POWRE: The Evolving Function of the Basolateral Membrane Ca Pump in Exchange Epithelia of a Continuum of Aquatic to Semi-Terrestrial Brazilian Crabs

小麦9870374 这笔拨款将使PI能够评估钙转运蛋白是如何进化的。 PI将研究钙泵在运输上皮细胞（鳃，肾脏，肠道）的生理功能和分子特征，巴西现有的相关水生到半陆生螃蟹的进化连续体。 甲壳类动物被用作研究钙平衡的非哺乳动物模型系统，这是由于伴随其自然蜕皮周期的矿化/脱矿过程。 该假说是，在从水到陆地的进化过程中，水生钙吸收模式（通过鳃）被陆地模式（通过肠道的饮食摄入）所取代。 具体的目标是检查钙吸收蛋白的鳃，肾脏和肠道在每只螃蟹的功能（a）在蜕皮周期的阶段和（B）CA的可用性（水或饮食）。 方法将包括制作膜囊泡（类似于充满水的气球），以研究体外钙摄取以及使用分子生物学克隆和测序组织特异性基因。 将在蜕皮期间定量基因的表达并作为Ca可用性的函数。 钙泵是所有真核细胞中钙稳态的基础。 这项研究将描述这种重要分子在过去6亿年中的演变。

Wheatly 9870374 The grant will allow the PIs to assess how calcium transporting proteins evolved. The PIs will study the physiological function and molecular characterization of the calcium pump in transporting epithelia (gills, kidney, gut) of an evolutionary continuum of related aquatic to semi-terrestrial crabs available in Brazil. Crustaceans are used as a non-mammalian model system for the study of calcium balance due to the mineralization/demineralization processes that accompany their natural molting cycle. The hypothesis is that during evolution from water onto land aquatic modes of Calcium uptake (across gills) become superseded by terrestrial modes (dietary intake across gut). The specific objectives are to examine the Ca uptake proteins in the membrane of gill, kidney and gut in each crab as a function of (a) stage in the molting cycle and (b) CA availability (water or dietary). Methods will include making membrane vesicles (analogous to water filled balloons) to study Ca uptake in vitro as well as using molecular biology to clone and sequence tissue-specific genes. Expression of the genes will be quantified during molting and as a function of Ca availability. The Ca pump is fundamental to Ca homeostasis in all eukaryotic cells. This research will delineate the evolution of this critical molecule over the past 600 million years.