Thermal desiccation and relative gene expression of HSP90 in an acorn barnacle, Amphibalanus amphitrite
Abstract
Sessile and sedentary invertebrates respond to various magnitudes of thermally desiccated stress during tidal emersion, and regulation of body temperature plays a crucial role in survival during an intraday aerial exposure. Survival strategies often include ecological and molecular adaptation strategies such as modifying body temperature (shell temperature) and expression of stress genes such as Heat shock protein 90 (HSP90). Estimation of gene expression through real-time qPCR is ideal for unravelling the intricate molecular mechanisms underlying the stress adaptation of intertidal invertebrates. Selection of reference genes (RGs) before qPCR experiments requires a systematic assessment of RGs relating to the adaptation of the intertidal barnacles. Marine intertidal barnacles such as Amphibalanus amphitrite are infamous crustaceans from the perspective of biofouling and antifouling. They are extensively studied for their larval ecology, stress adaptation, settlement and recruitment, etc. In this study, we implemented RefFinder tool to evaluate the expression stability of RGs such as Actin muscle (ACT), 18S rRNA aminocarboxypropyltransferase-like (18S rRNA), 28S rRNA, Tubulin (TUB), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and Actin alpha (ACT-A) in adult A. amphitrite. This study is the first of its kind to investigate and validate the RGs (ACT-A and GAPDH) towards Relative Gene Expression (REGs) (fold change) using stress gene hsp90-CDC37like (HSP90). The findings of this study indicated that ACT-A and GAPDH are the most stable and ideal reference genes to perform relative gene expression (REGs) of thermally desiccated A. amphitrite. Additionally, stable RGs were validated for relative gene expression of stress gene HSP90 among thermally-desiccated A. amphitrite. An upregulation of HSP90 gene expression in barnacles upon 2h of thermal-desiccation was observed, which can be attributed to subtle rise in body temperature by 0.5��C during early emersion.
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