Janahmadi M, Malmierca M S, Hearne P G, Green G G, Sanders D J
Department of Physiological Sciences, The Medical School, University of Newcastle upon Tyne, UK.
Anat Embryol (Berl). 1999 Jun;199(6):563-72. doi: 10.1007/s004290050253.
Identified neurones F76 and D1 of the suboesophageal ganglia of Helix aspersa were studied in the isolated ganglia in vitro and in culture. The neurones were examined electrophysiologically with current clamp and morphologically either with intracellular injections of Lucifer Yellow or biocytin. These nerve cells had very similar resting membrane potentials and responses to injected current. The projections of D1 and F76 have been characterised, with both neurones having two main axons. The F76 neurones project to the left pallial, right pallial, anal, and visceral nerves as well as to the left and right pleural ganglia. The D1 neurones have similar projections except that they do not project to the anal and visceral nerves. The bilateral symmetry to the pallial nerves and pleural ganglia is discussed. These cells were also studied electrophysiologically after mechanical isolation and culture. F76 and D1 neurones were separated by dissection (no enzymes) and cultured in three ways. In normal snail Ringer they remained viable for up to two weeks with no development. In Ringer preincubated with a ganglia or containing endothelial growth factor, neurite outgrowths were seen. Membrane potentials were significantly lower in cultured neurones than in vitro and the after hyperpolarization never went below resting in cultured cells but it did in vitro.
对散大蜗牛食管下神经节中已识别的神经元F76和D1进行了体外分离神经节和培养研究。采用电流钳对神经元进行电生理检查，并用荧光黄或生物素细胞内注射进行形态学检查。这些神经细胞具有非常相似的静息膜电位和对注入电流的反应。已对D1和F76的投射进行了表征，这两种神经元都有两条主要轴突。F76神经元投射到左外套膜、右外套膜、肛门和内脏神经以及左、右胸膜神经节。D1神经元有类似的投射，但不投射到肛门和内脏神经。讨论了对外套膜神经和胸膜神经节的双侧对称性。在机械分离和培养后也对这些细胞进行了电生理研究。通过解剖（不使用酶）分离F76和D1神经元，并以三种方式进行培养。在正常蜗牛林格液中，它们可存活长达两周且无发育。在预先用神经节孵育或含有内皮生长因子的林格液中，可观察到神经突生长。培养神经元的膜电位显著低于体外，且培养细胞的超极化后电位从未低于静息电位，但在体外会低于静息电位。
