ARE SQUID STATOLITHS HOLLOW DURING EMBRYOGENESIS?
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Squid statoliths are
paired calcareous concretions essentially composed of calcium carbonate
crystallised as aragoniteand found inside cranial fluid-filled cavities, the
statocysts – the main organ of detection of gravity and movement. Statolith use
as an ageing tool was validated in the 1980s, when the daily periodicity of the
growth increments was confirmed by chemically marking these structures in
animals maintained in aquaria. Although statolith analysis has brought major
advances in the knowledge of squid population-dynamics, little is known about
the processes involved in statolith’s calcification and increment deposition
(e.g. during embryogenesis). According to Villanueva (2000), embryonic
statolith development involves the formation of structures such as the focus,
nucleus, postnuclear zone and natal ring. The aim of this study was to
investigatethe microstructure of the embryonic statolith, considering all main
structures,in recently-hatched squids using optical (fluorescence) microscopy,
scanning electron microscopy (SEM) and confocal microscopy. Recently-spawned
egg masses of the European squid, Loligo vulgaris, were collected in the west
coast of Portugal. After hatching at 13-15ºC, statoliths were removed from the
paralarvae andpreserved at -20ºC. Prior to confocal analysis, statoliths were
submitted to Alizarin red S (ARS) staining (5% of ARS in 0.1% KOH) to verify
the presence of calcium content. In addition, images from optical microscopy
and scanning electron microscopy (SEM) were acquired. The findings obtained by
auto-fluorescence observation (Figure 1 A, B) and SEM (Figure 1 C), show that
the embryonic statolith is hollow in some of the area between the nucleus and
the natal ring. ARS stained samples analysed in confocal microscopy (Figure 2
A-G), confirmed the previous observation. This surprising structural feature in
the origin of CaCO3 sensorial structures has never been described before and
suggests a production, with minimum of material and energy expenditure. This is
fundamentally interesting not only in biological terms but also considering
biotemplating orbiomimetics approaches (i.e. synthetic approaches), where
hollow calcium carbonate structures have attracted considerable attention owing
to their unique structural, optical, and surface properties that lead them to a
wide range of applications, including as templates for functional architecture
ARE SQUID STATOLITHS HOLLOW DURING EMBRYOGENESIS?
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