Takigami Satoshi, Sunada Hiroshi, Horikoshi Tetsuro, Sakakibara Manabu
Graduate School of High-Technology for Human Welfare, Tokai University, Numazu, Shizuoka 410-0321, Japan; Graduate School of Bioscience, Tokai University, Numazu, Shizuoka 410-0321, Japan.
Hotchkiss Brain Institute, Faculty of Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada.
Biophysics (Nagoya-shi). 2014 Nov 11;10:77-88. doi: 10.2142/biophysics.10.77. eCollection 2014.
Dermal photoreceptors located in the mantle of Lymnaea stagnalis were histologically and physiologically characterized. Our previous study demonstrated that the shadow response from dermal photoreceptors induces the whole-body withdrawal response. Through the interneuron, RPeD11, we detected that the light-off response indirectly originated from a dermal photoreceptor. Previous observations, based on behavioral pharmacology, revealed that cyclic guanosine monophosphate acts as a second messenger in the dermal photoreceptor. Furthermore, gastropods possess dermal photoreceptors containing rhodopsin, as a photopigment, and another photo-sensitive protein, arrestin, responsible for terminating the light response. Thus, we chose three antibodies, anti-cGMP, anti-rhodopsin, and anti-β-arrestin, to identify the dermal photoreceptor molecules in Lymnaea mantle. Extracellular recording, using a suction electrode on the mantle, revealed a light off-response from the right parietal nerve. Overlapping structures, positive against each of the antibodies, were also observed. Numerous round, granular particles of 3-47 μm in diameter with one nucleus were distributed around pneumostome and/or inside the mantle. The cells surrounding the pneumostome area, located 10 μm beneath the surface, tended to have smaller cell soma ranging from 3 to 25 μm in diameter, while cells located in other areas were distributed uniformly inside the mantle, with a larger diameter ranging from 12 to 47 μm. The histological examination using back-filing Lucifer Yellow staining of the right parietal nerve with the three dermal photoreceptor antibodies confirmed that these overlapping-stained structures were dermal photoreceptors in Lymnaea.
对椎实螺(Lymnaea stagnalis)外套膜中的皮肤光感受器进行了组织学和生理学特征分析。我们之前的研究表明,皮肤光感受器的阴影反应会诱发全身退缩反应。通过中间神经元RPeD11,我们检测到熄灯反应间接源自皮肤光感受器。基于行为药理学的先前观察表明,环磷酸鸟苷在皮肤光感受器中充当第二信使。此外,腹足纲动物拥有含有视紫红质(作为一种光色素)和另一种负责终止光反应的感光蛋白——抑制蛋白的皮肤光感受器。因此,我们选择了三种抗体,即抗环磷酸鸟苷抗体、抗视紫红质抗体和抗β - 抑制蛋白抗体,来鉴定椎实螺外套膜中的皮肤光感受器分子。使用外套膜上的吸电极进行细胞外记录,揭示了右顶神经的熄灯反应。还观察到了对每种抗体呈阳性的重叠结构。许多直径为3 - 47μm、带有一个细胞核的圆形颗粒状物质分布在气门周围和/或外套膜内部。位于表面下方10μm处的气门区域周围的细胞,其细胞体往往较小,直径在3 - 25μm之间,而位于其他区域的细胞则均匀分布在外套膜内,直径较大,在12 - 47μm之间。使用三种皮肤光感受器抗体对右顶神经进行回注荧光黄染色的组织学检查证实,这些重叠染色的结构是椎实螺的皮肤光感受器。