a-whole-mount-situ-hybridization-method-for-gastropod-mollusc-lymnaea

A Whole Mount <em>In Situ</em> Hybridization Method for the Gastropod Mollusc <em>Lymnaea stagnalis</em>

Whole mount in situ hybridization (WMISH) is a technique that allows for the spatial resolution of nucleic acid molecules (often mRNAs) within a ...

Georg-August University of Göttingen

The morphological variety displayed by the molluscan shell underlies much of the evolutionary success of this phylum. However, the broad diversity of shell forms, sizes, ornamentations and functions contrasts with a deep conservation of early cell movements associated with the initiation of shell construction. This process begins during early embryogenesis with a thickening of an ectodermal, 'dorsal' (opposite the blastopore) population of cells, which then invaginates into the blastocoel to form the shell gland. The shell gland evaginates to form the shell field, which then expands and further differentiates to eventually become the adult shell-secreting organ commonly known as the mantle. Despite the deep conservation of the early shell forming developmental program across molluscan classes, little is known about the fine-scale cellular or molecular processes that underlie molluscan shell development.

An ancient process in a modern mollusc: early development of the shell in Lymnaea stagnalis.

The ability to visualise the expression of individual genes in situ is an invaluable tool for developmental and evolutionary biologists; it allows for the characterisation of gene function, gene regulation and through inter-specific comparisons, the evolutionary history of unique morphological features. For well-established model organisms (e.g., flies, worms, sea urchins) this technique has been optimised to an extent where it can be automated for high-throughput analyses. While the overall concept of in situ hybridisation is simple (hybridise a single-stranded, labelled nucleic acid probe complementary to a target of interest, and then detect the label immunologically using colorimetric or fluorescent methods), there are many parameters in the technique that can significantly affect the final result. Furthermore, due to variation in the biochemical and biophysical properties of different cells and tissues, an in situ technique optimised for one species is often not suitable for another, and often varies depending on the ontogenetic stage within a species.

Article	Year
An ancient process in a modern mollusc: early development of the shell in Lymnaea stagnalis. BMC developmental biology\| PubMed ID: 23845038	2013
An optimised whole mount in situ hybridisation protocol for the mollusc Lymnaea stagnalis. BMC developmental biology\| PubMed ID: 25879449	2015

Jennifer Hohagen

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