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Maccuro, S.

Publications and source records attributed to Maccuro, S..

2 recordsLinked to original sources

Ultrasound-based prediction of spawning time in females of Octopus vulgaris

There has been a great interest in common octopus Octopus vulgaris aquaculture over recent years and commercial production is coming within reach. In addition, O. vulgaris is a short-lived semelparous species with a long spawning period. Therefore, monitoring ovary maturation in cultured octopuses can be useful in the scheduling of hatchery operations and management of culture resources. The present work aimed to assess the ovary maturity status in cultured and wild common octopus by means of a noninvasive technique, ultrasonography, to anticipate the time of spawning in aquaculture. A total of 27 octopuses (15 cultured and 12 wild ones) in different ovarian maturation stages were used in ultrasonographic procedures. Ultrasound-based ovary volumes were compared with real ovarian volumes and then the ultrasonography-based gonadosomatic index (GSIU) was estimated. Data from cultured females indicate that GSIU was initially below 0.3 cm3 per 100 g of body weight in immature animals, and attained a fast increase until reaching about 4.0 cm3 per 100 g of body weight right before spawning. The steep increase in relative volume occurred similarly in females of different body weights. In addition, the specific growth rate of cultured animals decreases with time, and with GSIU. Lastly, a linear mixed model indicated a significantly negative and logarithmic relationship between the time to spawning and GSIU in captive females. That relationship will be useful to predict spawning time in immature and maturing females, helping in the management of octopus broodstocks in research and production facilities.

zoology↗

Optimization of Whole Mount RNA multiplexed in situ Hybridization Chain Reaction with Immunohistochemistry, Clearing and Imaging to visualize octopus neurogenesis

Gene expression analysis has been instrumental to understand the function of key factors during embryonic development of many species. Marker analysis is also used as a tool to investigate organ functioning and disease progression. As these processes happen in three dimensions, the development of technologies that enable detection of gene expression in the whole organ or embryo is essential. Here, we describe an optimized protocol of whole mount multiplexed RNA in situ hybridization chain reaction version 3.0 (HCR v3.0) in combination with immunohistochemistry (IHC), followed by fructose-glycerol clearing and light sheet fluorescence microscopy (LSFM) imaging on whole-mount Octopus vulgaris embryos. We developed a code to automate probe design which can be applied for designing HCR v3.0 type probe pairs for fluorescent in situ mRNA visualization. As proof of concept, neuronal (Ov-elav) and glial (Ov-apolpp) markers were used for multiplexed HCR v3.0. Neural progenitor (Ov-ascl1) and precursor (Ov-neuroD) markers were combined with an immunostaining for phosphorylated-histone H3, a marker for mitosis. After comparing several tissue clearing methods, fructose-glycerol clearing was found optimal in preserving the fluorescent signal of HCR v3.0. The expression that was observed in whole-mount octopus embryos matched with the previous expression data gathered from paraffin-embedded transverse sections. Three-dimensional reconstruction revealed additional spatial organization that had not been discovered using two-dimensional methods.

developmental biology↗