Wu: Pedal Locomotion ABSTRACT Videotapes, both in real time and time lapse, were made of moving anemones stimulated by Aeolidia papillosa. Close analysis of the videotapes provided a detailed descrip¬ tion of the development and progression of locomotory pedal waves in Anthopleura elegantissima. The mean velocity of all anemones studied was 13.6 mm/hr. A correlation between the movements on the column with that of the pedal disc during locomotion provides a basis for the explanation of muscular systems involved. Wu: Pedal Locomotion I. Introduction Although anemones have been known to locomote for well over, a hundred years (Gosse 1860), no detailed analysis of pedal locomotion exists. Furthermore, the majority of the works relating to locomotion have been conducted with the anemone Metridium senile. Almost no work has concentrated on Anthopleura elegantissima except for a brief mention in Edmunds et al (1976). Some of the earlier accounts of pedal locomotion can be found in Parker (1917) for Metridium marginatum, Sagartia luciae, Condylactis passiflora and Actinia bermudensis. Additional accounts using Metridium senile are from Batham and Pantin (1951), Pantin (1952) and Robson (1976). In this study, the pedal locomotion of Anthopleura elegantissima was studied to provide a detailed description of the development and progression of the locomotory waves. Aeolidia papillosa was used as stimulus for the anemones. Videotaping, in both real time and time lapse, was employed as well as simple observation. Questions examined during this study included a detailed description of the mechanics of the pedal waves, the temporal characteristics of the waves and the correlation of movements on the column with that of the pedal disc, to arrive at an explanation of the muscles involved in locomotion. Wu: Pedal Locomotion II. Materials and Methods The experiments were conducted at the Hopkins Marine Station, Pacific Grove and the Monterey Bay Aquarium, Monterey. Both Anthopleura elegantissima and Aeolidia papillosa used were collected from wharf +2 in Monterey. Anemones and nudibranchs were held for approximately seven weeks in 13°c runningsea water. Aeolidia were kept submerged in three feet ofses water to cure and prevent bubble disease. Feeding was intermittent. Only animals that had been without food for at least 24 hrs. were used in experiments. Twenty-four hours without food normally elicits predatory behavior in Aeoldia (cited in Edmunds et al, 1976). Sizes of anemones ranged form 1-3cmacross the oral disc. A. Observation Simple observation of anemones attached to the wall of a glass tank was employed initially. An eolid was introduced until contact was initiated and feeding began. Only eolids of comparable size to anemones were used. Anemone locomotion or detachment would follow. B. Time Lapse Time lapse was conducted at the Monterey Bay Aquarium with a Betacam recorder video camera. Footage was analyzed on a Panasonic Omnivision II VHS videotape system which allowed stop frame, slow speed and normal speed (30 frames/sec) Wu: Pedal Locomotion viewing. One frame was taken every 1.89sec. Filming duration ranged from 1-2hrs. Anemones were attached to the tank wall or a glass plate and then positioned vertically for filming. Filming took place in sea water at room temperature (20°:c) and 14-15°C. Fiber optic lights were employed to reduce heat transfer. All anemones were stimulate by Aeolidia as explained earlier. C. Real Time A Panasonic VHS video recorder was used for filming in real time. Anemones were attached to the vertical walls of small plex¬ iglass tanks. Temperature of the sea water during recording was approximately 17°C. Anemones were stimulated by Aeolidia. D. Locomotion of Anemones Without Oral Disc The oral disc of the anemone was cut off using a pair of surgical scissors. A. elegantissima were allowed to recuperate for approximately 24hrs. Filming conditions were similiar to real time recording. stimulus provided by Aeolidia. E. Quantitative Data Data were gathered from approimately 30 anemones. Not all data were collected from each anemone, that is, only anemone velogity was collected from some anemones while others provide a complete data set. Anemone velocity was measured from the leading edge of the anemone in its original position Wu: Pedal Locomotion to the leading edge of its final position. Wave velocity was measured as the wave travelled across the latter half of the disc where it was easier to follow. Only the number of complete waves crossing thepedal disc were tabulated. Some waves would travel across the disc then die out upon nearing the leading edge. III. Results A. Pedal Wave Pedal locomotion of Anthopleura elegantissima involves monotaxic, direct waves. Waves originate at the point of stimulus and travel across the disc to the leading edge. The posterior portion of the pedal disc is sectioned into lobes by radial furrows running along mesenteric lines. These furrows are areas of contraction and detachment from the substrate (Fig. 1a). As one wave would near completion (Fig. ib). another wave would begin by detaching the back edge and drawing its pedal disc away from the eolid feeding site. Furrows associated with mesentaries begin to deepen and enlarge. As the wave continues to progress, these furrows travel around the edge of the pedal disc becoming a moving wave, and extending toward the center of the disc (Fig. 1c. d). At approximately halfway across the disc (in the direction of locomotion), the furrows become a complete, propagating, direct, monotaxic wave (Fig. le). The wave maintains its Wu: Pedal Locomotion definition as it travels across the remainder of the disc, usually with a slight convexity viewed from the anterior (Fig. 1f,g). Just before the leading edge, the anterior edge of the wave stops and the posterior catches up and forms a crease (Fig. 1h). The column around the leading edge is folded Junder slightly in front of this crease. This folded under portion is flipped out in the final pedal wave sequence. B. Quantitative Data All quantitative data are summarized in Table 1. A similiar "range of variability in each set of data was found for each anemone as well as for the entire experimental group; no conclusive correlation existed between size and velocity, movement forward/wave, etc. C. Movement of the Column during Locomotion Upon initial contact with Aeolidia, Anthopleura would lean towards the eolid with tentacles expanded. But as Aeolidia began feeding, the tentacles were withdrawn slightly and the oral disc contracted down. The anemone soon assumed its original position. The posterior edge inflated and the column moved away from the eolid (Fig. 2a). During the course of a pedal wave the column experiences several peristaltic-like waves passing up and down the column; no conclusive correlations could be made to events in the pedal disc. Wu: Pedal Locomotion Initiation of the wave began as the back edge was simul¬ taneously detached from the substrate and pulled forward toward the leading edge (Fig. 2b). Reattachment occured as the back edge gradually came back into contact with the substrate (Fig. 2c). The wave continues around the posterior pedal disc perimeter as a small fold. Accompanying the appearance of this fold is a lowering of the oral disc located above the wave (Fig. 2d). As the wave becomes a well defined band across the pedal disc, the oral disc above the back edge was raised and a crease was evident in the oral disc (Fig. 2e). Con¬ tinuation of the pedal wave involves a constriction of the lower half of the column, especially near the leading edge (Fig 2f). The anemone begins to lean forward; the column bulges out above the leading edge and gradually makes contact with the substrate (Fig. 2g), folding under in the process. A slight downward contraction of the entire oral disc coincides with this folded pertion being flipped, out as a circumfer¬ ential wave (Fig.2h). IV. Discussion Thus far, the anemone has been Viewed as jeparate elements¬ the pedal disc and column. By correlating the movements of the column to the pedal disc, an attempt at an analysis of the musculature involved is possible. The muscle systems will be generalized from the literature but the studies were largely done one Metridium (Batham and Pantin, 1951). Wu: Pedal Locomotion A. Muscle Systems The two major muscle systems involved in locomotion are the parieto-basilar and endocoelic parietals (Batham and Pantin, 1951). The parieto-basilars provide horizontal to diagonal lines of stress along a mesentery. This muscle system runs from the : pedal disc to the lower half of the body walland are found exocoel¬ ically in perfect and imperfect mesenteries. The parietals provide vertical stress lines within a mesentery. Kunning from the pedal disc to the body wall, parietals exist in the endocoel of microcnemes (Fig.3). Other possible minor contributers are the retractors and the longitudinal muscles of mesenteries (Fig. 4). B. Musculature During Locomotion From my observations,I believe almost all muscle related activity occurs in the mesenteries other than some contraction of the circular muscles of the column. Initiation of a wave is accomplished by the parieto¬ basilars drawing in the back edge while the parietals simul- taneously lift the back portion of the pedal disc off the substrate (Fig. c). The furrows of detachment and contraction that develop are due mainly to the parietals contracting and the parieto¬ basilars contributing (Fig 5d). These furrows are able to propagate around the edge of the disc by progressing from one mesentery to another. Analysis of the Videotapes supports this theory as the furrows move through the visible attachment lines of successive mesenteries Wu: Pedal Locomotion In addition, the creases or lowered oral disc above the wave (Fig2d-f) indicate involvement of the mesenteric longitu - dinal muscles. During a wave, a narrow band of contraction running vertically up the side of the column from the fold at the disc perimeter probably results from both the parietals and parieto-basilars creating a line of tension between column and pedal disc. The appearance of a distinct wave approximately half¬ way across the disc could be due to two perfect mesentaries contracting their parietal muscles; each situated opposite the other along a radial axis (Fig. 5e). For the wave to maintain its definition as it progresses forward through radially arranged mesenteries, isolated con¬ tractions of the fibres in both the parietal and parieto¬ basilar muscle seem necessary (Fig. 5f). These muscle fibers would be in the mesenteries situated at angles perpendicular to the wave. A cinched in contortion appears and passes down the column as the wave approaches the leading edge. This appearance would seem to indicate a contraction of the parieto-basilar muscles propagating the wave across the pedal disc (Fig. 2f, Fig. 5e). As the wave nears competion it becomes a sharp crease, a result of two processes: 1) the excess pedal discmaterial being pushed up against the momentarily stationery leading edge, and 2), the leaning forward and folding under of the column. Wu: Pedal Locomotion 10 Additional observations I made include experiments conducted upon anemones which had their oral discs removed. These anemones, when stimulated by Aeolidia, locomoted normally. This observation agrees with the view that the pedal disc and agacent parts contain all the neuromuscular mechanisms needed for locomotion (Parker, 1917). Decapitation would not affect the parietal and parieto-basilar muscles since they are attached to the body wall. I also noticed a reduction of the pedal disc area before locomoting in anemones which had their pedal discs spread out. Waves would be initiated and propagate as far as the front portion of the pedal disc but then stopped. This is probably associated with contraction of the circular muscles of the pedal disc to reduce its area. Further work is clearly indicated to clarify the two previously mentioned behavioral responses. A detailed analysis of the musculature of A. elegantissima especially during lo¬ comotion would greatly contribute to our understanding of pedal locomotion. Questions such as, why is the manner of propagation seemingly different in the beginning compared to the end of a wave and, what manner of nervous activity is involved, remain unanswered. edal moti I would like to thank everyone at Hopkins Marine Station, particularly Freya Sommers, and Chuck Baxter for his invaluable guidance and assistance. A special thanks to Mark Shelley and Alex Edwards of Monterey Bay Aquarium for their time, energy and generosity in allowing me to use the time lapse photography equipment. Wu: Pedal Locomotion 12 REFERENCES Batham, E.J. and Pantin, C.F.A. 1950 Muscular and Hydrostatic Action in the Sea Anemones Metridium senile (L.) Journ. of Ex. Biol. 27:264-289. Batham, E.J., and Pantin, C.F.A. 1951 The Organization of the Muscular System of Metridium senile. Quart. Journ. Micro. Sci. 92:27-54. Edmunds, M. et al 1976 Defensive Behavior of Sea Anemones in Response to Predation by the Opisthobranch Mollusc Aeolidia papillosa (L.). J. Mar. Biol. Ass. U.K., 56:65-83. Pantin, C.F.A. 1952 The Elementary Nervous System. Proc. Roy. Soc. B.140:147-168. Parker, G.H. 1917 Pedal Locomotion in Actinians. Journ. of Exp. Zool., 22:111-124. Robson, E.A. 1976 Locomotion in Sea Anemones: The Pedal Disk, In Coelenterate Ecology and Behavior (ed G.O. Mackie), Plenum Press. New York and London. Wu: Pedal Locomotion FIGURE CAPTIONS Figure 1: Sequence of pedal wave outlines during locomotion. The two major muscle systems associated with Figure 2: locomotion. Taken from Batham and Pantin (1951) for Metridium senile. Figure 3: Musculature of a perfect mesentery. Other muscle systems which may have minor involvement in locomotion-- longitudinal, retractors and circular. Taken from Batham and Pantin (1951) for Metridium senile. Figure 4: Movement of the column during locomotion as it relates to the pedal wave. Anemone was stimulated by Aeolidia papillosa. Figure 5: Mesentery musculature during locomotion. a) Top viey of mesenteries. b) Cross-section of a mesentery, c) wave beginning as shown in one mesentery. Parietals and parieto-basilars contract. d) Deepening furrows through contraction of parietals and parieto-basilar. e) View of one half of pedal disc. Perfect mesentery contracting muscies along vertical axis. f) Section of pedal disc showing three simplified mesenteries. g) Cross section of mesentery with edge of column folded under. Contraction of parieto-basilars combined with hydroståtic pressure flip folded under portion back out. 1 3 9 9 —-- W ( ( O (C ORAL STOMA RETRACTOR LONGITUDINAL RADIAL PARIETO-BASILAR DISK CIRCULAR 4 szasse 11 — (C (c) (a) Se k (f) (d) A (g) (b) (e) ——— „— — . PARIETAL r PARIETO-BASILAR RETRACTOR -- CIRCULAR¬ C Wu: Peda comotion TABLE CAPTIONS Table 1: Anemone and wave velocities of A. elegantissima in response to an attack by Aeolidia papillosa. 14 2 NO — 1N O L I LL —:— LE 2 — â — AT E