Chapitre 4. An intriguing relationship between Mg/Ca and S/Ca skeletal ratios
3. Perspectives
Il apparaît clairement que la détermination des voies d’incorporation du magnésium est primordiale. Contrairement au calcium, l’utilisation d’isotopes radioactifs permettant une visualisation directe des flux au travers l’épiderme n’est que difficilement applicable dans le cas du magnésium: en effet, les seuls isotopes utilisables de cet élément ont des durées de demi-vie particulièrement courtes (le 28Mg n’a qu’une durée de demi-vie de 21 heures, Lusk & Kennedy 1969). Les voies d’incorporation pourraient être déterminées via l’exposition à différents rapports Mg/Ca. Le magnésium pourrait être détecté en microscopie électronique à transmission couplée à une sonde de micro-analyses à rayons X, une technique déjà utilisée chez les coraux (Marshall 2002, Marshall et al 2007). De manière à éviter les artéfacts chimiques, des techniques de cryofixations et de cryosubstitution permettraient d’observer des coupes de spécimens non décalcifiés. Si elle ne pose pas de problème dans le cas de l’éponge, où l’espace de minéralisation est relativement simple à situer, l’application de cette technique aux échinodermes se révèle de prime abord possible mais extrêmement aléatoire. En effet, chez ces organismes, l’espace de calcification est intradermique et de très petite taille (1 à 5 µm, Ameye et al 1998, 1999), et ne peut être localisé avec certitude comme c’est le cas de l’espace extra-pinacodermal des éponges hypercalcifiées
L’application de la spectrométrie de masse à ionisation secondaire (SIMS et nano-SIMS à haute résolution) sur ces coupes, qui présente les mêmes aléas chez les échinodermes que la précédente méthode, permettrait une visualisation directe des ions dans la cellule et le compartiment de minéralisation (cf Marshall et al 2007 chez les coraux). On pourrait ainsi comparer des spécimens soumis à des incubations dans des eaux de mer de différents rapports Mg/Ca ou à des rapports d’isotopes stables du magnésium et du calcium modifiés.
La détermination du rapport Mg/Ca au niveau du site de calcification résoudrait de nombreuses incertitudes. La minéralisation a-t-elle lieu à un rapport Mg/Ca équivalent à celui de l’eau de mer ou diminué par exclusion du magnésium du site de calcification ? Les colorants fluorescents sont couramment utilisés pour étudier le transport et la régulation des éléments dans les systèmes biologiques. Le Furaptra (mag-fura-2) est un colorant fluorescent liant le magnésium qui permet de déterminer la concentration intracellulaire en ce cation, de manière similaire au FURA-2 pour le calcium (Raju et al 1989, Günther 2006). Il émet une fluorescence proportionnelle à la concentration
Discussion générale
intracellulaire. Nous suggérons des expériences de cultures cellulaires de cellules mésenchymateuses primaires1 de larve d’oursins en présence de ces colorants, à différents rapports Mg/Ca. De même, les larves d’oursins, translucides, permettent des mesures in vivo en microscopie confocale. Néanmoins, l’exactitude de cette technique reste limitée par l’existence d’une certaine affinité du Furaptra pour le calcium.
Une approche moléculaire pourrait être intéressante. En effet, la caractérisation génétique et biochimique de plusieurs transporteurs du magnésium a été réalisée dans une grande diversité d’organismes allant des bactéries (Gardner 2003) jusqu’aux mammifères (Quamme 2010). Le génome de l’oursin ayant été décrit (The Sea Urchin Genome Sequencing Consortium 2006), il serait possible, via leurs séquences, de retrouver quels types de transporteurs sont présents et dans quels tissus ils s’expriment.
1 Les cellules mésenchymateuses primaires sont les seules cellules squelettogènes d’échinoderme qui ont été mises en culture avec succès (pour une synthèse, voir Wilt & Benson 2004)
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