Na⁺/K⁺-ATPase (NKA) function is inhibited by Bufadienolides (BD), a group of cardiotonic steroids (CTS) primarily produced by anurans of the Bufonidae family, such as Rhinella marina. This study characterized the presence of α and β NKA subunit isoforms in R. marina via RNAseq in four tissues: oocytes, skin, heart, and skeletal muscle. Transcripts encoding three α-like isoforms (α₁, α₂, α₃) and three β-like isoforms (β₁, β₂, β₄) were identified. The amino acid sequence of α₁-like isoform shared 99.4% identity with the α₁ isoform previously published for R. marina. Sequences for α₂, α₃, and β₄ from R. marina were previously unavailable. The first extracellular loop in the α₂-like isoform in R. marina showed similar substitutions to those found in their susceptible homologues in other taxa (L/Q111T and S119T); in contrast, this same loop in α₃-like isoform showed similar substitutions (Q111L and G120R) to those reported for toad-eating animals such as snakes, which suggests relatively lower affinity for CTS. Docking results showed that all three α-like isoforms identified in R. marina transcriptomes have low affinity to CTS compared to the susceptible α₁ isoform of Sus scrofa (pig), with α₁-like isoform being the most resistant. The tissue-specific RNAseq results showed the following expression of NKA α-like and β-like subunit isoforms: Oocytes expressed α₁ and β₁; skin α₁, β₁, and low levels of β₂; heart α₁, α₃, and β₁; skeletal muscle α₁, β₄, with low levels of α₂, α₃, and β₁. R. marina could be used as an important model for future structural, functional and pharmacological studies of NKA and its isoforms.

的Na ⁺ / K ⁺ -ATP酶（NKA）功能由Bufadienolides（BD），一组强心类固醇（CTS）主要由蟾蜍家庭的无尾两栖类，如产生的抑制Rhinella码头。本研究通过 RNAseq 在卵母细胞、皮肤、心脏和骨骼肌这四种组织中表征了R. marina中 α 和 β NKA亚基亚型的存在。鉴定了编码三种 α 样同种型（α ₁、α ₂、α ₃）和三种 β 样同种型（β ₁、β ₂、β ₄）的转录本。的α的氨基酸序列₁样同种型共享99.4％的同一性与α ₁先前为R. marina发表的同种型。以前无法获得来自R. marina 的α ₂、α ₃和 β ₄序列。R. marina 中α ₂样亚型中的第一个细胞外环显示出与其他类群（L/Q111T 和 S119T）中易感同源物相似的替换；相比之下，α ₃样亚型中的相同环显示出与蛇等食蟾蜍动物报道的相似替代（Q111L 和 G120R），这表明对 CTS 的亲和力相对较低。对接结果表明，在R. marina 中鉴定出的所有三种 α 样亚型与Sus scrofa（猪）的易感 α ₁同种型相比，转录组对 CTS 的亲和力较低，其中 α ₁样同种型最具抗性。组织特异性 RNAseq 结果显示 NKA α-like 和 β-like 亚基亚型的表达如下： 卵母细胞表达 α ₁和 β ₁；皮肤α ₁、β ₁和低水平的β ₂；心脏α ₁、α ₃和β ₁；骨骼肌α ₁、β ₄，α ₂、α ₃和β ₁水平较低。玛丽娜河 可用作未来 NKA 及其同种型的结构、功能和药理学研究的重要模型。