Vol. 6; Supplement Page 46 THE VELIGER The Light Responses of Tegula funebralis (Mollusca: Gastropoda) DOROTHEA F. KOSIN Hopkins Marine Station of Stanford University, Pacific Grove, California (7 Text figures) night exposure shows great migration downwards during THE BEHAVIOR OF Tegula funebralis (A. ADAMS, 1854) full daylight; the snails gather in the shadiest parts under on the shore is complex. The snails are found clustered algæ, behind rocks, on the shady side of the aquarium. in cracks and crevices of rocks on both sunny and foggy In darkness (with only a dim ruby light for occasional days when the tide is low. Their movements in tidepools observation), the snails all remained at water level all are inconsistent; sometimes all snails are gathered on the the time. Snails kept at a constant low light intensity shady side, at others they are scattered throughout the (about 70 foot candles) tended to stay at the top, though pool. They are subjected to the action of waves and cur- somewhat less than those in complete darkness. rents, as well as tidal exposure and submersion, at dif¬ ferent light conditions over the tidal cycle. To establish irise the role of each factor (such as light) would be difficult in nature, but in the laboratory most of the other factors — can be controlled and light varied. constont dorkness 20 METHODS constant light Tegula funebralis were tested for reactions to light in sev¬ —.—.— eral manners: (1) Three aquaria, as much alike in con¬ tents as possible (rocks, shell fragments, algæe, number of snails) were set up under different light conditions and the snail movements noted. (2) Groups of snails were subjected to different intensities and spectral regions of visible light, and their reactions observed and timed. (3) Individuals were allowed to move in directional light. 310 (4) Eyes and other appendages were removed and the 5 response of such operated snails compared with the nor- doy-night mal. Such operations were not generally harmful: all operated animals survived for at least two weeks and no infections were obvious. (Before operation, the snails were anesthetized by slow addition of an equal volume of isotonic MgCl- solution to sea water in which a number of snails were lying upside down. A temperature of 30° C also served to keep the snails extended out of their shells.) RESULT 12 PM 12M I2PM May May 2 May 3 (1) The experiments with 3 aquaria in different light con- ditions indicate that Tegula funebralis avoid light. Fig. Figure 1: Distribution of 20 Tegula funebralis in each of three shows the number of snails at or near the surface of the similar aquaria under different light conditions: normal day and night; constant illumination at 70 foot candles; and darkness. water (all others being on or near the bottom). The day Vol. 6; Supplement Page 47 THE VELIGER In constant conditions the distribution of Tegula funeb¬ ralis remains constant. When light conditions are alter- 20- nated between light and dark the distribution alternates 50 Foot-candles correspondingly up and down. (Fig. 2) The alternating light was presented out of phase with the day-night cycle and was conducted with snails which had been in the 20 FC. dark for two weeks. The response to the change of light conditions starts immediately, and the new distribution is reached within approximately one hour. 5 Foot-candles (2) Another method for measuring the light response was exposing groups of snails to beams of different inten¬ Note: The following abbreviations are used where appro- priate in all figures in this article. a shell aperture ht head tentacle k kidney an anus cm columellar muscle me mantle cavity et ctenidium op operculum os osphradium e eye darkness foot rectum s shell g gonad o spiral caecum h heart st stomach Time (minutes) Figure 3: Rate of movement of snails out of a beam of light at three light —dark- light —dork- light dork- different intensities (5, 20, 50 f.c.). The rate in darkness is compared. 20 sities of light, and observing their rate of exit from the lighted area. As soon as the snails were placed in a circle of light, they began turning and pushing each other around in their attempt to move out of the light. They move to the darkest areas available, and rest only when they reach a crack or maximum darkness. The same number of snails left in the same area, but without light, disperse much more slowly. (Fig. 3). Snails previously adapted to different light intensities were also tested in this manner (Fig. 4). Those previously exposed to full sunlight (6000 F.C.) scarcely responded at all (3) Both operated and normal snails give a "shadow re- action," which is best observed if the snail is lying on its back, with foot partly extended. (If it is not extended far enough, it will withdraw into the shell when shadowed. If it is extended too far, the snail turns over and will not react at all. When the snail is attached to a surface by its foot, shading causes it to pull back its head, and to remain in that position for some time,— e. g. up to 5 minutes). — But when it is upside down, its normal reaction to a 2M 12PM 12M 12PM 12M 12PM shadow is a slight contraction and pulling back of the foot, followed by a short period of quiet, and then a relax- Figure 2: Effect of alternating light (70 foot candles) and darkness ation or extension of part or all of the foot. There is great on 20 snails that had previously been in darkness for two weeks. The cycle is opposite to that of actual day and night. variation in the amount of contraction, even in the same Page 48 Vol. 6; Supplement THE VELIGER The same procedure was followed at different spectral regions isolated by glass filters; the intensities were ad- 5 Foot-candles 20 Foot-candles justed to give about equal reading on a G. E. light meter, which approximates the human eye over much of 1 the visible range. No significant difference in the duration of the response was noted to shading of either red, yellow, blue or green light (Fig. 6). However, in the far red 5. light from a photographic ruby lamp (10 watts at 1 foot) no shadow response could be elicited. A sudden illumination, even by bright light, does not cause such a consistent and quick reaction; it is the dis¬ appearance of light which causes the withdrawal. Nor does re-illumination visibly affect the total shadowing re¬ Time (minutes) action. If a snail is left in darkness for over an hour, then — darkness ----70 ft. candles ----6000 ft candles put on its back, there may be a long contraction but this cannot be repeated as well as the shadow reaction. During Figure 4: Rate of movement of snails out of a beam of the first minute or longer of such light exposure, the snail light (5 and 20 foot candles), after previous adaptation to may be very insensitive to shadows. The amount of time darkness, 70 foot candles and full daylight (6000 f.c.) snail. The duration also varies, but not necessarily with Epithelium 1o mm the visible degree of contraction. A snail may contract deeply but shortly, or only slightly, but remain still for some time. Fig. 5 indicates the duration of contractions Opening of caused by 19 successive shadowings of 3 different snails; opfic cup Conn the average is 3 to 4 seconds. (The light was a microscope Cornea lamp about 30 cm above the snail, with some 7.5 cm of sea water to absorb much of the infra red. The visible in- tensity at the level of the snail was 20 F.C.) S Figure 7: Sagittal section of the eye lobe of ) Tegula funebralis, about 25 microns thick. necessary for this adaptation is dependent on the bright- ness of the light, and the intensity of light to which the snail had been exposed before the one hour dark period. o Only when the snail becomes adapted to the new intensity Order of shadowing will it react to shadows. (4) In avoiding light, snails move to the closest dark area. Figure 5: Duration of contraction reaction induced by 19 In this they are aided by their eyes. A light directly above successive shadowings of three different snails (represented the snail causes it to move straight out of the light beam in any direction. If the left eye is removed the snail turns by solid, broken and dotted lines). Average duration in about half a circle to the left before moving straight about 3.5 seconds (omitting the four failures, of zero out of range of the light. Similarly a snail turns toward the durations). Light, 20 foot candles. Page 49 Vol. 6; Supplement THE VELIGER Hlongest Fraverage Eshortest Green Blue White Red White - Normal Animals Eyeless Figure 6: The effect of various spectral regions, and of white light, upon the shadowing response. The narrow columns represent different animals (5 to 8 in different histogroms). Eyeless animals are shown in the last group. Intensities approximately equal, by G.E. photo-cell. right when its right eye is removed. When both eyes are There was no difference in their reactions. Figure 2 removed the snail generally turns around irregularly be¬ shows the response of eyeless snails. Apparently the fore leaving the lighted area. shadow reaction is more the result of a general body sen¬ In light from one side a normal snail moves away from sitivity, than of reception by a specific light sensitive organ. the source with its head in its own shadow. The head turns These experiments indicate that the eyes are used for from side to side, and each time that an eye is illuminated directional orientation in the light. It is the only function the snail turns back to its shadow or turns its head to the of the eye that I have been able to discover. The anatomy other side. In the same light an eyeless snail moves of the eye suggests this as well (Fig. 7). It is a simple. irregularly, even directly toward the light. Apparently it small ocellus containing a firm, clear substance which may cannot distinguish direction without its eyes. A snail with- correspond to a “lens". The small opening to the outside out its right eye will turn, usually toward the right, until might function like a pin-hole camera, to form an image its left eye is in shadow and then move directly out of at the retina. Around the edge of the opening is a ring the path of the light. When the left eye is removed the of unpigmented cells which might be a cornea. The re¬ snail turns, usually to the left, until its right eye is in tina appears to be formed by a direct invagination of the shadow before moving straight out of the beam of light. epidermis, continuous through the clear corneal area Normal snails, snails without eyes, without epipodial The optic nerve branches proximal to the optic cup, tentacles and lobes, without propodial tubercles, and with sending fibers over its entire surface to connect with the none of these organs were tested for a shadow reaction. retina. Page 50 Vol. 6; Supplement THE VELIGER The eyes of Tegula funebralis may represent a concen¬ SUMMARY tration of the same sensitive tissue present on the general body normally outside of the shell. The pigmentation of Tegula funebralis is negatively phototactic. It becomes the retina resembles that of the epidermis, which contains adapted to bright light, and loses its sensitivity temporar¬ melanin. Attempts to isolate a photosensitive pigment ily. It has a general body sensitivity shown by a shadow from the eyes, according to the recipe for squid retinas reaction. Its eyes, simple lens ocelli, seem responsible for (BLISS, 1948) failed, probably because of the very small directional orientation to light. Without eyes its orienta- amount of tissue employed. tion is irregular. LITERATURE CITED BLISS, A. F. 1948. The absorption products of visual purple of the squid and its bleaching products. Journ. Biol. Chem. 176: 563 - 569 Observations on the Epipodium, Digestive Tract, Coelomic Derivatives and Nervous System of the Trochid Gastropod Tegula funebralis JOHN A. MACDONALD AND C. BURKE MAINO Hopkins Marine Station of Stanford University, Pacific Grove, California (15 Text figures) to determine the extent of specific body cavities. Both IN SPITE of the abundance of Tegula funebralis (A. Mallory’s connective tissue stain and Harris' hematoxylin ADAMS, 1854) in the California intertidal, no account of its produced excellent results after fixation with Bouin’s fluid, anatomy has yet appeared in the literature, although the made with seawater. A silver impregnation (RowELL, anatomy of other trochids has been described (Randles, 1963) worked well for isolated nerves, but also stained 1905; Fretter and Graham, 1962). To partially fill this muscle and connective tissue heavily in sections of the void, a brief account of certain external and internal fea¬ entire animal tures of T. funebralis is herein presented Epipodium: The epipodium of Tegula funebralis is com¬ The specimens of Tegula funebralis examined were col¬ posed of five elements: the neck lobes, anterior papillae, lected at Mussel Point, Pacific Grove, California, during April and May, 1963. The animals were dissected alive epipodial tentacles, epipodial papillae, and epipodial ridges (Figures 1 and 2). On both sides of the animal after having been anesthetized with magnesium chloride the anterior quarter of the epipodium is occupied by both frozen and paraffin sections were cut in order to the heavily ciliated neck lobe, which runs posteriorly make more detailed observations. Injection of suspension from near the base of the optic peduncle. The border of of carborundum and carmine powders in sea water helped Page 51 Vol. 6; Supplement THE VELIGER these neck lobes may be the removal of particles expelled from the mantle cavity, though FRETTER & GRAHAM DG HG (1962, p. 532) state that the neck lobes of British trochids are rolled into half-siphons for channeling water in and out of the mantle cavity. No evidence of this was seen in ME Tegula funebralis. Beneath the overhang of each neck lobe are from one CL to ten anterior papillae, each with a central spot of ciliated unpigmented epithelium (Fig. 3). They retract slightly Bvgan CM when touched; this reaction and their structure suggest — that they may be sensory receptors. HT NL ET PE Figure 1: Entire animal, left side, shell removed QUE G gonad C ctenidium CT HG hypobranchial gland CL cephalic lappet HT cephalic tentacle CM columellar muscle DG digestive gland M mouth ME mantle edge E eye NL neck lobe ER epipodial ridge OP operculum ET epipodial tentacle SC spiral caecum Figure 3: Anterior papilla, diagrammatic longitudinal section the left neck lobe is fringed, the number and grouping CUE ciliated unpigmented CT connective tissue of points being variable, whereas the edge of the right lobe epithelium PE pigmented epithelium is smooth; on both lobes the cilia beat distally, especially when the lobe is touched with a probe. The function of Just posterior to the neck lobe is the first epipodial tentacle, which on the left side bears an epipodial papilla (Fig. 6) similar in appearance to an anterior papilla; DG - HG the first epipodial tentacle on the right side does not bear such a papilla. The other three tentacles on each side bear papillae (Fig. 4). The epipodial tentacles are similar in structure to the cephalic tentacles and are innervated from the pedal cords; observations of the use of both ME cephalic and epipodial tentacles in the field and in aqua¬ - CL CM ER HT - ET NL E ER Figure 2: Entire animal, right side, shell removed. e HT cephalic tentacle CL cephalic lappet LK left kidney CM columellar muscle M mouth E eye ME mantle edge ER epipodial ridge Figure 4: Epipodial tentacle, diagrammatic NL neck lobe ET epipodial tentacle ET epipodial tentacle EP epipodial papilla OP operculum G gonad SP sensory papillae ER epipodial ridge HG hypobranchial gland R rectum Page 52 Vol. 6; Supplement THE VELIGER ria seem to indicate that they are tactile, and perhaps olfactory, receptors. An active animal will be seen to draw its tentacles repeatedly over the substrate as it advances. Resting animals either “caress“ their shells with the ten- tacles, or gently wave them. In the presence of predaceous asteroids such as Pisaster ochraceus (BRANDT, 1835), T. funebralis waves cephalic and epipodial tentacles vigor¬ ously while trying to escape; Mc GEE (unpubl.) has noted a similar response in Tegula brunnea (PHILIPPI, 1848) ex- OS- posed to spawning males of their own species. — HG LORK LM -OLK C -PE RR LK EBV PC MC SC V DG Figure 5: Epipodial tentacle, diagrammatic cross section H haemocoel MC column of transverse LM longitudinal muscle muscle and connective band tissue N nerve PE pigmented epithelium Figure 7: Dorsal view of entire animal with mantle cavity laid open; structures of coelomic origin are indicated C ctenidium OLK left kidney opening DG digestive gland ORK right kidney opening EBV efferent branchial OS osphradium PE¬ vein PC pericardium F foot R rectum G gonad RK right kidney HG hypobranchial gland RS radular sac LK left kidney SC spiral caecum The epithelium of the epipodial tentacles is heavily pig¬ mented; distally it forms papillae bearing non-motile sen¬ sory cilia. Similar cilia are found on the tentacles of other prosobranchs (FRETTER & GRAHAM, 1962, p. 313) and on the oral tentacles of the nudibranch Hermissenda (AGERS- BORG, 1925). Passing down the length of the tentacle is à large central nerve, longitudinal muscle bands, and an Figure 6: Epipodial papilla, diagrammatic extension of the hemocoel. The center of the tentacle is longitudinal section occupied by an irregular column of muscle and con¬ CT connective tissue PE pigmented epithelium nective tissue fibers (Fig. 5). The epipodial ridge is a N nerve UE unpigmented epithelium long flap of tissue which begins just posterior to the first Page 53 Vol. 6; Supplement THE VELIGER epipodial tentacle on either side and runs just dorsal to the other three epipodial tentacles and continues to the hindmost tip of the foot. 7 -BW Digestive Tract: The anterior part of the digestive system seen in Figure 8, shows the buccal cavity to be limited — ME ventrally by the subradular membrane, which extends HARC RA¬ posteriorly into the radular sac. Ventrally, the radular sac is anchored by striated musculature to the posterior part — SG N DSG of the odontophore. The radula is fused to the subradular membrane anteriorly, but is free posteriorly. The com¬ ARC plex movements of the odontophore are controlled by striated musculature acting on the four radular "carti¬ lages. The disposition of the rest of the digestive system is seen in outline in Figure 9. There are three main regions: the foregut, composed of buccal cavity and esophagus; the stomach and digestive gland, which comprise the mid¬ DG RA ES ST- C — Figure 9: Dorsal view of entire digestive system ARC anterior radular carti- I intestine lage M mouth BW cut edge of body wall ME mantle edge 2 . . .. DG digestive gland RA radula 2 DSG duct of salivary gland SC spiral caecum G gonad SG salivary gland R rectum ST stomach PRC BC SRM ARC Figure 8: Buccal region, diagrammatic PE longitudinal section Vgggvvote ARC anterior radular carti-M mouth NNNN000 lage PRC posterior radular BC buccal cavity cartilage ES esophagus RA radula Figure 10: Cross section through esophagus J jaw RS radular sac SRM subradular membrane gut region; and the hindgut, consisting of intestine and rectum. Only one duct is shown leading to the digestive gland from the stomach; there are also numerous fine 1 pores in the same area which appear to lead to the gland. The areas of the posterior visceral hump occupied by the digestive gland and gonad vary somewhat from spec- ST imen to specimen. In freshly killed specimens which were not treated with magnesium chloride, peristaltic move¬ Figure 11: Cross section through spiral caecum ments could be seen in the hindgut, but were not observed in the foregut. SC spiral caecum ST stomach Page 54 THE VELIGER Vol. 6; Supplement ER -CA H ETA ar V MA EP¬ M CC PE- Figure 12: Cross section through rectum PN¬ er CA cilia CT connective tissue RK LK M Figure 14: Left side of foot, diagrammatic cross section LRPI CC cross connection bet¬ ET epipodial tentacle ween pedal cords H haemocoel EBV CT connective tissue MA muscular area ZRRp. EN epipodial nerve P pedal cord EP epipodial papilla PE pigmented epithelium ER epipodial ridge PN pedal nerve I- relatively large, and fluid may be caused to flow between the left kidney and the pericardial space. Nervous System: The nervous system of Tegula funebra¬ RK lis (Figs. 14 & 15) is similar to that of other trochids AO (RANDLES, 1905, pp. 57-66, figs. 30-33). The right pleuro¬ parietal connective runs over the esophagus and splits Figure 13: Diagrammatic dorsal view of pericardia into two nerves, the first running to the branchial gan¬ glion, subjacent to the osphradium, and the second run¬ cavity showing coelomic derivatives ning posteriorly between the ventral ctenidial membrane A auricle PC pericardium and the perivisceral sinus, and crossing the esophagus AO aorta RK right kidney and the loop of the hindgut to end in a pair of visceral EBV efferent branchial RRPP right renopericardial ganglia above the right kidney. The left pleuro-visceral vein pore connective crosses beneath the esophagus and runs poste¬ I intestine LRPP left renopericardial riorly to connect with the visceral ganglia. RANDLES LK left kidney pore (1905) found a dialyneury between the left pallial nerve Vventricle and the right pleuro-parietal connective in Trochus; no such connection was observed in Tegula funebralis, Coelomic Derivatives: Structures of cœlomic origin are shown in figure 13; these include the right and left kid¬ ACKNOWLEDGMENTS neys, the pericardial cavity, and the gonad. The presence We are indebted to the faculty, staff, and graduate stu¬ of both right and left renopericardial ducts is in accord with the condition found in other trochids (RANDLES. dents of the Hopkins Marine Station, especially to Mr. 1905) and in Haliotis (HARRISON, 1961). The duct to Nick Holland and Mr. Welton Lee, for advice, technical the right kidney is very small; whether it is functional or assistance, and encouragement in the execution of this work. not is undetermined. The opening to the left kidney is Vol. 6; Supplement LN CB -LC CG APN LPN PRPN CPL RPP BRG LPV EN¬ PN¬ Figure 15: Nervous system, diagrammatic dorsal view. The large masses dorsal to the pedal ganglia represent the pleural ganglia; the small hollow objects between the pleural ganglia are the statocysts APN anterior pedal nerve LN labial nerve BG buccal ganglion LPN left pallial nerve BRG branchial ganglion LPV left pleuro-visceral CB cerebral commissure connective CC cross connection bet¬ ON optic nerve ween pedal cords P pedal cord CG cerebral ganglion PG pedal ganglion CP cerebro-pedal connect- PN pedal nerve RPN right pallial nerve CPL cerebro-pleural con¬ RPP right pleuro-parietal nective connective EN epipodial nerve TN tentacle nerve LC labial commissure VG visceral ganglia Page 55 THE VELIGER LITERATURE CITED AGERSBORG, H. P KJERSCHOW 1925. The sensory receptors and the structure of the oral tentacles of the nudibranchiate mollusk Hermissenda crassi- cornis (EsCHSCHOLTz, 1831). Acta Zool. 6: 167 - 182 FRETTER, VERA, & ALASTAIR GRAHAM 1962. British prosobranch molluscs; their functional anatomy and ecology. London, Ray. Soc.; xvi + 755 pp.; 317 figs. HARRISON, F M. 1961. Some excretory processes in the abalone Haliotis ruf¬ escens. Journ. Exp. Biol. 39: 179-192 RANDLES, W. B. 1905. Some observations on the anatomy and affinities of the Trochidae. Quart. Journ. micr. Sci. 48: 33 -78 ROWELL, C. H. F. 1963. A new technique for silvering invertebrate central ner¬ vous systems. Quart. Journ. micr. Sci. 104: 81 - 87