Marine incursions,cryptic species and ecological diversification in Amazonia: the biogeographic history of the croaker genus Plagioscion (Sciaenidae)

Aim We propose a phylogenetic hypothesis for the marine‐derived sciaenid genus Plagioscion in the context of geomorphology and adaptation to freshwaters of South America, and assess the extent to which contemporary freshwater hydrochemical gradients influence diversification within a widely distributed Plagioscion species, Plagioscion squamosissimus. Location Amazon Basin and South America. Methods Using nuclear and mitochondrial DNA sequence data, phylogenetic analyses were conducted on the five nominal Plagioscion species, together with representatives from Pachyurus and Pachypops, using character and model‐based methods. Genealogical relationships and population genetic structure of 152 P. squamosissimus specimens sampled from the five major rivers and three hydrochemical settings/‘colours’ (i.e. white, black and clear water) of the Amazon Basin were assessed. Results Phylogenetic analyses support the monophyly of Plagioscion in South America and identify two putative cryptic species of Plagioscion. Divergence estimates suggest that the Plagioscion ancestor invaded South America via a northern route during the late Oligocene to early Miocene. Within P. squamosissimus a strong association of haplotype and water colour was observed, together with significant population structure detected between water colours. Main conclusions Our analyses of Plagioscion are consistent with a biogeographic scenario of early Miocene marine incursions into South America. Based on our phylogenetic results, the fossil record, geomorphological history and distributional data of extant Plagioscion species, we propose that marine incursions into western Venezuela between the late Oligocene and early Miocene were responsible for the adaptation to freshwaters in Plagioscion species. Following the termination of the marine incursions during the late Miocene and the establishment of the modern Amazon River, Plagioscion experienced a rapid diversification. Plagioscion squamosissimus arose during that time. The formation of the Amazon River probably facilitated population and range expansions for this species. Further, the large‐scale hydrochemical gradients within the Amazon Basin appear to be acting as ecological barriers maintaining population discontinuities in P. squamosissimus even in the face of gene flow. Our results highlight the importance of divergent natural selection through time in the generation and maintenance of sciaenid diversity in Amazonia.     

Immunohistology of gastrointestinal carcinoid tumors

DAKO quick staining kits were used to immunostainings for Epithelial Membrane Antigen (EMA) Neuron Specific Enolase (NSE) and S-100 protein (S-100) in 18 carcinoid tumors of the gastrointestinal tract classified according to Soga and Tazawa. EMA was always absent in carcinoid tumors, being at the same time present within glandular epithelium. 88% of cases showed positive immunostaining for NSE. S-100 immunostaining showed immunopositive stellate cells present within the tumor especially within type A carcinoids. In addition in one carcinoid tumor thick, strongly S-100 positive bundles were noticed at the periphery of nests of tumor cells. Combined immunohistochemical and ultrastructural studies are needed to elucidate interrelation of neoplastic and neural elements within carcinoid tumorsa.     

Tricyclic Imipramine Modification of the Circadian Rhythms of Hypothalamic Serotonin, its Precursors and Acid Catabolite in Individually Housed Rats

Circadian rhythms of serotonin (5HT), its precursors tryptophan (TP) and 5-hydroxy-tryptophan (5HTP) and its acid catabolite 5-hydroxy-indoleacetic acid (5HIAA), were determined in the hypothalamus of control rats and rats which had been treated continuously with subcutaneous imipramine (10 mg/kg/day) for 2 weeks.

Rats were individually housed and entrained to LD12:12. Controls showed the 5HT and TP peaks in the light and dark periods respectively, as reported in the literature, but no inverted correlation (antiphase) between SHT and 5HIAA rhythms.

Imipramine significantly modified circadian rhythm characteristics: the 5HT acrophase was advanced, that of TP and 5HIAA was delayed. Imipramine also significantly increased hypothalamic SHT and TP concentrations.     

Chemical modification of essential carboxyl group and histidine residue in the plasma-membrane 5'-nucleotidase

An investigation, using specific chemical reagents, of the amino acids involved in the catalytic activity of the purified 5'-nucleotidase (5'-ribonucleotide phosphohydrolase, EC 3.1.3.5) from bovine liver plasma membranes, was carried out. The enzyme was irreversibly inactivated by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). The inhibition kinetics were of the first-order type and decreased partially in the presence of nucleotides and divalent cations. These results indicate for the first time that a carboxyl group is essential for the catalytic process of 5'-nucleotidase. Moreover, chemical modification by diethylpyrocarbonate also produced inactivation of the enzyme and showed a differential spectrum with a peak at 240 nm characteristic of N-carbethoxyhistidine residues. This inactivation was efficiently released upon decarbethoxylation by hydroxylamine only when the extent of inactivation, due to low concentration of diethylpyrocarbonate, was limited. The time-dependent inactivation followed first-order kinetics and nucleotides afforded significant protection against diethylpyrocarbonate modification. The results indicate the involvement of the histidine residue in catalysis.     

Amino acid sequence of rabbit ventricular myosin light chain-2: Identity with the slow skeletal muscle isoform

Many studies have established a correlation of differences in the activities of various muscle types with differences in the expression of myosin isoforms. In this paper we report the sequence determination of myosin light chain-2 from rabbit slow skeletal (LC2s) and ventricular (LC2v) nmscles. We sequenced tryptic peptides from LC2v which account for all except a few terminal amino acid residues. The major part (87 residues) of the rabbit LC2s sequence, obtained from tryptic and cyanogen bromide (CNBr) peptides, was found to be identical to rabbit LC2v. Our results provide the first sequence information on LC2s from any species, and lend strong support to the hypothesis that LC2s and LC2v are identical. Comparisons of rabbit LC2v and LC2s with rabbit LC2f (from fast skeletal muscle), and also with chicken LC2f and LC2v, show clearly that LC2s and LC2v from mammalian and avian species are more closely related to each other than they are to LC2f isoforms from the same species.     

Die Diffusion des 59Fe-markierten Hämoglobins,ein Artefakt der Glutaraldehyd-Fixierung

Zusammenfassung Ratten wurden 50–80 c ⁵⁹Fe-Citrat in die caudale Vene injiziert. Nach 3–5tägigem Einbau des markierten Eisens in das Hämoglobin wurden Milz-Blöckchen (2 × 2 × 5 mm) 2 Std in GA vorfixiert oder in Hanks-Lösung (= Kontrolle) überführt. Ein Teil der Blöckchen wurde anschließend in OsO₄-Lösung nachfixiert.Die autoradiographischen Ergebnisse zeigen eine Diffusion des Hämoglobins vom unfixierten Zentrum zur Peripherie des Blöckchens.Aktivitätsbestimmungen, die am ganzen Blöckchen, dessen abgetrennten zentralen und peripheren Anteilen, sowie im Überstand vorgenommen wurden, bestätigen diese Diffusion. Während der OsO₄-Nachfixierung erfolgte ein weiterer Verlust des markierten Hämoglobins aus dem zentralen Teil des Blöckchens, nicht aber aus der vorfixierten Peripherie.

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The diffusion of ⁵⁹Fe-labelled hemoglobin, an artefact of the fixation with glutaraldehyde

Summary Injections of 50–80 c ⁵⁹Fe-citrate into the caudale vein of rats were performed. After 3–5 days of ⁵⁹Fe incorporation into the hemoglobin the spleen was taken off, cut in small blocks (2 × 2 × 5 mm) and prefixed for 2 hours in GA or transfered into Hankssolution (= control). Later on some blocks of the spleen were postfixed in OsO₄ solution.A diffusion of the hemoglobin from the unfixed center to the peripheral tissue of the spleen-block is demonstrated by autoradiographic results.After measuring the radioactivity of the total spleen-block of the separated central and peripherical parts as well as their supernatants a diffusion was confirmed. A further loss of the labelled hemoglobin has been observed during the OsO₄ postfixation from the central part of the spleen-block, but not from the prefixed periphery.