Heat shock response and shape regulation during newt tail regeneration

Regenerating newt tail has recently been found to react to hypergravity in a stable and reproducible way by curving downwards. Such morphogenetic effect of non-specific physical factor applied to a complex structure of an adult animal is a rare phenomenon with unknown molecular basis. For the first steps of unraveling this basis we've chosen heat shock proteins (HSPs) as promising candidates. Morphometrical analysis of tail regeneration was performed in aquarium (control), on substrate (relative hypergravity) and in aquarium under weekly application of heat shock. HSPs were inhibited pharmacologically during regeneration in aquarium and on substrate. Hsp70, 90 gene expression and protein localization were analyzed in the studied conditions. Weekly application of heat shock to newts regenerating tails in otherwise normal conditions led to development of curved tails (both upwards and downwards), suggesting that similar mechanisms are at play in both hyper gravity-altered and heat shock-altered morphogenesis. Heat shock protein inhibitor KNK437 didn't affect tail shape during normal regeneration, but prevented the formation of tail curve in appropriate conditions. It was shown that HSP70 and HSP90 proteins are present in muscle and connective tissue of intact tails as well as regenerates, but only appear in epidermis in hypergravity-altered regenerates and heated tails. Based on our data, we hypothesize, that different external factors (e.g. hypergravity and heat shock) are received, analyzed and transmitted further to affect morphogenesis by similar mechanisms that utilize a set of HSP in epidermal cells.