Jean G. Boal

Cuttlefish use polarization sensitivity in predation on silvery fish

Cephalopods are sensitive to the linear polarization characteristics of light. To examine if this polarization sensitivity plays a role in the predatory behavior of cuttlefish, we examined the preference of Sepia officinalis when presented with fish whose polarization reflection was greatly reduced versus fish whose polarization reflection was not affected. Cuttlefish preyed preferably on fish with normal polarization reflection over fish that did not reflect linearly polarized light (n = 24, chi 2 = 17.3, P < 0.0001), implying that polarization sensitivity is used during predation. We suggest that polarization vision is used to break the countershading camouflage of light-reflecting silvery fish.

The effect of early experience on learning and memory in cuttlefish

The effect of early experience on the growth and ontogeny of memory in cuttlefish (Sepia officinalis) was studied using an associative learning protocol. Five groups of cuttlefish were reared in different conditions (standard conditions, SC; impoverished conditions, IC; enriched conditions, EC; impoverished then enriched conditions, I/EC; enriched then impoverished conditions, E/IC) from birth to the 3rd month of postembryonic life. Acquisition and retention of the learning task were assessed at 1 and 3 months. Growth was slower and maturation of memory abilities occurred later in cuttlefish from Group IC than in cuttlefish from Group EC, with the maturation rate of memory in cuttlefish from Group SC intermediate between these two groups. Retention performances of cuttlefish from Groups I/EC and E/IC indicated that the environment of rearing during the 2nd and/or 3rd months of life was crucial for the development of memory.

Orientation in the cuttlefish Sepia officinalis: response versus place learning

Several studies have demonstrated that mammals, birds and fish use comparable spatial learning strategies. Unfortunately, except in insects, few studies have investigated spatial learning mechanisms in invertebrates. Our study aimed to identify the strategies used by cuttlefish (Sepia officinalis) to solve a spatial task commonly used with vertebrates. A new spatial learning procedure using a T-maze was designed. In this maze, the cuttlefish learned how to enter a dark and sandy compartment. A preliminary test confirmed that individual cuttlefish showed an untrained side-turning preference (preference for turning right or left) in the T-maze. This preference could be reliably detected in a single probe trial. In the following two experiments, each individual was trained to enter the compartment opposite to its side-turning preference. In Experiment 1, distal visual cues were provided around the maze. In Experiment 2, the T-maze was surrounded by curtains and two proximal visual cues were provided above the apparatus. In both experiments, after acquisition, strategies used by cuttlefish to orient in the T-maze were tested by creating a conflict between the formerly rewarded algorithmic behaviour (turn, response learning) and the visual cues identifying the goal (place learning). Most cuttlefish relied on response learning in Experiment 1; the two strategies were used equally often in Experiment 2. In these experiments, the salience of cues provided during the experiment determined whether cuttlefish used response or place learning to solve this spatial task. Our study demonstrates for the first time the presence of multiple spatial strategies in cuttlefish that appear to closely parallel those described in vertebrates.

Experimental evidence for spatial learning in octopuses (Octopus bimaculoides).

Octopuses forage far from temporary home dens to which they return for shelter. Spatial tasks may assess learning. Octopuses (Octopus bimaculoides) were placed in a novel arena, and their movements were tracked for 72 hr. Movements around the arena decreased across time, consistent with exploratory learning. Next, octopuses were given 23 hr to move around an arena; after a 24-hr delay, their memory of a burrow location was tested. Most remembered the location of the open burrow, demonstrating learning in 1 day. Finally, octopuses were trained to locate a single open escape burrow among 6 possible locations. Retention was tested after a week and was immediately followed by reversal training (location rotated 180 degrees ). Octopuses learned the original location of the burrow, remembering it for a week. Path lengths increased significantly after reversal, gradually improving and showing relearning. Octopuses show exploratory behavior, learning, and retention of spatial information.

Behavioral evidence for intraspecific signaling with achromatic and polarized light by cuttlefish (Mollusca: Cephalopoda)

Intraspecific visual communication was studied quantitatively by testing the behavior of Sepia officinalis, the common cuttlefish, as senders and receivers of body pattern signals. These signals can be achromatic and/or linearly polarized and are produced by specialized dermal cells. Experiment 1 addressed whether the presence of conspecifics affected the visible, achromatic body patterns of males. These patterns tended to vary among conditions, suggesting limited sensitivity to audience. One set of body patterns varied with the number of conspecifics viewed while an uncorrelated set of body patterns varied with the sex of conspecifics viewed. Males showed high-contrast zebra banding when viewing another male, supporting previous studies of body patterns in male-male agonistic behavior. Experiment 2 addressed the relationship of polarized light patterns with visible body patterns of males and females, and tested whether senders modified their polarization patterns in response to conspecifics. Polarization patterning was only weakly associated with visible body patterns. Females showed more polarized body patterns than did males, but polarized patterns did not differ among conditions; thus, no sensitivity to audience by senders was found. Experiment 3 addressed whether conspecific receivers used information from polarized body patterns. Limited evidence was found for changes in the behavior of female but not male observers, suggesting that female receivers may use polarized patterns as a source of information about conspecifics. The information contained in polarization patterning may complement that contained in zebra patterning such that both males and females advertise their species, sex, location, and size to conspecific receivers.

Female Choice of Males in Cuttlefish (Mollusca: Cephalopoda)

In captivity, male cuttlefish use visual displays to establish size-based dominance hierarchies and larger males obtain most of the copulations. This experiment was performed to determine if females prefer the larger, more dominant males and whether females use male visual displays to evaluate males. Twenty sexually mature, virgin female cuttlefish were given up to six opportunities to approach one of two males out of a pool of ten adult males, or to approach neither. Females did have significant preferences between males, but they were not related to male dominance. Females showed a consistent and significant preference for the more recently mated male. These preferred males were also more likely to mate again, when given the opportunity, than were less-preferred, less-recently mated males. Females also preferred males that showed fewer zebra displays. Viewing male-male interactions did not affect their choices. Results suggest that female preference could be based on chemical cues, while visual displays may function primarily as agonistic signals. Females who mated did so repeatedly before laying eggs, providing opportunity for sperm competition.

Extreme aggression in male squid induced by a β-MSP-like pheromone.

Male-male aggression is widespread in the animal kingdom and subserves many functions related to the acquisition or retention of resources such as shelter, food, and mates. These functions have been studied widely in the context of sexual selection, yet the proximate mechanisms that trigger or strengthen aggression are not well known for many taxa. Various external sensory cues (visual, audio, chemical) acting alone or in combination stimulate the complex behavioral interactions of fighting behaviors. Here we report the discovery of a 10 kDa protein, termed Loligo β-microseminoprotein (Loligo β-MSP), that immediately and dramatically changes the behavior of male squid from calm swimming and schooling to extreme fighting, even in the absence of females. Females synthesize Loligo β-MSP in their reproductive exocrine glands and embed the protein in the outer tunic of egg capsules, which are deposited on the open sea floor. Males are attracted to the eggs visually, but upon touching them and contacting Loligo β-MSP, they immediately escalate into intense physical fighting with any nearby males. Loligo β-MSP is a distant member of the chordate β-microseminoprotein family found in mammalian reproductive secretions, suggesting that this gene family may have taxonomically widespread roles in sexual competition.

Experimental Evidence for Spatial Learning in Cuttlefish (Sepia officinalis)

Laboratory mazes were used to study spatial-learning capabilities in cuttlefish (Sepia offcinalis), using escape for reinforcement. In preliminary observations, cuttlefish in an artificial pond moved actively around the environment and appeared to learn about features of their environment. In laboratory experiments, cuttlefish exited a simple alley maze more quickly with experience and retained the learned information. Similar improvement was not found in open-field mazes or T mazes, perhaps because of motor problems. Cuttlefish learned to exit a maze that required them to find openings in a vertical wall. The wall maze was modified to an arena, and simultaneous discrimination learning and reversal learning were demonstrated. These experiments indicate that cuttlefish improve performance over serial reversals of a simultaneous, visual-spatial discrimination problem.

Octopuses ( Octopus bimaculoides ) and cuttlefishes ( Sepia pharaonis , S. officinalis ) can conditionally discriminate

In complex navigation using landmarks, an animal must discriminate between potential cues and show context (condition) sensitivity. Such conditional discrimination is considered a form of complex learning and has been associated primarily with vertebrates. We tested the hypothesis that octopuses and cuttlefish are capable of conditional discrimination. Subjects were trained in two maze configurations (the conditions) in which they were required to select one of two particular escape routes within each maze (the discrimination). Conditional discrimination could be demonstrated by selecting the correct escape route in each maze. Six of ten mud-flat octopuses (Octopus bimaculoides), 6 of 13 pharaoh cuttlefish (Sepia pharaonis), and one of four common cuttlefish (S. officinalis) demonstrated conditional discrimination by successfully solving both mazes. These experiments demonstrate that cephalopods are capable of conditional discrimination and extend the limits of invertebrate complex learning.

CONTACT CHEMOSENSORY CUES IN EGG BUNDLES ELICIT MALE-MALE AGONISTIC CONFLICTS IN THE SQUID Loligo pealeii

Male Loligo pealeii engage in frequent agonistic bouts to gain access to female mates while aggregated at communal egg beds. Male squids are attracted to eggs in the field and in the laboratory. It was recently demonstrated that visual detection followed by physical contact with egg capsules elicited male–male aggression. We tested specific physical and chemical features of the egg capsules that may cause this strong behavioral reaction. Male squids were presented with either natural or artificial egg stimuli and scored for four selected behaviors (egg touch, egg blowing, forward-lunge grab, and fin-beating), the last two of which are highly aggressive behaviors. First, squids were presented with natural eggs versus eggs sealed in agarose-coated tubes (ESACT), which eliminated both tactile and chemical stimuli. Second, males were presented with natural eggs versus eggs sealed in agarose coated tubes containing C18 Sep-Pak-purified extracts (TCPE) from squid egg capsules, which provided chemical cues from natural eggs without the physical stimulus of the egg capsules. Third, natural eggs versus heat-denatured eggs were tested to determine whether the active factor in natural eggs is heat-labile. Squids responded aggressively when contacting natural eggs and TCPE, whereas squids did not respond after touching ESACT or denatured eggs. These results suggest that aggressive behavior is elicited by a heat-labile factor that is embedded within squid egg capsules. This chemosensory cue appears to be a contact pheromone that stimulates the agonistic interactions that characterize the mating behavior of migratory squids on inshore spawning grounds.