A Stimulus Paradigm for Analysis of Near-field Hydrodynamic Sensitivity in Crustaceans

We present several relatively simple procedures for studying the physiology of near-field mechanoreceptors in crustaceans which extend previous measures of sensitivity. The advantages include the quantitative analysis of range fractionation and directionality of receptors and interneurons in the sensory hierarchy of the central nervous system (CNS), based on a stimulus paradigm that is reproducible and easy to use. The technical considerations for quantitative fluid-coupled stimulation addressed by this paper are the complexity of dipole flow fields, reflected interference from traveling waves, and the underlying stimulus wave form. The techniques described here offer corresponding advantages for physiological experiments using other aquatic organisms. In electrophysiological experiments, crustacean preparations are typically placed in an experimental chamber filled with water or saline solution. For studies on near-field sensory receptors, i.e. those responding to flow fields in the aquatic medium, a dipole or vibrating sphere is frequently used to generate stimulus waves (Tautz et al. 1981; Wiese and Wollnik, 1983; Ebina and Wiese, 1984; Hatt, 1986; Heinisch and Wiese, 1987; Tautz, 1987; Wiese and Marschall, 1990; Killian and Page, 1992b). A dipole stimulator is easily constructed by attaching a spherical probe to an electromechanical device such as a loudspeaker, pen motor or piezo crystal. A periodic signal fed to the transducer generates the oscillating dipole movements. With the sphere immersed in the bathing medium, dipole flow fields are generated (see Kalmijn, 1988, for further discussion of dipole sources), whereas dipole oscillations introduced at the air­saline interface generate traveling surface waves. Numerous additional devices and techniques have been used to stimulate crustacean receptors. Several involve wave motion introduced from one end of the chamber by diaphragms or paddles (Laverack, 1962b, 1963; Flood and Wilkens, 1978), by cylindrical dippers (Wilkens and Larimer, 1972; Wiese et al. 1976; Wiese and Schultz, 1982; Reichert et al. 1983) or by water drops (Laverack, 1962b; Strandburg and Krasne, 1985). Another form of fluid-coupled stimulation involves small jets of saline (Laverack, 1963; Tautz, 1990; Schmitz, 1992). In other studies, receptor hairs have been stimulated directly by using a stylus in place of the dipole, e.g. a needle or glass capillary in contact with the hair (Laverack, 1962a; Killian and Page, 1992a; Yen et al. 1992; Nagayama and Sato, 1993) or a miniature wire loop or capillary tube placed over the hair shaft (Mellon, 1963; Wiese, 1976; Tautz et al. 1981; Killian and Page, 1992b).