Leibniz Institute of Marine Sciences, Kiel, Germany.
Cell Tissue Res. 2010 Mar;339(3):571-83. doi: 10.1007/s00441-009-0921-8. Epub 2010 Feb 2.
The tissue distribution and ontogeny of Na(+)/K(+)-ATPase has been examined as an indicator for ion-regulatory epithelia in whole animal sections of embryos and hatchlings of two cephalopod species: the squid Loligo vulgaris and the cuttlefish Sepia officinalis. This is the first report of the immunohistochemical localization of cephalopod Na(+)/K(+)-ATPase with the polyclonal antibody alpha (H-300) raised against the human alpha1-subunit of Na(+)/K(+)-ATPase. Na(+)/K(+)-ATPase immunoreactivity was observed in several tissues (gills, pancreatic appendages, nerves), exclusively located in baso-lateral membranes lining blood sinuses. Furthermore, large single cells in the gill of adult L. vulgaris specimens closely resembled Na(+)/K(+)-ATPase-rich cells described in fish. Immunohistochemical observations indicated that the amount and distribution of Na(+)/K(+)-ATPase in late cuttlefish embryos was similar to that found in juvenile and adult stages. The ion-regulatory epithelia (e.g., gills, excretory organs) of the squid embryos and paralarvae exhibited less differentiation than adults. Na(+)/K(+)-ATPase activities for whole animals were higher in hatchlings of S. officinalis (157.0 +/- 32.4 micromol g (FM) (-1) h(-1)) than in those of L. vulgaris (31.8 +/- 3.3 micromol g (FM) (-1) h(-1)). S. officinalis gills and pancreatic appendages achieved activities of 94.8 +/- 18.5 and 421.8 +/- 102.3 micromol(ATP) g (FM) (-1) h(-1), respectively. High concentrations of Na(+)/K(+)-ATPase in late cephalopod embryos might be important in coping with the challenging abiotic conditions (low pH, high pCO(2)) that these organisms encounter inside their eggs. Our results also suggest a higher sensitivity of squid vs. cuttlefish embryos to environmental acid-base disturbances.
我们通过免疫组织化学方法，利用针对人源 Na(+)/K(+)-ATP 酶α1 亚基的多克隆抗体 alpha (H-300)，对两种头足类动物（鱿鱼 Loligo vulgaris 和乌贼 Sepia officinalis）的胚胎和幼体的整体动物切片进行研究，以检测 Na(+)/K(+)-ATP 酶的组织分布和个体发生情况。这是首次报道使用针对人源 Na(+)/K(+)-ATP 酶α1 亚基的多克隆抗体 alpha (H-300)对头足类动物的 Na(+)/K(+)-ATP 酶进行免疫组织化学定位。在几种组织（鳃、胰腺附属物、神经）中观察到 Na(+)/K(+)-ATP 酶的免疫反应性，这些组织的免疫反应性仅局限于血液窦的基底外侧膜。此外，在成年鱿鱼 L. vulgaris 标本的鳃中，观察到大型单个细胞类似于在鱼类中发现的富含 Na(+)/K(+)-ATP 酶的细胞。免疫组织化学观察表明，晚期乌贼胚胎的 Na(+)/K(+)-ATP 酶的数量和分布与幼体和成年阶段相似。鱿鱼胚胎和幼体的离子调节上皮（例如鳃、排泄器官）的分化程度低于成年个体。乌贼 S. officinalis 幼体的 Na(+)/K(+)-ATP 酶活性（157.0 +/- 32.4 μmol g (FM) (-1) h(-1)）高于鱿鱼 L. vulgaris 幼体（31.8 +/- 3.3 μmol g (FM) (-1) h(-1)）。乌贼的鳃和胰腺附属物的 ATP 酶活性分别达到 94.8 +/- 18.5 和 421.8 +/- 102.3 μmol(ATP) g (FM) (-1) h(-1)。在这些生物体的卵内，晚期头足类胚胎中高浓度的 Na(+)/K(+)-ATP 酶可能对于应对具有挑战性的非生物条件（低 pH 值、高 pCO(2)）非常重要。我们的研究结果还表明，鱿鱼胚胎比乌贼胚胎对环境酸碱干扰更敏感。
