Effects of wall vessel rotation on the growth of larval zebrafish inner ear otoliths

Abstract

Stimulus dependence is a general feature of developing sensory sytems. It has been shown earlier that the growth of otoliths of late-stage Cichlid fish (Oreochromis mossambicus) and Zebrafish (Danio rerio) was slowed down by hypergravity, whereas microgravity during spaceflight yielded an opposite effect, i.e., larger than 1 g otoliths, in Swordtail (Xiphophorus helleri) late-stage embryos. Using ground-based techniques to apply simulated weightlessness, long-term clinorotation (exposure on a fast-rotating clinostat with one axis of rotation for 7 days) led to larger than 1 g otoliths in late-stage Cichlid fish, which is fully in line with the results obtained on Swordtails from spaceflight. Hitherto, early-staged fish have not yet been subjected to (simulated or real) long-term (i.e., more than 3 or 4 days) weightlessness to investigate otolith growth. The present study was carried out in order to fill this gap. Therefore, we subjected Zebrafish at a somite-stage to Wall Vessel Rotation (WVR; a method regarded to provide simulated weightlessness), when the anlage of the inner ear already is present (10 h post fertilisation, hpf). Siblings were maintained under WVR for 3, 6, 9 and 12 days. Further short-term experiments (3 days) were carried out on 10 hpf animals as well as on very early larvae (1 K cell stage, 3 hpf) at two different rotation speeds. WVR (both rotation speeds) had no effect on othlith biogenesis in both stages as all otoliths were present after the experiments. In comparison with 1 g controls, WVR had significantly increased othlith growth (normalised by fish lenth) after 3 and 6 days of exposure, but significant differences of otolith growth between experimental animals and controls were not found after 9 and 12 days. In conclusion, WVR (at least within a time-span of exposure of up to 6 days) brings, comparable to the situation in real microgravity, a kind of feedback mechanism into action, resulting in larger otoliths. Later, possible effects of WVR might be overruled by normal allometric growth since the action of the feedback mechanism may be discontinued in the course of an adaption.