Department of Biological Sciences, University of Cincinnati, Cincinnati, OH 45221-0006, USA.
J Exp Biol. 2011 Nov 1;214(Pt 21):3524-31. doi: 10.1242/jeb.058990.
The principal eyes of sunburst diving beetle, Thermonectus marmoratus, larvae are among the most unusual eyes in the animal kingdom. They are composed of long tubes connecting bifocal lenses with two retinas: a distal retina situated a few hundred micrometers behind the lens, and a proximal retina that is situated directly beneath. A recent molecular study on first instar larvae suggests that the distal retina expresses a long-wavelength-sensitive opsin (TmLW), whereas the proximal retina predominantly expresses an ultraviolet-sensitive opsin (TmUV II). Using cloning and in situ hybridization we here confirm that this opsin distribution is, for the most part, maintained in third instar larvae (with the exception of the TmUV I that is weakly expressed only in proximal retinas of first instar larvae). We furthermore use intracellular electrophysiological recordings and neurobiotin injections to determine the spectral sensitivity of individual photoreceptor cells. We find that photoreceptors of the proximal retina have a sensitivity curve that peaks at 374-375 nm. The shape of the curve is consistent with the predicted absorbance of a single-opsin template. The spectral response of photoreceptors from the distal retina confirms their maximum sensitivity to green light with the dominant λ-peak between 520 and 540 nm, and the secondary β-peak between 340 and 360 nm. These physiological measurements support molecular predictions and represent important steps towards understanding the functional organization of the unusual stemmata of T. marmoratus larvae.
日射甲虫幼虫的主眼是动物王国中最不寻常的眼睛之一。它们由连接双焦点透镜和两个视网膜的长管组成:一个位于透镜后几百微米处的远端视网膜,和一个直接位于下方的近端视网膜。最近对第一龄幼虫的分子研究表明,远端视网膜表达一种长波敏感视蛋白(TmLW),而近端视网膜主要表达一种紫外线敏感视蛋白(TmUV II)。使用克隆和原位杂交,我们在这里证实,这种视蛋白的分布在第三龄幼虫中基本保持不变(除了 TmUV I,它仅在第一龄幼虫的近端视网膜中微弱表达)。我们还使用细胞内电生理记录和神经生物素注射来确定单个光感受器细胞的光谱敏感性。我们发现,近端视网膜的光感受器的敏感性曲线在 374-375nm 处达到峰值。该曲线的形状与单一视蛋白模板的预测吸收率一致。来自远端视网膜的光感受器的光谱响应证实了它们对绿光的最大敏感性,主要 λ-峰值在 520nm 到 540nm 之间,次要 β-峰值在 340nm 到 360nm 之间。这些生理测量结果支持分子预测,并代表了理解 T. marmoratus 幼虫不寻常的眼点的功能组织的重要步骤。