Elektronenmikroskopische und histochemische Untersuchungen zur Funktion des Kiemenherzanhanges (Pericardialdrüse) von Sepia officinalis

Zusammenfassung Die elektronenmikroskopischen Untersuchungen am Kiemenherzanhang von Sepia officinalis zeigen, daß diesem Organ wie den Venenanhängen exkretorische Funktionen zukommen. Die Pericardialdrüse erweist sich als ein Derivat des Kiemenherzens. Ihre Hauptmasse besteht aus einem polar organisierten Faltenepithel, dessen Ultrastrukturmerkmale (basaler Faltenapparat, Gehalt an Mitochondrien und Lysosomen, sekretorisch aktiver Mikrovillisaum) anderen Exkretionsorganen entsprechend auf aktive Ionentransportvorgänge hindeuten. Es ist dem äußeren Kiemenherzepithel homolog und kommt in ähnlicher Ausbildung auch in anderen Organen (Zentralherz, Kieme) vor. Die polygonalen Zellen der Randzone sind dem zentralen Gewebe des Kiemenherzens zu homologisieren; ihr Reusensystem in Nachbarschaft der Blutlakunen ist ähnlich dem der Podocyten in der Vertebratenniere und macht eine Druckfiltration wahrscheinlich. Als Filtermembranen funktionieren neben dem Plasmalemm auch noch die Basalmembran und Zwischenmembran, die porenartige, für Myofer durchlässige Lücken von 30 Å Weite zeigen. Daneben werden an der Grenze zwischen Epithel und Lakune wie auch in der Intima größerer Gefäße besondere Dichtezellen dargestellt, die ähnlich wie Macrophagen größere Partikel (z.B. Ferritin und Myofer) aufnehmen und intraplasmatisch als dense bodies ablagern.Die enzymhistochemischen Befunde über die Verteilung von G-6-PD, SDH, IDH, MDH und MAO und der sauren und alkalischen Phosphatase in Faltenepithel und Lakunen stützen diese Befunde und sprechen ebenfalls für eine exkretorische Funktion.

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Electron microscopic and histochemical studies on the function of the branchial heart appendage (pericardial gland) in Sepia officinalis (l.)

Summary Our electron microscopic studies of the appendages of the branchial heart and veins of Sepia officinalis suggest an excretory function for these organs. The pericardial gland appears to be a derivate of the branchial heart; it mostly consists of a folded polarized epithelium. Ultrastructural features of the epithelium such as the basal folding system, contents of mitochondria and lysosomes, and secretory microvilli suggest active ion transport in these cells of the pericardial gland. The pericardial gland epithelium is homologous with the external epithelium of the branchial heart; it is also present in a similar form in the central heart and branchia. Polygonal cells present in the peripheral zone of the pericardial gland are homologous with the central branchial heart tissue. The system of basal cell processes of the polygonal cells near blood lacunae is similar to the podocyte-capillary relationship in the vertebrate kidney and suggests the possibility of pressure filtration. The plasmalemma, as well as the basal and intermediate membranes, appear to be filtration membranes; they contain pore-like structures about 30 Å in diameter which are permeable to Myofer. Furthermore, special dense cells are present at the epithelium-lacunae border and in the intima of larger vessels; these cells, similar to macrophages, take up large particles, such as ferritin and Myofer, and deposit them in the cytoplasm as dense bodies.Histochemical findings on the distribution of enzymes G-6-PD, SDH, IDH, MDH, and MAO, and the distribution in the folded epithelium and lacunae of alkaline and acid phosphatases confirm the ultrastructural results and suggest an excretory function of the pericardial gland.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft. — Frau Beyerle-von Wehren sei für wertvolle technische Unterstützung gedankt.     

A triterpenoid and its saponin from Phytolacca esculenta.

A new triterpenoid, esculentagenin, and its glycoside, esculentoside M, were isolated from the roots of Phytolacca esculenta and characterized as 11-oxo-3-O-methyloleanata-12-en-2β,3β,23-trihydroxy-28-oic acid and 3-O-[β - -glucopyranosyl (1→4)-β- -Xylopyranosyl]-28-O-β- -glucopyranosyl-11-oxo-30-methyloleanate-12-en-2β,3β,23-trihydroxy-28-oic acid by spectral and chemical evidence.     

Autophagy-related genes serve as heat shock protein 90 co-chaperones in disease resistance against cassava bacterial blight

Heat shock protein 90 (HSP90) is involved in plant growth and various stress responses via regulating protein homeostasis. Autophagy keeps cellular homeostasis by recycling the components of cellular cytoplasmic constituents. Although they have similar effects on cellular protein homeostasis, the direct association between HSP90 and autophagy signaling remains unclear in plants, especially in tropical crops. In this study, the correlation between HSP90 and autophagy signaling was systematically analyzed by protein–protein interaction in cassava, one of the most important economy fruit in tropic. In addition, their effects on plant disease response and underlying mechanisms in cassava were investigated by functional genomics and genetic phenotype assay. The potential MeHSP90.9-MeSGT1-MeRAR1 chaperone complex interacts with MeATGs and subsequently triggers autophagy signaling, conferring improved disease resistance to cassava bacterial blight (CBB). On the contrary, HSP90 inhibitor and autophagy inhibitor decreased disease resistance against CBB in cassava, and autophagy may be involved in the potential MeHSP90.9-MeSGT1-MeRAR1 chaperone complex-mediated multiple immune responses. This study highlights the precise modulation of autophagy signaling by potential MeHSP90.9-MeSGT1-MeRAR1 chaperone complex in autophagy-mediated disease resistance to CBB.     

Melanin Standard Method: Titrimetric Analysis

Melanin isolated from the ink sac of Sepia officinalis (Sepia melanin) has been proposed as a standard for natural eumelanin, and a standard mild isolation and purification protocol for Sepia melanin has been developed (Zeise, doctoral dissertation, Johns Hopkins University, 1991). The goal of the present work, developed using Sepia melanin, was to quantify the bioavailable carboxylic acid groups present in melanin particles. Bioavailability is governed by the accessibility of carboxy groups to the surrounding biological milieu, and is expressed as microequivalents of carboxy group per gram of melanin. The present work was carried out using an heterogeneous slurry of melanin in a nonaqueous system. A standard acidic titrant, and an automatic titrator operating in an equilibrium titration mode were used to characterize and quantify the carboxy group content of Sepia melanins and several other commonly used melanins purified by a standard method (Zeise et al., Pigment Cell Res. [Suppl] 2:48–53, 1992).     

Functional response of early stages of the cuttlefish Sepia officinalis preying on the mysid Mesopodopsis slabberi

The functional response of Sepia officinalis early stages, preying on mysids of the species Mesopodopsis slabberi was investigated. The effects of five prey densities (12.5, 25, 37.5, 50 and 125 mysids l^-1) and two hatchling ages (1-day-old and 7-day-old) on consumption rate and the frequency of non-feeding animals were tested. Older animals were approximately 50% heavier than newly hatched ones. Hatchlings were individually assayed under 0.25 W m^-2 natural light, 37.8 psu in salinity and 19°C. The effect of prey density on consumption rate was highly significant and no effect of age was detected within the age range tested. Maximal values recorded for consumption rate were about 0.45 mysids h^-1. The frequency of non-feeding individuals was strongly reduced at saturating prey densities. The functional response curve showed an interval of prey densities for which density-dependent prey mortality is probable.     

Molecular analysis of a novel FMRFamide-related peptide gene (SOFaRP(2)) and its expression pattern in the brain of the European cuttlefish Sepia officinalis

FMRFamide-related peptides (FaRPs) are among several neurotransmitters known to regulate the chromatophore function in the European cuttlefish Sepia officinalis. Here we report the cloning and sequencing of a novel S. officinalis FaRP gene (SOFaRP(2)). The complete 835-base pair cDNA sequence of the SOFaRP(2) gene contains an open reading frame of 567 base pairs encoding 188 amino acids and four putative FaRPs, NSLFRFamide, GNLFRFamide, TIFRFamide and PHTPFRFamide. All except TIFRFamide cause chromatophore expansion when assayed in an in vitro chromatophore bioassay. To investigate the expression pattern of SOFaRP(2) gene in the cuttlefish brain, in situ hybridization was performed using a full length RNA probe. The SOFaRP(2) gene was expressed primarily in the posterior chromatophore, anterior chromatophore, lateral basal and optic lobes among other brain locations. The SOFaRP(2) gene appears to be expressed in all brain regions involved in chromatophore regulation. These data suggests that some or all of the four FaRPs encoded by SOFaRP(2) might be involved in controlling chromatophore activity in cuttlefish.     

拟目乌贼幼体日摄食量及其对体重影响

在人工养殖条件下,对拟目乌贼幼体的日摄食量和日增重率进行了分析。结果显示,在水温、盐度、pH等条件相对稳定的环境下,10～15d的拟目乌贼幼体平均日摄食虾苗(体长0.5mm)的条数分别是8、11、12、13、78、83条,在这种摄食量下,拟目乌贼幼体的日增重率分别为6.8%、5.68%、6.44%、8.81%、12.5%。其胴长、体重的生长符合S型曲线生长规律。     

FROM CLONAL TO SEXUAL HYBRIDS: GENETIC RECOMBINATION VIA TRIPLOIDS IN ALL-HYBRID POPULATIONS OF WATER FROGS

Speciation via interspecific hybrids is very rare in animals, as compared to plants. Whereas most plants overcome the problem of meiosis between different chromosome sets by tetraploidization, animal hybrids often escape hybrid sterility by clonal reproduction. This comes at the expense of genetic diversity and the ability to purge deleterious mutations. However, here we show that all-hybrid populations of diploid (LR) and triploid (LLR and LRR) water frogs ( Pelophylax esculentus ) have secondarily acquired sexual reproduction. First, in a crossing experiment analyzed with microsatellite markers, triploid hybrids of both sexes and genotypes (LLR and LRR) recombined their homospecific genomes. Second, the great majority of natural populations investigated had low multilocus linkage disequilibrium, indicating a high recombination rate. As predicted from mating system models, the L genome had constant, low levels of linkage disequilibrium, whereas linkage disequilibrium in the R genome showed a significant reduction with increasing proportion of recombining triploids. This direct evidence of sexual reproduction in P. esculentus calls for a change of the conventional view of hybridogens as clonally reproducing diploids. Rather, hybridogens can be independent sexually reproducing units with an evolutionary potential.     

Antifungal and marker effects of Talisia esculenta lectin on Microsporum canis in vitro

Aims: The purpose of this study was to demonstrate the usefulness of lectin obtained from Talisia esculenta (TEL) seeds as a tool to recognize and study Microsporum canis. For this purpose, we investigated the antifungal and marker action of this lectin and the relationship of these effects with the presence of carbohydrates on the structure of this fungus. Methods and Results: The in vitro antifungal activity of TEL was analysed by broth microdilution assay. In addition, TEL was assessed against the arthroconidia present on hairs obtained from infected dogs and cats. The affinity of fluorescein isothiocyanate (FITC)‐labelled TEL for macroconidia and arthroconidia of M. canis was also tested. The effects of TEL on the growth of the M. canis strains began with 0·125 mg ml^?1, and 100% inhibition was obtained with a concentration of 2 mg ml^?1. The addition of carbohydrates, especially N‐acetyl‐glucosamine and d ‐mannose, inhibited these antifungal effects. TEL was able to inhibit the growth of arthroconidial chitin‐rich forms of M. canis obtained from hairs of infected animals and strains cultured in Sabouraud agar. FITC‐labelled TEL efficiently marked macroconidial and arthroconidial forms of M. canis, as shown by fluorescent microscopy. Conclusions: These results show that the inhibitory effects of TEL on M. canis growth may be related to the interaction of lectin with the carbohydrates present at the micro‐organism’s surface, mainly d ‐mannose and N‐acetyl‐glucosamine. Significance and Impact of the Study: Talisia esculenta can be used as an important tool in the biochemical study of M. canis or as a molecule to recognize this dermatophyte in infected tissue.     

菰黑粉菌的分离鉴定及其发酵液中植物激素的检测

在显微镜下观察孢子形态,并观察单菌落的形态和菌株的微观形态,PCR扩增其ITS-5.8S rDNA序列,测序并在NCBI中进行同源比较,确定其种属。液体培养该菌株,通过高效液相色谱法检测发酵液中植物激素。结果表明：用菌落形态与孢子形态鉴定和分子生物学鉴定的方法,对茭白中分离的一个菌株鉴定为菰黑粉菌,且在其发酵液中检测到植物激素IAA、ABA和GA3,其中IAA含量为0.1306mg/L,ABA含量为0.01367mg/L。