Development and application of a novel histological multichrome technique for clam histopathology

Costa, Pedro M.; Costa, Maria Helena

doi:10.1016/j.jip.2012.04.013

Cited by 23 publications

(11 citation statements)

References 15 publications

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“…Histological sections were stained with haematoxylin and eosin to allow visualization of anatomical structures 66 . To visualized proteins and polysaccharides content, sections were also stained with a multichrome technic (Alcian Blue, Periodic Acid–Schiff’s, Haematoxylin and Picric Acid) 67 . Sections were dehydrated with serial baths of alcoholic solutions (50–70–90–100%) and mounted in Eukitt medium.…”

Section: Methodsmentioning

confidence: 99%

Proteinaceous secretion of bioadhesive produced during crawling and settlement of Crassostrea gigas larvae

Foulon

Artigaud

Buscaglia

et al. 2018

Sci Rep

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Bioadhesion of marine organisms has been intensively studied over the last decade because of their ability to attach in various wet environmental conditions and the potential this offers for biotechnology applications. Many marine mollusc species are characterized by a two-phase life history: pelagic larvae settle prior to metamorphosis to a benthic stage. The oyster Crassostrea gigas has been extensively studied for its economic and ecological importance. However, the bioadhesive produced by ready to settle larvae of this species has been little studied. The pediveliger stage of oysters is characterized by the genesis of a specific organ essential for adhesion, the foot. Our scanning electron microscopy and histology analysis revealed that in C. gigas the adhesive is produced by several foot glands. This adhesive is composed of numerous fibres of differing structure, suggesting differences in chemical composition and function. Fourier transformed infrared spectroscopy indicated a mainly proteinaceous composition. Proteomic analysis of footprints was able to identify 42 proteins, among which, one uncharacterized protein was selected on the basis of its pediveliger transcriptome specificity and then located by mRNA in situ hybridization, revealing its potential role during substrate exploration before oyster larva settlement.

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Section: Methodsmentioning

confidence: 99%

Proteinaceous secretion of bioadhesive produced during crawling and settlement of Crassostrea gigas larvae

Foulon

Artigaud

Buscaglia

et al. 2018

Sci Rep

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“…Histopathological alterations were determined in whole soft-body of mussels (n = 6), fixated in Davidson’s solution and embedded in Paraplast. Sections (5 µm) were stained with haematoxylin and eosin (H&E) and the tetrachrome procedure described by Costa and Costa 35 . Semi-quantitative histopathological condition indexes were obtained according to the method described by Costa et al .…”

Section: Methodsmentioning

confidence: 99%

Explorations on the ecological role of toxin secretion and delivery in jawless predatory Polychaeta

Cuevas

Martins

Rodrigo

et al. 2018

Sci Rep

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Motivated by biotechnological prospects, there is increasing evidence that we may just be scraping the tip of the iceberg of poisonous marine invertebrates, among which the Polychaeta are promising candidates for bioprospecting. Here we show that an inconspicuous phyllodocid uses toxins in its uncanny feeding strategy. The worm, a jawless active predator characterised by its bright green colour, preys on larger invertebrates (including conspecifics) by extracting tissue portions with its powerful proboscis through suction. The animal is even able to penetrate through the valves and plates of live molluscs and barnacles. Observations in situ and a series of experiments demonstrated that the worm compensates its simple anatomy with secretion of a novel toxin, or mixture of toxins, referred to by us as “phyllotoxins”. These are carried by mucus and delivered via repeated contact with the tip of the proboscis until the prey is relaxed or immobilised (reversibly). Proteolytic action permeabilises material to toxins and softens tissue to enable extraction by suction. The findings show that toxins are a major ecological trait and therefore play a key role in evolutionary success and diversification of Polychaeta, demonstrating also that understanding adaptative features may become the best showcase for novel animal toxins.

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“…Histological and histochemical analyses, both in paraffin and in resin sections, were performed using a tetrachrome (TC) technique based on Alcian Blue (AB) for acidic sugars (pH 0.5-2.5 to indicate sulphated and carboxylated mucins, respectively), Periodic Acid/ Schiff's (PAS) for neutral polysaccharides, Weigert's iron Haematoxylin (WH) for chromatin, and Picric Acid (PA) for muscle and cytoplasm, following Costa & Costa (2012), with modifications (Costa et al 2014;Rodrigo et al 2015). Resin sections required permeabilisation using a saturated solution of potassium hydroxide in ethanol 100% for 5 min followed by 2 min of 2% m/v borax (sodium borate) just before staining.…”

Section: Methodsmentioning

confidence: 99%

A morphoanatomical approach to the adaptive features of the epidermis and proboscis of a marine Polychaeta: Eulalia viridis (Phyllodocida: Phyllodocidae)

et al. 2018

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Eulalia viridis is a marine Polychaeta of the rocky intertidal that, despite its simple anatomy, is an active predator of much larger invertebrates, from which it extracts pieces of soft tissue through suction. This uncanny feeding strategy triggered the pursuit for the morphological mechanisms that enable adaptation to its environment. The evaluation of the worm anatomy and microanatomy, combining electron and optical microscopy, revealed a series of particular adaptations in the epidermis and in the proboscis (the heavily muscled eversible pharynx). Besides its function in feeding, the proboscis is the main sensory organ, being equipped with numerous sensorial papillae holding chemoreceptors. Additionally, the proboscis possesses tentacles that become exposed when the organ is everted. These provide fast release of mucus and toxins, from mucocytes and special serous cells, respectively (the latter involving both merocrine and apocrine processes), whenever contact with a prey occurs. In its turn, the epidermis provides protection by cuticle and mucus secretion and has a sensorial function that may be associated to the worm's uncommon green pigment cells. Eulalia viridis presents a series of elegant adaptive tools to cope with its environment that are evolutionarily designed to counterbalance its relatively simple body plan.

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Development and application of a novel histological multichrome technique for clam histopathology

Cited by 23 publications

References 15 publications

Proteinaceous secretion of bioadhesive produced during crawling and settlement of Crassostrea gigas larvae

Proteinaceous secretion of bioadhesive produced during crawling and settlement of Crassostrea gigas larvae

Explorations on the ecological role of toxin secretion and delivery in jawless predatory Polychaeta

A morphoanatomical approach to the adaptive features of the epidermis and proboscis of a marine Polychaeta: Eulalia viridis (Phyllodocida: Phyllodocidae)

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