and more malformed in crosses with males from the small population, than with males from the large population. For the small population, however, no significant paternal genetic effects could be found. The difference in response to outbreeding between populations was accompanied with significant differences in the importance of maternal effects. We conclude that care should be taken when translocating frogs between distantly related populations to avoid outbreeding depression.     

Spectro-temporal receptive fields of auditory neurons in the grassfrog

The stimulus-event relation of single units in the auditory midbrain area, the torus semicircularis, of the anaesthetized grassfrog (Rana temporaria L.) during stimulation with a wide ensemble of natural stimuli, was analysed using first and second order statistical analysis techniques. The average stimulus preceding the occurrence of action potentials, in general, did not prove to give very informative results. The second order procedure consisted in the determination of the average dynamic power spectrum of the pre-event stimuli, following procedures as described elsewhere (Aertsen and Johannesma, 1980; Aertsen et al., 1980). The outcome of this analysis was filtered with the overall power spectrum of the complete stimulus ensemble in order to correct for its non-uniform spectral composition. The stimulus-filtered average pre-event dynamic spectrum gives a first indication of the spectro-temporal receptive field of a neuron under natural stimulus conditions. Results for a limited number of recordings are presented and, globally, compared to the outcome of an analogous analysis of experiments with tonal stimuli.     

Lipidkugeln im Plexusepithel von Rana temporaria L.

Zusammenfassung Das Plexusepithel von Rana temporaria L. enthält zwei verschiedene Typen von Lipidkugeln, die einen Durchmesser bis zu 8 erreichen können. Diese Einschlüsse haben keine Zytomembran. Die Struktur der Einschlußkörper wird mit Hinweisen auf die Fixierung und den Stoffwechsel diskutiert.

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Lipid droplets in the epithelium of the choroid plexus of Rana temporaria L.

Summary The epithelium of the choroid plexus of Rana temporaria L. contains two different types of spherical lipid inclusions up to 8 in diameter. The droplets do not have a limiting membrane. The preservation and metabolism of the lipid bodies are discussed.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Dihydropteridine reductase variation in man and the characid fish “Cheirodon axelrodi”: Evidence for a dimeric enzyme structure

Summary Genetic evidence for a dimeric structure of dihydropteridine reductase in man and in the fish species Cheirodon axelrodi and Salmo irideus is presented. A single locus in man and two loci in the fishes examined encode this enzyme. Zymograms revealed two alleles for the locus in man and two alleles for each locus in the fish Cheirodon axelrodi. The liver homogenate of a patient with dihydropteridine reductase deficiency showed no detectable activity in the gel, while his parents showed the normal electrophoretic phenotype.     

Ultrastructural radioautographic analysis of neurogenesis in the hypothalamus of the adult frog,Rana temporaria,with special reference to physiological regeneration of the preoptic nucleus

The localization and fine structure of proliferating cells in the hypothalamic preoptic area were studied by light-and electron-microscopic radioautography 1–2 h following single application of ³H-thymidine to adult Rana temporaria taken from their natural habitat in the spring and autumn. ³H-thymidine uptake by proliferating cells was much more pronounced in frogs caught in May/June, i.e., a month after the breeding period (labeled cells represent about 10% of the total ventricular zone cell population), compared to animals caught in mid-September, when it was very low. In both ³H-thymidine treatment groups the vast majority of labeled cells are found exclusively within the preoptic recess ventricular zone. With regard to ultrastructure, it contained proliferating cells of at least 4 types, ranging from immature forms (bipolar stem cells) to more differentiated elements (tanycyte-like ependymoblasts, classical ependymoblasts). All of them showed label over their nuclei indicating that these cells are capable of DNA synthesis and mitosis. The possible role of the preoptic recess ventricular zone as a source of precursor cells for new peptidergic neurosecretory cells, conventional neurons and glial cells in the hypothalamic preoptic area of the adult frog is discussed.     

Derivatives of Benzotetrazine-1,3-dioxide Are New NO-donors, Activators of Soluble Guanylate Cyclase, and Inhibitors of Platelet Aggregation

The ability of 5-nitro-, 7-nitro-, and 5,7-dinitrobenzotetrazine-1,3-dioxides to generate nitric oxide (NO) and activate soluble guanylate cyclase was investigated. All of these compounds were found to be thiol dependent NO-donors and guanylate cyclase activators. The maximal stimulatory effect of 5-nitro-, 7-nitro-, and 5,7-dinitrobenzotetrazine-1,3-dioxides was observed at 10 M concentration and the activity increase was 4.5-, 15.0-, and 8.2-fold in the presence of 20 M dithiothreitol and 11.3-, 31.6-, and 20.5-fold, respectively, in the presence of added glutathione (100 M). The NO-dependent mechanism of benzotetrazine-1,3-dioxide nitroderivative-induced activation of soluble guanylate cyclase (in the presence of 100 M glutathione) was confirmed by the inhibition (by 78%) of 7-nitrobenzotetrazine-1,3-dioxide (10 M)-stimulated guanylate cyclase activity in the presence of the NO-scavenger-2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (Carboxy-PTIO, 50 M) and by the inhibition with 1H-[1,2,4 ]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 0.3 M) of 5-nitro-, 7-nitro-, and 5,7-dinitrobenzotetrazine-1,3-dioxides (10 M)-stimulated guanylate cyclase by 34, 69, and 39%, respectively. All compounds used inhibited ADP-induced aggregation of human platelets with IC ₅₀ of 10.0, 1.3, and 2.0 M for 5-nitro-, 7-nitro-, and 5,7-dinitrobenzotetrazine-1,3-dioxides, respectively. A clearly defined correlation was established between the ability of the compounds to generate NO, activate soluble guanylate cyclase, and inhibit platelet aggregation.     

Reproductive Biology and Protandrous Hermaphroditism in Acanthopagrus latus

Detailed macroscopic and histological studies of the gonads of a full size and age range of Acanthopagrus latus from each season in Shark Bay, Western Australia, demonstrate that this species is a protandrous hermaphrodite in this large subtropical embayment. Although our scheme for the changes that occur in the ovotestes of A. latus during life is not consistent with some of the conclusions drawn for this species elsewhere, it is similar to that of Pollock (1985 J. Fish. Biol. 26: 301–311) for the congeneric Acanthopagrus australis. The ovotestes of males develop from gonads which contain substantial amounts of both testicular and ovarian tissue. The testicular component of the ovotestes of all males regresses markedly after spawning. During the next spawning season, the ovotestes either become gonads in which the testicular zone again predominates and contains spermatids and spermatozoa (functional males), or gonads in which the ovarian zone now predominates and contains mature oocytes (functional females). Once a fish has become a functional female, it remains a female throughout the rest of its life. In Shark Bay, A. latustypically spawns on a limited number of occasions during a short period in late winter and early spring and has determinate fecundity. The mean potential annual fecundity was ca. 2000000. The total length of 245mm, at which, during the spawning period, 50% of A. latus become identifiable as males, is very similar to the current minimum legal length (MLL) of 250mm, which corresponds to an age of ca. 2.5 years less than that at which 50% of males become females. Thus, although the spawning potential ratio suggests that the present fishing pressure is sustainable, the current MLL should be reviewed if recreational fishing pressure continues to increase.     

Involvement of β-Carotene in the Antioxidant Defense of the Bacterial Cell

Effects of recombinant -carotene on the resistance of E. coli culture to menadione and paraquat were studied. The presence of -carotene in E. coli cells prevented, to a considerable extent, an increase in superoxide dismutase activity (induced by redox mediators) without affecting the culture growth. These findings suggest that -carotene is involved in the defense of cells against oxidative stress.     

Social and reproductive behavior of three mouthbrooding cardinalfishes,Apogon doederleini,A. niger and A. notatus

Synopsis Social and reproductive behavior of three paternal mouthbrooding cardinalfishes (Apogonidae) were investigated in the shallow marine waters of Shirahama, Japan. The solitary species Apogon doederleini and A. niger bred in transient pairs, in which a male and female associated for only a few hours of each afternoon on less than 5 successive days. The prespawning behavior was the same as the courtship display on days prior to spawning. After spawning, egg-incubating males were usually left alone. The gregarious species Apogon notatus formed territorial lasting pairs, which resided at given sites from dawn to dusk on each day during a period of a month or more. After spawning, the egg-incubating male either continued to stay with his mate in the territory, or left it to enter into an aggregation. In the latter case, the female continued to reside in the territory, pairing with a new male whom she brought from an aggregation. It is suggested that in paternal apogonids the prolonged pair bond and territoriality should have developed only in gregarious species as secondary adaptation for reproductive success: to avoid conspecific interference during spawning.     

Does an animal peptide:N-glycanase have the dual role as an enzyme and a carbohydrate-binding protein?

Recently, we have reported purification and characterization of a de-N-glycosylating enzyme, peptide:N-glycanase (PNGase) found in C3H mouse fibroblast L-929 cells, and designated L-929 PNGase [Suzuki T, Seko A, Kitajima K, Inoue Y, Inoue S (1994)J Biol Chem 269, 17611–18]. The unique properties of L-929 PNGase are that the enzyme had a high affinity to the substrate glycopeptide (e.g.K _m=114 µm for fetuin derived glycopentapeptide) and that the PNGase-catalysed reaction is strongly inhibited by the released free oligosaccharides but not by the free peptides formed, suggesting that L-929 PNGase is able to bind to a certain type of carbohydrate chain. In this study, we report the new findings of the mannan-binding property of L-929 PNGase; the de-N-glycosylating enzyme activity of L-929 PNGase was inhibited by yeast mannan and triomannose, Man1 3(Man1 6)Man, but not by mannose and -methyl-d-mannoside. Furthermore, L-929 PNGase was revealed to bind to the glycan moiety of yeast mannan by using mannan-conjugated Sepharose 4B gel as a ligand, suggesting that L-929 PNGase could serve not only as an enzyme but also as a carbohydrate recognition proteinin vivo. Such dual properties found for animal-derived L-929 PNGase are unique and are not shared with other previously characterized plant- and bacterial-origin PNGases — PNGase A and PNGase F, respectively.Abbreviations GLC gas liquid chromatography - GlcNAc-Asn 2-acetamido-1--(l-aspartamido)-1,2-dideoxy-d-glucose - BSA bovine serum albumin - DTT dithiothreitol - EDTA ethylenediaminetetraacetic acid - Gal d-galactose - GlcNAc N-acetyl-d-glucosamine - Man d-mannose; triomannose, Man1 3(Man1 6)Man; - MES 2-(N-morphorino)ethanesulfonic acid - NeuAc N-acetyl-neuraminic acid - PNGase peptide:N ⁴-(N-accetyl-glucosaminyl)asparagine amidase (peptide:N-glycanase,EC 3.5.1.52) - PNP p-nitrophenyl     

Ajuga piskoi (Labiatae) — neu für Jugoslawien

Ajuga piskoi Degen & Baldacci, hitherto known only from the type locality in Albania, has been found in Southern Yugoslavia. Two figures are presented. It is a rare and threatened plant species.

     

Lethal genes on the sex chromosomes concealed in a population of the mosquito Aedes aegypti L.

A population has been examined in which an overall parity between the sexes hides considerable between-family variation in sex ratio. A proportion of families show highly distorted sex ratios, with either an excess of females or an excess of males. Distorted sex ratios are invariably associated with mortality in the immature stages at a level appropriate to the action of recessive lethal genes. It has been shown that 26% of M-bearing (Y) chromosomes and at least 24% of m-bearing (X) chromosomes carry a recessive lethal gene.Two such genes have been investigated. l kills males and, in a cross between two heterozygotes, gives rise to a sex ratio close to 2:1 (excess families). k kills females and, in a cross between two heterozygotes, gives rise to a sex ratio close to 1:2 (excess families). Selection for excess or excess did not increase the level of sex ratio distortion.No crossing over occurs between k and the M/m locus whereas l shows 5–10% recombination with M/m. A test for allelism confirmed that l and k are not allelic. The penetrance of k is complete whereas l shows somewhat less than full penetrance. The penetrance of l has been improved by selection.The high frequency of lethals remained in the population during the two year period of study. There was evidence for heterosis preserving this frequency, the heterozygotes living longer and producing more progeny. However lethals were no longer to be found after four further years of laboratory culture.     

The morphology of cells and cell organelles in the anhydrobiotic tardigrade,Macrobiotus hufelandi

Summary Anhydrobiotic and active hydrated specimens of the tardigradeMacrobiotus hufelandi were investigated by electron microscopy, and the cellular fine structure of both stages was compared. Besides conventional preparation methods, totally anhydrous techniques were used to avoid hydration artefacts in the anhydrobiotic specimens. In the latter the cytoplasm was very electron dense and masked many cell constituents, however, membraneous structures, nuclei, mitochondria, microtubules, and myofilaments could be resolved in the micrographs. After substraction of all peculiarities caused by the preparation methods within the micrographs of cells from anhydrobiotic specimens, the comparison revealed that the basic morphology of the cells and their organelles is not changed in the anhydrobiotic specimens. Maintenance of the structural integrity of its cells is a basic ability of this organism to survive desiccation.This report is part of a doctoral dissertation submitted by the author to the Fakultät für Naturwissenschaften of the University of Heidelberg in 1976. The author would like to thank Prof. Dr. E.Schnepf who guided the dissertation. I would like to thank too the Studienstiftung des deutschen Volkes for general support by a Promotionsstipendium.     

Honey bees as carriers of lychee erinose miteEriophyes litchii (Acari: Eriophyiidae)

Up to 23% of honey bees (Apis mellifera Linnaeus) collected from flowering lychee trees, severely infested with the lychee erinose mite (Eriophyes litchii Keifer), were found to be carrying live mites which were picked up as the bees foraged. This method of lychee erinose mite dispersal is discussed in the context of periodic records of spontaneous infestation of lychee flower panicles from previously uninfested orchards.     

Quantitative relationship between active sodium transport,expansion of endoplasmic reticulum and specialized vacuoles (“scalloped sacs”) in the outermost living cell layer of the frog skin epithelium (Rana temporaria)

Summary When an isolated frog skin (Rana temporaria) is exposed to a hydrostatic pressure difference between inside and outside bathing solutions (inside pressure higher than outside) of 20–50 cm of H₂O and if under these conditions the skin is short-circuited electrically, small vacuoles appear light-microscopically in the outermost living cell layer in the epithelium. The number of such vacuoles shows a linear dependency on the rate of active sodium transport as measured by the short-circuit current. Electron-microscopically, the vacuoles are interpreted as previously undescribed organelles, the scalloped sacs which are about 0.5 in diameter, with a wrinkled surface and bounded by a unit membrane. This organelle is in intimate contact with sacs and tubules of smooth endoplasmic reticulum. The observed increase in the number of scalloped sacs usually is accompanied by a significant expansion of the whole system of endoplasmic reticulum. Some of the vacuoles seen light-microscopically must indeed be expanded cisternae of endoplasmic reticulum. The findings are discussed in light of the possibility that the scalloped sacs and the endoplasmic reticulum may be involved in active transport of sodium ions.     

Immunohistochemical localisation of substances reactive to antisera against α-and β-endorphin and met-enkephalin in the brain of Rana temporaria L.

Summary In the brain of Rana temporaria, two distinct systems reactive with - and -endorphin antisera, respectively, and with a met-enkephalin antiserum, have been detected immunohistochemically.Neurons reacting with - and -endorphin antisera are located (1) in the preoptic nucleus, and (2) in the pars ventralis of the tuber cinereum. Immunoreactive nerve fibres of both groups of perikarya end in the infundibular floor near the capillaries and the preoptico-hypophysial tract. Control reactions have shown that the immunoreactivity is suppressed by the corresponding antigens, but also by -LPH. In view of these results the immunoreactive systems examined correspond to an /-endorphin system or a lipotropinergic system.Neurons reacting with the met-enkephalin antiserum are located in the paraventricular organ. Intense immunofluorescence was observed in the infundibular floor. Controls show that the labelling by met-enkephalin antiserum is exclusively suppressed by met-enkephalin.In the pituitary gland, on the other hand, - and -endorphin antisera reveal: 1) the MSH/ACTH-like cells of the pars intermedia and 2) the ACTH-like cells of the pars distalis.Supported by the D.G.R.S.T., Contrat no 77.7.0648     

Flyingfish Spawning (Parexocoetus brachypterus) in the Northeastern Gulf of Mexico

A spawning aggregation of Parexocoetus brachypterus was observed in the northeastern Gulf of Mexico over the outer continental shelf, 100km south of Mobile, Alabama. The event was considered to be a spawning aggregation given the remarkably high abundance of flyingfish observed, together with unusually vigorous flying, jumping, and swimming activity near the surface, and observations of both males and females exuding ripe sex products when landed. The spawn occurred during May 2001 just after moonrise 2 days post full moon. More than 1000000 flyingfish were estimated to have participated in the spawn. Male flyingfish were three times more abundant than females, consistent with observations of 3–4 flyingfish grouped together before and after the spawning aggregation, if several males were simultaneously pursuing a single female. Although egg attachment to flotsam is the only reproductive mode previously described for flyingfishes with demersal eggs, the P. brachypterus spawn occurred in the absence of flotsam. Other possible egg development modes include egg suspension upon current, mid-water suspension above pycnocline or upon topographically induced turbulence, or benthic.     

Competition and the distribution of spring peeper larvae

Studies of tadpole distributions have shown that despite overlapping affinities for semipermanent and permanent ponds, distributions of the spring peeper (Pseudacris crucifer) and the green frog (Rana clamitans) tend to be nonoverlapping. Because spring peepers are believed to be poor competitors, I hypothesized that competition from green frog larvae limits the distribution of spring peeper larvae. I stocked field enclosures with a constant density of spring peeper larvae, and one of four densities of green frog larvae (a target-neighbor design). Increased green frog density had a small effect on metamorphic size and no effects on survivorship, larval period or growth rates of spring peepers. In contrast to these small interspecific effects, green frogs had a large effect on their own performance. Intraspecific competition resulted in a 50% decline in growth rate and an 11% decline in metamorphic size. These results suggest that the species are segregated in resource use, or that compared with green frogs, spring peepers are better able to cope with depressed resource densities. In either case, this field experiment provides no evidence that interspecific competition from green frogs limits distributions of spring peepers. Other factors such as predation and breeding site choice by adults may contribute to the absence of spring peeper larvae from many semipermanent and permanent ponds.     

Innervation und zentralnervöse Verbindungen des Frontalorgans von Rana temporaria und Rana esculenta

Zusammenfassung Mit neurohistologischen Techniken (Nauta-Verfahren) wurde der Anteil des Stirnorgans von Rana temporaria und Rana esculenta an der zentralnervösen Projektion des lichtempfindlichen Pinealkomplexes geprüft. Nach operativer Unterbrechung des Nervus pinealis im dorsalen Lymphsack lassen sich degenerierende Nervenfasern sowohl im Tractus pinealis als auch im stirnorgannahen Stumpf des Nervus pinealis nachweisen. Die ersteren werden als cerebropetale (afferente), die letzteren als zum Stirnorgan ziehende (efferente) Faserelemente gedeutet. Es ist gelungen, die hirnwärts gerichteten Nervenfasern des Stirnorgans bis in die unmittelbare Umgebung des sekretorischen Subcommissuralorgans zu verfolgen; zerfallende Faserfragmente liegen dicht der Basis des Subcommissuralorgans an. Anders als nach Durchtrennung des Tractus pinealis (vgl. Paul, Hartwig und Oksche, 1971) ließen sich nach Unterbrechung des Nervus pinealis keine Degenerationszeichen im mesencephalen Zentralen Grau darstellen.

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Innervation and central nervous connexions of the frontal organ in Rana temporaria and Rana esculenta Fiber degeneration after surgical interruption of the pineal nerve

Summary The contribution of the frontal organ of Rana temporaria and Rana esculenta to the central nervous projections of the light-sensitive pineal complex has been investigated with neurohistological techniques (Nauta-method). After surgical transection of the pineal nerve within the dorsal lymph sac, degenerating nerve fibers have been observed within the pineal tract and also in the proximal stump of the pineal nerve. Those in the pineal tract have been interpreted as cerebropetal (afferent) connexions of the frontal organ, and those in the pineal nerve as fibers directed towards the frontal organ (efferent elements). The cerebropetal fibers of the frontal organ have been traced to the subcommissural region where they degenerate in close juxtaposition with the secretory subcommissural organ. In contrast to the findings obtained after transection of the pineal tract (see Paul, Hartwig and Oksche, 1971), no degenerating fibers have been observed in the mesencephalic central grey after surgical interruption of the pineal nerve.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Beta-adrenergic control of blood oxygen affinity in acutely hypoxia exposed rainbow trout

Summary Acute exposure of rainbow trout to hypoxic water ( =40 mmHg, 15 °C) caused a significant (P<0.01) increase in blood O₂ affinity, from the normoxicP ₅₀ value (at pH_e 7.93) of 23.2±1.1 mmHg to about 19 mmHg, within 5 min. Specimens injected with the -antagonist propranolol showed no change in bloodP ₅₀, despite a more pronounced reduction of arterial during the hypoxic exposure.The change in bloodP ₅₀ coincided with an increase in plasma catecholamines, notably noradrenaline. There was no change in the molar ratios of ATPHb₄ and GTPHb₄. The altered bloodP ₅₀, however, correlated with an alkalinization and an increased sodium concentration of the red cells. This red cell alkalinization can be explained by -adrenergic stimulation of a membrane bound Na⁺/H⁺ antiporter.Propranolol injection into normoxic resting trout caused a significant decrease in and increase in indicating -adrenergic control of gas exchange in the gills.