Martinez Nathan P, Pinch Matthew, Kandel Yashoda, Hansen Immo A
Department of Biology, New Mexico State University, Las Cruces, NM 88003, USA.
Insects. 2023 Jan 5;14(1):50. doi: 10.3390/insects14010050.
The Na/K ATPase (NKA) is present in the cellular membrane of most eukaryotic cells. It utilizes energy released by ATP hydrolysis to pump sodium ions out of the cell and potassium ions into the cell, which establishes and controls ion gradients. Functional NKA pumps consist of three subunits, alpha, beta, and FXYD. The alpha subunit serves as the catalytic subunit while the beta and FXYD subunits regulate the proper folding and localization, and ion affinity of the alpha subunit, respectively. Here we demonstrate that knockdown of NKA beta subunit 2 mRNA ( reduces fecundity in female . We determined the expression pattern of in several adult mosquito organs using qRT-PCR. We performed RNAi-mediated knockdown of and assayed for lethality, and effects on female fecundity. Tissue expression levels of mRNA were highest in the ovaries with the fat body, midgut and thorax having similar expression levels, while Malpighian tubules had significantly lower expression. Survival curves recorded post dsRNA injection showed a non-significant decrease in survival of dsRNA-injected mosquitoes compared to GFP dsRNA-injected mosquitoes. We observed a significant reduction in the number of eggs laid by dsRNA-injected mosquitoes compared to control mosquitoes. These results, coupled with the tissue expression profile of indicate that this subunit plays a role in normal female fecundity. Additional research needs to be conducted to determine the exact role played by NKAβ2 in mosquito post-blood meal nutrient sensing, transport, yolk precursor protein (YPP) synthesis and yolk deposition.
钠钾ATP酶（NKA）存在于大多数真核细胞的细胞膜中。它利用ATP水解释放的能量将钠离子泵出细胞，将钾离子泵入细胞，从而建立并控制离子梯度。功能性NKA泵由α、β和FXYD三个亚基组成。α亚基作为催化亚基，而β和FXYD亚基分别调节α亚基的正确折叠和定位以及离子亲和力。在这里，我们证明敲低NKAβ2亚基mRNA会降低雌性的繁殖力。我们使用qRT-PCR确定了几种成年蚊虫器官中的表达模式。我们进行了RNAi介导的敲低，并检测了其致死率以及对雌性繁殖力的影响。NKAβ2 mRNA的组织表达水平在卵巢中最高，脂肪体、中肠和胸部的表达水平相似，而马氏管的表达水平明显较低。注射dsRNA后记录的存活曲线显示，与注射GFP dsRNA的蚊子相比，注射NKAβ2 dsRNA的蚊子的存活率没有显著下降。与对照蚊子相比，我们观察到注射NKAβ2 dsRNA的蚊子产卵数量显著减少。这些结果，再加上NKAβ2的组织表达谱，表明该亚基在正常雌性繁殖力中发挥作用。需要进行更多研究以确定NKAβ2在蚊虫吸血后营养感知、运输、卵黄前体蛋白（YPP）合成和卵黄沉积中的确切作用。
