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IMPROVED ISOLATION OF THE CRUSTACEAN HYPERGLYCEMIC HORMONE-CONTAINING GRANULES FROM SINUS GLANDS OF CARCINUS MAENAS (LINNAEUS, 1758)

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An improved neurosecretory granule (NSG) preparation of the shore crab, Carcinus maenas sinus glands, enriched for members of the crustacean hyperglycemic hormone family (CHH) and almost devoid of contaminating subcellular fractions, was obtained by differential and density-gradient centrifugations. CHH was determined in an aliquot of each fraction by means of a specific radioimmunoassay, and purity of the fractions by means of electron microscopy of pellet sections from duplicate tubes. Fraction F3 from the first discontinuous gradient centrifugation was enriched for CHH-containing NSG and purified on a second, shallower discontinuous gradient, and fraction F3b proved to contain solely CHH granules as revealed by EM and RIA criteria.

Eine verbesserte Präparation neurosekretorischer Granula (NSG) aus Sinusdrüsen der Strandkrabbe Carcinus maenas durch Differential- und Dichtegradientenzentrifugation wird dargestellt. Damit wurde eine Anreicherung von Neuropeptiden der Crustaceen hyperglykämischen Hormonfamilie (CHH) erreicht, die weitgehend frei von Kontaminationen anderer subzellulärer Fraktionen ist. CHH wurde jeweils in einem Aliquot der Fraktionen durch ein spezifisches Radioimmunoassay bestimmt. Die Reinheit der Fraktionen wurde elektronenmikroskopisch in Pellets nachgewiesen, die aus parallelen Ansätzen gewonnen wurden. Die Fraktion F3 der ersten diskontinuierlichen Gradientenzentrifugation ergab angereicherte CHH-enthaltende NSG und wurde über einen zweiten flacheren diskontinuierlichen Gradienten gereinigt. Die Fraktion F3b enthielt ausschließlich CHH-Granula nach den Kriterien der EM und RIA Untersuchungen.

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/content/journals/10.1163/156854000504165
2000-02-01
2016-09-25

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