Lee: Independent catch tentacle TRACT The inflated catch tentacle of Metridium senile behaves independent of the oral disk. Close obser- vation during aggression toward Anthopleura elegantissima reveals clues to its hydrostatic and neural separation from the rest of the animal. In the development of the catch tentacle from regular tentacle, changes in cnidom, musculature, and nerve net are necessary to make it a functional unit. Lee: Independent catch tentacle INTRODUCTION The movements of specialized, highly inflatable tentacles in sea anemones were first described by Gosse (1860). Hand (1955) described catch tentacles as occurring in one percent of Metridium senile in Central California. Catch tentacles have been identified in six families of the acontiate anemones: Diadumenidae, Sagartiidae, Metridiidae, Isopheliidae, Sagartiomorphidae, and Haliplanellidae (Williams, 1975). In Metridium catch tentacles are used in both intra- specific and interspecific aggression (Purcell, 1977). Purcell asserted that aggressive Metridium on the edges of adjacent clones maintained anemone-free zones. More recently these zones have disappeared and clones are no longer segregated (Purcell, 1982). A similar aggressive behavior was also observed in Anthopleura elegantissima (Francis, 1973). Catch tentacles can be induced after prolonged tentacle contact with non-clonemates. After three weeks catch tentacles were inflatable, and after nine weeks they were fully differentiated (Purcell, 1977). This study aimsto analyze the behavior pattern of an inflatable catch tentacle, and the control of its response and movement. Catch tentacles represent an inducible developmental system which exhibits a Lee: Independent catch tentacle complex and prolonged endogenous behavior pattern released by an appropriate stimulus. MATERIALS AND METHODS Several clones of Metridium senile were collected from the pilings at Wharf No. 2 in Monterey Bay. Size Tranged from 5 to 7 cm in column height and 4 to 6 cm across the oral disk. Anthopleura elegantissima were collected from the intertidal at Hopkins Marine Station in Pacific Grove. They were approximately 4 om across the oral disk. Separate tanks with running sea water at 14 C held the two anemones. They were settled on movable rocks. Every 5 days they were fed Tegriopus. To induce catch tentacle inflation Metridium was placed in tentacle tip contact with Anthopleura. This constituted the start of stimulus, and responses were timed from this point with a watch. The catch tentacle need not be fully inflated to count as a response, but it must be markedly distended. Catch tentacles around the oral disk were divided into six 60ssectors. Each time the point of stimulus became the center of sector one. For microscopy a whole Metridium was relaxed in 7% MgCl, sea water 1:1 for over 14 hours. Tentacles were transected at the base and cut open to expose Lee: Independent catch tentacle the endoderm. Videotape was utilized to analyze aspects of Metridium catch tentacle behavior. RESULTS AND DISCUSSION The typical action pattern of a catch tentacle is described and illustrated in Figure 1. (1) Before stimulation a catch tentacle is still and unexpanded. It is more opaque than the feeding tentacles. (2) At initial contact with Anthopleura nearby tentacles contract, and if the stimulus was large the oral dise will retract. (3) After a variable lag period catch tentacle begins to elongate. This is caused by increase in hydrostatic pressure, and maybe contraction of muscle in the tentacle. (4) Inflation starts at the base and moves up to the tip. It is not a continuous process because the catch tentacle repeatedly contracts and expands. It can inflate to a maximum size of 0.5cm x 12 cm. (5) The movements of the tentacle originate at the base and transmit along the whole length. It is analogous to the motion of a whip. Muscular action move the inflated tentacle all around the oral disc. Lee: Independent catch tentacle (6) The tentacle tip attaches anywhere on Anthopleura. Nematocysts anchor it and then theet Metridium contract and break the tip. The aggressive behavior of Metridium was fairly consistent. The response of catch tentacles followed tentacle contact with Anthopleura. Catch tentacles were inflated in 37/52 trials, or 71%. The response times of this group varied greatly (Figure 2). In this data the duration of stimulation (the time the anemones were left together) varied from ten seconds to the length of the trial. If the time parameter was held constant at ten minutes, the lag period maintained its high variability (Table 1). The catch tentacle to inflate first was usually the one closest to the site of stimulus (Figure 3). Of the 37 anemones that responded, 21 or 51% inflated more than one catch tentacle. This would occur within a period of five minutes. An inflated catch tentacle seems to be hydro¬ statically isolated from the rest of the coelenteron. When red food color was injected into the column of an anemone with inflated catch tentacles, no color appeared in the coelenteron of the catch tentacles. The color diffused to all other parts of the animal. Dye injected to an unstimulated Metridium was dis- Lee: Independent catch tentacle charged before catch tentacles could be induced to inflate. Methylene blue, carmine, and graphite particles gave similar results. Examination of the oral disc revealed that catch tentacles occurred only between pairs of perfect mesenteries (Table 2). Under the base of an inflated tentacle one could see a mesentery pair coming together, creating a small compartment. If the mesenteric stoma closes and the circular and radial muscles of that endocoel contract, that would isolate the endocoel from the rest of the coelenteron (Figure 4). Movements of the catch tentacle did not affect the appearance or activity of feeding tentacles. A Metridium often inflated 2 to 6 tentacles without effect on the oral disc. Touching an inflated tentacle caused it to retract, but with no reaction around the oral disc. All the observations together suggest a separate hydrostatic and neural control of the catch tentacle from the oral disc and regular tentacles. Catch tentacles and regular tentacles were anaesthetized, slit longitudinally, and prepared as whole mounts for viewing. Figure 5 shows the longitudinal muscle fibers of a regular tentacle, The neuromuscular structure of Metridium tentacles Lee: Independent catch tentacle have been described by van Marle (1976), and only longitudinal fibers were mentioned. Batham and Pantin (1951) described, in addition, circular endodermal muscles, which were not observed in this study. The catch tentacles have cross fibers oriented at 90° (Figure 6). Higher magnification (1000 x) revealed spindle-shaped cells. The thicker, longitudinal fibers were closer to the epithelium and the cross fibers were just above them. In swimming sea anemone Gonactinia prolifera, tentacles were used for walkingi and swimming. The tentacles had ectododermal longi- tudinal fibers and endodermal circular fibers (Robson, 1971). Although the activity of catch tentacle is not as strenuous as walking and swimming, the tissue undergoes frequent expansion and retrac¬ tion. The crossing muscular system also provides a full range of motion in its aggressive behavior. GENERAL DISCUSSION The catch tentacle of the clonally reproducing Metridium senile represents a behavioral system with some interesting properties. A simple structure with complex behavior can be developmentally induced from a regular tentacle, and it can regress to its original structure (Purcell, 1977, Hand, 1955). Lee: Independent catch tentacle The nerve nets of coelenterates have received con¬ siderable attention, and a picture of separate conducting systems controlling a variety of behaviors with varying degrees of linkage between these systems is emerging. The nerve net of Metridium has been characterized for the mesentery and column (Batham, et al, 1960, Robson and Josephson, 1969). The neural control of the catch tentacle adds another layer to this with a system having interesting and tractable properties. Spread of excitation from the stimulus site, tip of a regular tentacle, down to the oral disc and to the catch tentacles takes much longer than expected for conduction in a nerve net. The wide range of response times of Metridium to ag- gressive stimulus make speculation on the speed of this conduction difficult. Probably there is some utilization time to release the endogenous mechanism. In any case the nervous control of the catch tentacle appears to be independent of the general nerve net. The feeding tentacles resume normal activities after initial contact with Anthopleura, and lack remark- able movement during catch tentacle aggression. A large, noxious stimulus such as a hypodermic injection causes overall contraction and retraction of inflated catch tentacle. Five or six inflated Lee: Independent catch tentacle tentacles of a stimulated anemone gradually move toward the site of stimulus. However there is lack of coordinated movement among these tentacles. Thus each tentacle can exhibit spatial orientation toward the site of stimulation, though it is removed from that sector. Catch tentacles can be induced from feeding tentacles in Metridium by prolonged contact with non-clonemates (Purcell, 1977). She recorded changes in size, opacity, and cnidom. Microbasic b-mastigophores and atrichs are arranged from the most mature at the tip to the least at the base (Watson and Mariscal, 1983). The cells migrate to the tip after autonomy so that the catch tentacle becomes effective again. In addition to the observed increase in size and nematocyst content a number of other changes must occur to yield a fully functional catch tentacle. The crossing muscle fibers of Metridium catch tentacle lie very close to the epithelium. The mesoglea is not distinct under the polarizing microscope. It is likely that the connective tissue reduced during development to allow the tentacle great extension and ease of inflation. The tissue was fragile and difficult to manipulate for biomechanical tests. Since the normal tentacles are not affected by Lee: Independent catch tentacle 10 inflation of å catch tentacle, two possible explana¬ tions arise: (1) the increase in pressure to the catch tentacle is very slight, (2) the structure of the feeding tentacle is more rigid such that it requires more pressure than a catch tentacle to extend. Coelenteron pressure has a small pressure range, averaging from 2.6mm to 6.5mm of water (Batham and Pantin, 1950). A better explanation would be that the catch tentacle lost structural support so that small pressure changes can easily affect its size. A catch tentacle can become six times longer in one minute or less. To summarize, a feeding tentacle near the pharynx that is between a pair of perfect mesenteries can be induced to develop into catch tentacle. During this development structural changes in the muscle, the mesoglea, and the nematocysts occur along with the differentiation of a conduction system specific to a catch tentacle. Together a whole complex behavior system emerges that displays action patterns and independent control. Neural recording from the oral disc and tentacles might reveal the properties of the conduction system. Lubbock and Shelton (1981) recorded a triphasic spike in Anthopleura after contact with allogeneic Lee: Independent catch tentacle 11 tissue. Clonemates and objects fail to evoke elec- trical activity. It would be exciting to find the electrical stimulus that will release the agonistic action pattern, beginning with an increase in hydro- static pressure. A closer examination of the histo¬ logical changes in the mesoglea, the musculature, and the nerve net before and after induction will provide insight to the control of catch tentacle activity. I thank Chuck Baxter for his advice on this work and many stimulating discussions, and also for critically reading the manuscript; Freya Sommer for collecting animals; and the people at Hopkins Marine Station for their enthusiasm. Lee: Independent catch tentacle 12 REFERENCES Batham, E.J. and C.F.A. Pantin, 1950. Muscular and hydrostatic action in the sea-anemone Metridium senile (L.). J. exp. Biol. 27:264-289. Batham, E.J. and C.F.A. Pantin, 1951. The organization of the muscular system of Metridium senile. Quart. J. Micro. Sci. 92:27-56. Batham, E.J., et al, 1960. The nerve-net of the sea-anemone Metridium senile: the mesenteries and the column. Quart. J. Micro. Sci. 101:487-510 Francis, L., 1973. Intraspecific aggression and its effects on the distribution of Anthopleura elegantissima and some related sea anemones. Biol. Bull. 144:73-92. Gosse, P.H., 1860. A History of the British Sea Anemones and Corals. Van Voorst, London, 362pp. Hand, C., 1955. Sea anemones of central California coast, part III: The Acontiarian anemones. Wassmann J. of Biol. 13(2):189-251. Lubbock, R. and G.A.B. Shelton, 1981. Electrical activity following cellular recognition of self and non-self in a sea anemone. Nature. 289:59-60. Purcell, J.E., 1977. Aggressive function and induced development of catch tentacles in the sea anemone Metridium senile. Biol. Bull. 153:355-368. Purcell, J.E., 1982. Intraspecific aggression and population distribution of the sea anemone Metridium senile. Biol. Bull. 162:345-359. Robson, E.A., 1971. The behavior and neuromuscular system of Gonactinia prolifera, a swimming sea anemone. J. exp. Biol. 55:611-640. Robson, E.A. and R.K. Josephson, 1969. Neuromuscular properties of mesenteries from the sea-anemone Metridium senile. J. exp. Biol. 50:151-168. C Lee: Independent catch tentacle 13 van Marle, J., 1976. Contribution to the knowledge of the nervous system in the tentacles of some coelenterates. Bijdragen tot de dierkunde. 46(2):219-260. Watson, G.M. and R.N. Mariscal, 1983. Comparative ultrastructure of catch tentacles and feeding tentacles in the sea anemone Haliplanella. Tissue and Cell. 15(6):939-953. Williams, R.B., 1975. Catch tentacles in sea anemones: Occurrence in Haliplanella luciae (Verrill) and a review of current knowledge. 9. Nat. Hist. 9:241-248. Lee: Independent catch tentacle 14 FIGURE CAPTIONS Figure 1 a-i, Behavior sequence of a catch tentacle in aggression. Figure 2 Appearance of catch tentacle, measured from initial contact. Figure 3 Schematic representation of oral disc and distribution of initial catch tentacle response. Figure 1 Musculature of a perfect mesentery, taken from Batham and Pantin, 1951. Figure 5 Longitudinal muscle fibers of regular tentacle. (400 x) Figure 6 Longitudinal and crossing muscle fiber of catch tentacle. (400 x) Lee: Independent catch tentacle 15 TABLE CAI TION Table 1 Response time of Metridium catch tentacle to 10-minute stimulus by Anthopleura. Table 2 Tentacles found between perfect mesenteries. Only the regular tentacles closest to the pharynx were counted. L C ad A Fiodre S 1 S — — 8 ure 2 1 0 + 25 d — — — — Z 1 O — — - N-40 — — POINT OF STIMULUS 66.6% 3.3% 4.8% 4.8% — - — — — IiI 6.6% — — ORAL STOMA RETRACTOR LONGITUDINAL RADIAL PARIETO-BASILAR DISK CIRCULAR Egre Fgre .L Figon 6 — MINUTES 10 — — 2 18 20 25 30 5 NO RESPONSE — o ble NUMBER OF ANEMONES — 2 C — — CATCH TENTACLES REGULAR TENTACLES NUMBER 19/19 20/42 abe O