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Octopus arms, notable for their complex anatomy and remarkable flexibility, have sparked significant interest within the neuroscience community. However, there remains a dearth of knowledge about the neurochemical organization of various cell types in the arm's nervous system. To address this gap, we used hybridization chain reaction (HCR) to identify distinct neuronal types in the axial nerve cords of the pygmy octopus, Octopus bocki, including putative dopaminergic, octopaminergic, serotonergic, GABAergic, glutamatergic, cholinergic, and peptidergic cells. We obtained high-resolution multiplexed fluorescent images at 0.28 × 0.28 × 1.0 μm voxel size from 10 arm base and arm tip cross sections (each 50 μm thick) and created three-dimensional reconstructions of the axial ganglia, illustrating the spatial distribution of multiple neuronal populations. Our analysis unveiled anatomically distinct and molecularly diverse scattered neurons, while also highlighting multiple populations of dense small neurons that appear uniformly distributed throughout the cortical layer and potential glial cells in the neuropil. Our data provide new insights into how different types of neurons may contribute to an octopus's ability to interact with its environment and execute complex tasks. In addition, our findings establish a benchmark for future studies, allowing pioneering exploration of octopus arm molecular neuroanatomy and offering exciting new avenues in invertebrate neuroscience research., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

The peripheral nervous system of the octopus is among the most complex of any animal. In each arm, hundreds of serial ganglia form a central core of nervous tissue processing sensory input, issuing motor commands, and exchanging information with the central brain. 1 , 2 , 3 , 4 , 5 In addition to the central cord, there are two other types of neural elements: fine intramuscular nerve cords (INCs) 6 , 7 and small sucker ganglia at the base of each sucker. 2 , 6 , 8 , 9 Connections between these different elements and the structural organization of the arm nervous system remain poorly understood, despite decades of interest and a more recent explosion of studies of the cephalopod nervous system. 8 , 10 , 11 , 12 , 13 , 14 , 15 Here, we use serial blockface electron microscopy to reconstruct large volumes of an arm from Octopus bocki at the base and toward the tip, mapping connections between the various neural elements and their relationship to the muscle and skin. We show that the ganglia follow an alternating mirror-image pattern along the arm, where the left or right-sided location of successive suckers determines ganglionic orientation. We also describe previously unrecognized patterns in (1) continuity of oblique connectives between the INCs that encircle the arm; (2) repeatable structures of the major blood vessel branches and nerve connectives within each ganglion; (3) clustering of rare, unusually large neurons within the cell body layers; and (4) division of the cortex into repeating columns. These new findings from the first 3DEM reconstruction of the arm should greatly facilitate future studies of octopus neurobiology, particularly sensori-motor integration and arm control., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Muscular hydrostats, such as octopus arms or elephant trunks, lack bones entirely, endowing them with exceptional dexterity and reconfigurability. Key to their unmatched ability to control nearly infinite degrees of freedom is the architecture into which muscle fibers are weaved. Their arrangement is, effectively, the instantiation of a sophisticated mechanical program that mediates, and likely facilitates, the control and realization of complex, dynamic morphological reconfigurations. Here, by combining medical imaging, biomechanical data, live behavioral experiments, and numerical simulations, an octopus-inspired arm made of [Formula: see text]200 continuous muscle groups is synthesized, exposing "mechanically intelligent" design and control principles broadly pertinent to dynamics and robotics. Such principles are mathematically understood in terms of storage, transport, and conversion of topological quantities, effected into complex 3D motions via simple muscle activation templates. These are in turn composed into higher-level control strategies that, compounded by the arm's compliance, are demonstrated across challenging manipulation tasks, revealing surprising simplicity and robustness., Competing Interests: Competing interests statement:The authors declare no competing interest.

The horned octopod Eledone cirrhosa, a medium-sized species found in Arabian Gulf off Saudi Arabia, was collected monthly from the Arabian Gulf off Dammam city during January to December 2022. Samples were dissected and prepared for examination using transmission electron microscopy. During genital maturation, seminiferous tubules are established in the testis, with active spermatogonia dividing. Spermatocytes 1 are observed in the tubule, followed by an increase in spermatogonia and spermatocytes in August. Spermiogenesis begins, with spherical spermatids evolving into elongated spermatids. In September, active spermatogonia, meiotic divisions, and increased spermiogenesis continue. Spermatozoa appear in Needham's pouch, indicating sexual maturity. The ovary undergoes various stages of development, with oocytes at stage I in June and July, followed by stage II in October and November. In stage III, follicular cords invade the oocyte's cytoplasm, forming numerous lipid inclusions and protein granules. The cytoplasm contains cisternae of endoplasmic reticulum and a poorly developed Golgi apparatus. Stage IV occurs in November, characterized by the maximum development of follicular cords and the beginning of vitellogenesis. The ooplasm contains numerous lipid inclusions, a syncytium, and secretory cells. From December, stage V oocytes are mainly present, indicating the activity phase of maximum secretion. Yolk platelets accumulate in the oocyte ooplasm, and chorion forms at the zona pellucida. In January, the first smooth eggs are found in some octopuses' ovary, with their proportion increasing steadily. This study aimed to investigate the mitogenic action of gonadotropin and identify the periods of intense cell multiplication during the sexual cycle using cytological methods.

Bioinspired and biomimetic soft grippers are rapidly growing fields. They represent an advancement in soft robotics as they emulate the adaptability and flexibility of biological end effectors. A prominent example of a gripping mechanism found in nature is the octopus tentacle, enabling the animal to attach to rough and irregular surfaces. Inspired by the structure and morphology of the tentacles, this study introduces a novel design, fabrication, and characterization method of dielectric elastomer suction cups. To grasp objects, the developed suction cups perform out-of-plane deflections as the suction mechanism. Their attachment mechanism resembles that of their biological counterparts, as they do not require a pre-stretch over a rigid frame or any external hydraulic or pneumatic support to form and hold the dome structure of the suction cups. The realized artificial suction cups demonstrate the capability of generating a negative pressure up to 1.3 kPa in air and grasping and lifting objects with a maximum 58 g weight under an actuation voltage of 6 kV. They also have sensing capabilities to determine whether the grasping was successful without the need of lifting the objects., (Creative Commons Attribution license.)

Coleoid cephalopods have a high intelligence, complex structures, and large brain. The cephalopod brain is divided into supraesophageal mass, subesophageal mass and optic lobe. Although much is known about the structural organization and connections of various lobes of octopus brain, there are few studies on the brain of cephalopod at the molecular level. In this study, we demonstrated the structure of an adult Octopus minor brain by histomorphological analyses. Through visualization of neuronal and proliferation markers, we found that adult neurogenesis occurred in the vL and posterior svL. We also obtained specific 1015 genes by transcriptome of O. minor brain and selected OLFM3, NPY, GnRH, and GDF8 genes. The expression of genes in the central brain showed the possibility of using NPY and GDF8 as molecular marker of compartmentation in the central brain. This study will provide useful information for establishing a molecular atlas of cephalopod brain., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Octopods are masters of camouflage and solve complex tasks, and their cognitive ability is said to approach that of some small mammals. Despite intense interest and some research progress, much of our knowledge of octopus neuroanatomy and its links to behavior and ecology comes from one coastal species, the European common octopus, Octopus vulgaris. Octopod species are found in habitats including complex coral reefs and the relatively featureless mid-water. There they encounter different selection pressures, may be nocturnal or diurnal, and are mostly solitary or partially social. How these different ecologies and behavioral differences influence the octopus central nervous system (CNS) remains largely unknown. Here we present a phylogenetically informed comparison between diurnal and nocturnal coastal and a deep-sea species using brain imaging techniques. This study shows that characteristic neuroanatomical changes are linked to their habits and habitats. Enlargement and division of the optic lobe as well as structural foldings and complexity in the underlying CNS are linked to behavioral adaptation (diurnal versus nocturnal; social versus solitary) and ecological niche (reef versus deep sea), but phylogeny may play a part also. The difference between solitary and social life is mirrored within the brain including the formation of multiple compartments (gyri) in the vertical lobe, which is likened to the vertebrate cortex. These findings continue the case for convergence between cephalopod and vertebrate brain structure and function. Notably, within the current push toward comparisons of cognitive abilities, often with unashamed anthropomorphism at their root, these findings provide a firm grounding from which to work., Competing Interests: Declaration of interests The authors declare no competing interests., (Crown Copyright © 2021. Published by Elsevier Inc. All rights reserved.)

Cephalopods have evolved nervous systems that parallel the complexity of mammalian brains in terms of neuronal numbers and richness in behavioral output. How the cephalopod brain develops has only been described at the morphological level, and it remains unclear where the progenitor cells are located and what molecular factors drive neurogenesis. Using histological techniques, we located dividing cells, neural progenitors and postmitotic neurons in Octopus vulgaris embryos. Our results indicate that an important pool of progenitors, expressing the conserved bHLH transcription factors achaete-scute or neurogenin , is located outside the central brain cords in the lateral lips adjacent to the eyes, suggesting that newly formed neurons migrate into the cords. Lineage-tracing experiments then showed that progenitors, depending on their location in the lateral lips, generate neurons for the different lobes, similar to the squid Doryteuthis pealeii . The finding that octopus newborn neurons migrate over long distances is reminiscent of vertebrate neurogenesis and suggests it might be a fundamental strategy for large brain development., Competing Interests: AD, RS, AE, GM, ES No competing interests declared, (© 2021, Deryckere et al.)

Octopuses have eight radially symmetrical arms that surround the base of a bilaterally symmetrical body. These numerous appendages, which explore the environment, handle food, and defend the animal against predators, are highly susceptible to truncation or loss. Here, we used scaling relationships specific to the arms of three sympatric octopus species of the genus Octopus, to calculate the proportion of arm truncation. We then compared the frequency and proportion of arm losses between different body locations. Truncated arms were found in 59.8 % of specimens examined, with individuals bearing one to as many as seven injured arms. We found a significant left side bias for greater proportion of arm truncation for all species and sexes except in O. bimaculatus males. We also found that sister species O. bimaculatus and O. bimaculoides had a greater proportion of their anterior arms (pairs 1 and 2) truncated, while in O. rubescens, posterior arms (pairs 3 and 4) were more truncated. The mean percent of arm that was truncated was 28.1 % overall but varied between species and by sex and was highest in O. rubescens females (56 %). The arms of O. rubescens also exhibited the steepest scaling patterns, and showed a positive correlation between body size and number of truncated arms. Overall, we show that arm injuries in our sampling of three intertidal species are frequent and asymmetrical, and that when injured, octopus on average lose a considerable proportion of their arm. Through quantifying the variation in arm truncation, this study provides a new foundation to explore behavioral compensation for arm loss in cephalopods., (Copyright © 2021 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Purpose: Review octopus ocular anatomy and describe the histopathologic findings in three octopuses diagnosed with phakitis and retinitis., Animals: Two common octopuses (Octopus vulgaris) and one giant Pacific octopus (Enteroctopus dofleini) with a history of ophthalmic disease., Methods: A literature search was performed for the ocular anatomy section. Both eyes from all three octopuses, and two control eyes, were submitted for histopathologic evaluation. Hematoxylin and eosin stain was used for standard histopathologic evaluation; GMS stain was used to screen for fungi, gram stain for bacteria; and Fite's acid fast stain for acid fast bacteria., Results: Anatomically, the anterior chamber of the octopus has direct contact with ambient water due to an opening in the dorsal aspect of a pseudocornea. The octopus lens is divided into anterior and posterior segments. The anterior half is exposed to the environment through the opening into the anterior chamber. Neither part of the lens has a lens capsule. The retina is everted, unlike the inverted vertebrate retina, and consists of just two layers. Histopathology revealed inflammatory phakitis and retinitis of varying severity in all six eyes of the study animals. No intraocular infectious organisms were recognized but one common octopus eye had clusters of coccidian parasites, identified as Aggregata sp., in extraocular tissues and blood vessels., Conclusion: We describe inflammatory phakitis and retinitis in two species of octopuses. The underlying cause for the severe intraocular response may be direct intraocular infection, water quality, an ocular manifestation of a systemic disease, or natural senescence., (© 2021 American College of Veterinary Ophthalmologists.)

Octopuses are active predators with highly flexible bodies and rich behavioral repertoires [1-3]. They display advanced cognitive abilities, and the size of their large nervous system rivals that of many mammals. However, only one third of the neurons constitute the CNS, while the rest are located in an elaborate PNS, including eight arms, each containing myriad sensory receptors of various modalities [2-4]. This led early workers to question the extent to which the CNS is privy to non-visual sensory input from the periphery and to suggest that it has limited capacity to finely control arm movement [3-5]. This conclusion seemed reasonable considering the size of the PNS and the results of early behavioral tests [3, 6-8]. We recently demonstrated that octopuses use visual information to control goal-directed complex single arm movements [9]. However, that study did not establish whether animals use information from the arm itself [9-12]. We here report on development of two-choice, single-arm mazes that test the ability of octopuses to perform operant learning tasks that mimic normal tactile exploration behavior and require the non-peripheral neural circuitry to use focal sensory information originating in single arms [1, 10]. We show that the CNS of the octopus uses peripheral information about arm motion as well as tactile input to accomplish learning tasks that entail directed control of movement. We conclude that although octopus arms have a great capacity to act independently, they are also subject to central control, allowing well-organized, purposeful behavior of the organism as a whole., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Animals display wide-ranging evolutionary adaptations based on their ecological niche. Octopuses explore the seafloor with their flexible arms using a specialized "taste by touch" system to locally sense and respond to prey-derived chemicals and movement. How the peripherally distributed octopus nervous system mediates relatively autonomous arm behavior is unknown. Here, we report that octopus arms use a family of cephalopod-specific chemotactile receptors (CRs) to detect poorly soluble natural products, thereby defining a form of contact-dependent, aquatic chemosensation. CRs form discrete ion channel complexes that mediate the detection of diverse stimuli and transduction of specific ionic signals. Furthermore, distinct chemo- and mechanosensory cells exhibit specific receptor expression and electrical activities to support peripheral information coding and complex chemotactile behaviors. These findings demonstrate that the peripherally distributed octopus nervous system is a key site for signal processing and highlight how molecular and anatomical features synergistically evolve to suit an animal's environmental context., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

In medical robotics, micromanipulation becomes particularly challenging in the presence of blood and secretions. Nature offers many examples of adhesion strategies, which can be divided into two macro-categories: morphological adjustments and chemical adaptations. This paper analyzes how two successful specializations from different marine animals can converge into a single biomedical device usable in moist environments. Taking inspiration from the morphology of the octopus sucker and the chemistry of mussel secretions, we developed a protein-coated octopus-inspired micro-sucker device that retains in moist conditions about half of the adhesion it shows in dry environments. From a robotic perspective, this study emphasizes the advantages of taking inspiration from specialized natural solutions to optimize standard robotic designs.

The development of the epidermis of octopus, Octopus vulgaris, throughout its life cycle was studied by conventional staining and histochemical techniques using lectins. The mantle, the arm and the two parts of the suckers: the infundibulum and the acetabulum were analysed independently. With the exception of the suckers, the general morphology of the epidermis does not vary from the first days post-hatching to adulthood. In general terms, histochemical techniques do not indicate changes in the composition of glycoconjugates of the epidermis main cells, epithelial and secretory cells. The epithelial cells of the mantle and arm show positivity for mannose (ConA+) in their apical portions, indicating the presence of n-glycoproteins that, among other things, provide lubrication to the surface of the body. In the suckers, the apical surface of the infundibulum contains sulphated glycosaminoglycans of the N-acetylglucosamine type that provide adhesive properties. In addition to observing three types of mucocytes, m1 and m2 are characteristic of the mantle and arm, and m3 is found in the suckers. The paralarva epidermis is characterised by the presence of Kölliker's organs whose exact function is unknown. In this study, the absence of staining with alcian blue/periodic acid-Schiff(AB/PAS) prevents the possibility of attributing a secretory function. Nevertheless, the linkage of three lectins (WGA, LEL and GSL-I) in the fascicle of the organ suggests the presence of proteoglycans rich in N-acetylglucosamine that would mainly have a structural role., (© 2020 Blackwell Verlag GmbH.)

The vacuum chuck is widely used in industrial and daily life. By observing the macroscopic and microscopic morphology of octopus sucker, it is found that the sucker surface has concave-convex continuous wave shape with large number of non-smooth morphologies. The sealing mechanism of octopus sucker is analyzed according to its surface morphology before and after adsorption, and the non-smooth morphology is found to greatly enhance the adsorption. Based on the bionics theory, the non-smooth surface morphology of octopus sucker is applied to improve the sucker adsorption. And the bionic suckers with three types of grooves are designed. According to the model of standard and bionic suckers, the sucker entities are obtained by the method of three-dimensional printing and casting. And the tensile tests of suckers are carried out. The stress of suckers is analyzed by finite element method, and the sealing mechanism is discussed. According to the test results, the bionic sucker has larger adsorption force. And the ring sucker possesses the best adsorption performance. Compared with the standard sucker, the maximum adsorption force of the bionic sucker is increased by 12.2% in the air and 25.2% underwater. The adsorption force of bionic sucker becomes larger with the increase in the groove number; when the groove number increases to a certain extent, the adsorption force becomes smaller. The deformation of non-smooth morphology during adsorption makes the bionic sucker have a larger contact area. That is the reason why the bionic sucker has good adsorption performance. The bionic design of sucker can provide a new method to improve its adsorption.

Spontaneous nerve regeneration in cephalopod molluscs occurs in a relative short time after injury, achieving functional recovery of lost capacity. In particular, transection of the pallial nerve in the common octopus ( Octopus vulgaris ) determines the loss and subsequent restoration of two functions fundamental for survival, i.e. breathing and skin patterning, the latter involved in communication between animals and concealment. The phenomena occurring after lesion have been investigated in a series of previous studies, but a complete analysis of the changes taking place at the level of the axons and the effects on the animals' appearance during the whole regenerative process is still missing. Our goal was to determine the course of events following injury, from impairment to full recovery. Through imaging of the traced damaged nerves, we were able to characterize the pathways followed by fibres during regeneration and end-target re-innervation, while electrophysiology and behavioural observations highlighted the regaining of functional connections between the central brain and periphery, using the contralateral nerve in the same animal as an internal control. The final architecture of a fully regenerated pallial nerve does not exactly mirror the original structure; however, functionality returns to match the phenotype of an intact octopus with no observable impact on the behaviour of the animal. Our findings provide new important scenarios for the study of regeneration in cephalopods and highlight the octopus pallial nerve as a valuable 'model' among invertebrates., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Multiple sclerosis (MS) is an immune-mediated disease which can cause different symptoms due to the involvement of different regions of the central nervous system (CNS). Although this disease is characterized by the demyelination process, the most important feature of the disease is its degenerative nature. This nature is clinically manifested as progressive symptoms, especially in patients' walking, which can even lead to complete debilitation. Therefore, finding a treatment to prevent the degenerative processes is one of the most important goals in MS studies. To better understand the process and the effect of drugs, scientists use animal models which mostly consisting of mouse, rat, and monkey. In evolutionary terms, octopuses belong to the invertebrates which have many substantial differences with vertebrates. One of these differences is related to the nervous system of these organisms, which is divided into central and peripheral parts. The difference lies in the fact that the main volume of this system expands in the limbs of these organisms instead of their brain. This offers a kind of freedom of action and processing strength in the octopus limbs. Also, the brain of these organisms follows a non-somatotopic model. Although the complex actions of this organism are stimulated by the brain, in contrast to the human brain, this activity is not related to a specific region of the brain; rather the entire brain area of the octopus is activated during a process. Indeed, the brain mapping or the topological perception of a particular action, such as moving the limbs, reflects itself in how that activity is distributed in the octopus brain neurons. Accordingly, various actions are known with varying degrees of activity of neurons in the brain of octopus. Another important feature of octopuses is their ability to regenerate defective tissues including the central and peripheral nervous system. These characteristics raise the question of what features can an octopus show when it is used as an organism to create experimental autoimmune encephalomyelitis (EAE). Can the immune system damage of the octopus brain cause a regeneration process? Will the autonomy of the organs reduce the severity of the symptoms? This article seeks to provide evidence to prove that use of octopuses as laboratory samples for generation of EAE may open up new approaches for researchers to better approach MS., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Here, we report the findings after application of neurobiotin tracing to pallial and stellar nerves in the mantle of the cephalopod mollusk Octopus vulgaris and to the axial nerve cord in its arm. Neurobiotin backfilling is a known technique in other molluscs, but it is applied to octopus for the first time to be best of our knowledge. Different neural tracing techniques have been carried out in cephalopods to study the intricate neural connectivity of their nervous system, but mapping the nervous connections in this taxon is still incomplete, mainly due to the absence of a reliable tracing method allowing whole-mount imaging. In our experiments, neurobiotin backfilling allowed: (1) imaging of large/thick samples (larger than 2 mm) through optical clearing; (2) additional application of immunohistochemistry on the backfilled tissues, allowing identification of neural structures by coupling of a specific antibody. This work opens a series of future studies aimed to the identification of the neural diagram and connectome of octopus nervous system.

Octopus and cephalopods are able to regenerate injured tissues. Recent advancements in the study of regeneration in cephalopods appear promising encompassing different approaches helping to decipher cellular and molecular machinery involved in the process. However, lack of specific markers to investigate degenerative/regenerative phenomena and inflammatory events occurring after damage is limiting these studies. Label-free multiphoton microscopy is applied for the first time to the transected pallial nerve of Octopus vulgaris Various optical contrast methods including coherent anti-Stokes Raman scattering (CARS), endogenous two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) have been used. We detected cells and structures often not revealed with classical staining methods. CARS highlighted the involvement of haemocytes in building up scar tissue; CARS and TPEF facilitated the identification of degenerating fibres; SHG allowed visualization of fibrillary collagen, revealing the formation of a connective tissue bridge between the nerve stumps, likely involved in axon guidance. Using label-free multiphoton microscopy, we studied the regenerative events in octopus without using any other labelling techniques. These imaging methods provided extremely helpful morpho-chemical information to describe regeneration events. The techniques applied here are species-specific independent and should facilitate the comparison among various animal species., (© 2018 The Author(s).)

Octopus vulgaris underwent a radical modification to cope with the benthic lifestyle. It diverged from other cephalopods in terms of body plan, anatomy, behavior, and intelligence. It independently evolved the largest and most complex nervous system and sophisticated behaviors among invertebrates in a separate evolutionary lineage. It is equipped with unusual traits that confer it an incredible evolutionary success: arms capable of a wide range of movements with no skeletal support; developed eyes with a complex visual behavior; vestibular system; primitive "hearing" system; chemoreceptors located in epidermis, suckers, and mouth; and a discrete olfactory organ. As if these were not enough, the occurrence of recently discovered adult neurogenesis and the high level of RNA editing give it a master key to face environmental challenges. Here we provide an overview of some of the winning evolutionary inventions that octopus puts in place such as the capacity to see color, smell by touch, edit own genes, and rejuvenate own brain.

The soft capabilities of biological appendages like the arms of Octopus vulgaris and elephants' trunks have inspired roboticists to develop their robotic equivalents. Although there have been considerable efforts to replicate their morphology and behavior patterns, we are still lagging behind in replicating the dexterity and efficiency of these biological systems. This is mostly due to the lack of development and application of dynamic controllers on these robots which could exploit the morphological properties that a soft-bodied manipulator possesses. The complexity of these high-dimensional nonlinear systems has deterred the application of traditional model-based approaches. This paper provides a machine learning-based approach for the development of dynamic models for a soft robotic manipulator and a trajectory optimization method for predictive control of the manipulator in task space. To the best of our knowledge this is the first demonstration of a learned dynamic model and a derived task space controller for a soft robotic manipulator. The validation of the controller is carried out on an octopus-inspired soft manipulator simulation derived from a piecewise constant strain approximation and then experimentally on a pneumatically actuated soft manipulator. The results indicate that such an approach is promising for developing fast and accurate dynamic models for soft robotic manipulators while being applicable on a wide range of soft manipulators.

This work presents a soft manipulator for minimally invasive surgery inspired by the biological capabilities of the octopus arm. The multi-module arm is composed of three identical units, which are able to move thanks to embedded fluidic actuators that allow omnidirectional bending and elongation, typical movements of the octopus. The use of soft materials makes the arm safe, adaptable and compliant with tissues. In addition, a granular jamming-based stiffening mechanism is integrated in each module with the aim of tuning the stiffness of the manipulator and controlling the interactions with biological structures. A miniaturized camera and a pneumatic gripper have been purposely designed and integrated on the tip of the manipulator making it usable in real working conditions. This work reports the design and the fabrication process of the manipulator, the theoretical and experimental evaluation of the stiffness and the analysis of the motion workspace. Finally, pick and place tests with the fully integrated system are shown.

Adhesion strategies that rely on mechanical interlocking or molecular attractions between surfaces can suffer when coming into contact with liquids. Thus far, artificial wet and dry adhesives have included hierarchical mushroom-shaped or porous structures that allow suction or capillarity, supramolecular structures comprising nanoparticles, and chemistry-based attractants that use various protein polyelectrolytes. However, it is challenging to develop adhesives that are simple to make and also perform well-and repeatedly-under both wet and dry conditions, while avoiding non-chemical contamination on the adhered surfaces. Here we present an artificial, biologically inspired, reversible wet/dry adhesion system that is based on the dome-like protuberances found in the suction cups of octopi. To mimic the architecture of these protuberances, we use a simple, solution-based, air-trap technique that involves fabricating a patterned structure as a polymeric master, and using it to produce a reversed architecture, without any sophisticated chemical syntheses or surface modifications. The micrometre-scale domes in our artificial adhesive enhance the suction stress. This octopus-inspired system exhibits strong, reversible, highly repeatable adhesion to silicon wafers, glass, and rough skin surfaces under various conditions (dry, moist, under water and under oil). To demonstrate a potential application, we also used our adhesive to transport a large silicon wafer in air and under water without any resulting surface contamination.

We introduce an octopus-inspired, underwater, soft-bodied robot capable of performing waterborne pulsed-jet propulsion and benthic legged-locomotion. This vehicle consists for as much as 80% of its volume of rubber-like materials so that structural flexibility is exploited as a key element during both modes of locomotion. The high bodily softness, the unconventional morphology and the non-stationary nature of its propulsion mechanisms require dynamic characterization of this robot to be dealt with by ad hoc techniques. We perform parameter identification by resorting to a hybrid optimization approach where the characterization of the dual ambulatory strategies of the robot is performed in a segregated fashion. A least squares-based method coupled with a genetic algorithm-based method is employed for the swimming and the crawling phases, respectively. The outcomes bring evidence that compartmentalized parameter identification represents a viable protocol for multi-modal vehicles characterization. However, the use of static thrust recordings as the input signal in the dynamic determination of shape-changing self-propelled vehicles is responsible for the critical underestimation of the quadratic drag coefficient.

Octopuses of the genus Cistopus Gray, 1849 are commercially valuable catches in the cephalopod fisheries of India. The primary and unique diagnostic character of this genus is the possession of eight small mucous pouches embedded in the oral faces of the webs between the bases of each arm. Historically only a single species of Cistopus, C. indicus, had been reported from Indian waters. In reviewing the octopod fauna off the Kerala coast, we have detected three species of Cistopus, of which one is described here as a new species. Cistopus platinoidus sp. nov. is distinct from Cistopus species described to date (C. indicus, C. taiwanicus and C. chinensis) on the basis of sucker counts, the number and position of enlarged suckers in males, and presence/absence of a calamus. Our studies of catch composition of Kerala octopod fisheries indicate a higher diversity of target species than previously suspected, including a number of undescribed species. Taxonomic resolution and collation of biological and distributional data are required for effective monitoring and management of these valuable fisheries.

Coleoid cephalopods (octopus, squid and cuttlefish) are active, resourceful predators with a rich behavioural repertoire. They have the largest nervous systems among the invertebrates and present other striking morphological innovations including camera-like eyes, prehensile arms, a highly derived early embryogenesis and a remarkably sophisticated adaptive colouration system. To investigate the molecular bases of cephalopod brain and body innovations, we sequenced the genome and multiple transcriptomes of the California two-spot octopus, Octopus bimaculoides. We found no evidence for hypothesized whole-genome duplications in the octopus lineage. The core developmental and neuronal gene repertoire of the octopus is broadly similar to that found across invertebrate bilaterians, except for massive expansions in two gene families previously thought to be uniquely enlarged in vertebrates: the protocadherins, which regulate neuronal development, and the C2H2 superfamily of zinc-finger transcription factors. Extensive messenger RNA editing generates transcript and protein diversity in genes involved in neural excitability, as previously described, as well as in genes participating in a broad range of other cellular functions. We identified hundreds of cephalopod-specific genes, many of which showed elevated expression levels in such specialized structures as the skin, the suckers and the nervous system. Finally, we found evidence for large-scale genomic rearrangements that are closely associated with transposable element expansions. Our analysis suggests that substantial expansion of a handful of gene families, along with extensive remodelling of genome linkage and repetitive content, played a critical role in the evolution of cephalopod morphological innovations, including their large and complex nervous systems.

Over thirty years ago anecdotal accounts of the undescribed Larger Pacific Striped Octopus suggested behaviors previously unknown for octopuses. Beak-to-beak mating, dens shared by mating pairs, inking during mating and extended spawning were mentioned in publications, and enticed generations of cephalopod biologists. In 2012-2014 we were able to obtain several live specimens of this species, which remains without a formal description. All of the unique behaviors listed above were observed for animals in aquaria and are discussed here. We describe the behavior, body color patterns, and postures of 24 adults maintained in captivity. Chromatophore patterns of hatchlings are also shown.

The cephalopod vertical lobe is the largest learning and memory structure known in invertebrate nervous systems. It is part of the visual learning circuit of the central brain, which also includes the superior frontal and subvertical lobes. Despite the well-established functional importance of this system, little is known about neuropil organization of these structures and there is to date no evidence that the five longitudinal gyri of the vertical lobe, perhaps the most distinctive morphological feature of the octopus brain, differ in their connections or molecular identities. We studied the histochemical organization of these structures in hatchling and adult Octopus bimaculoides brains with immunostaining for serotonin, octopus gonadotropin-releasing hormone (oGNRH), and octopressin-neurophysin (OP-NP). Our major finding is that the five lobules forming the vertical lobe gyri have distinct neurochemical signatures. This is most prominent in the hatchling brain, where the median and mediolateral lobules are enriched in OP-NP fibers, the lateral lobule is marked by oGNRH innervation, and serotonin immunostaining heavily labels the median and lateral lobules. A major source of input to the vertical lobe is the superior frontal lobe, which is dominated by a neuropil of interweaving fiber bundles. We have found that this neuropil also has an intrinsic neurochemical organization: it is partitioned into territories alternately enriched or impoverished in oGNRH-containing fascicles. Our findings establish that the constituent lobes of the octopus superior frontal-vertical system have an intricate internal anatomy, one likely to reflect the presence of functional subsystems within cephalopod learning circuitry., (© 2015 Wiley Periodicals, Inc.)

Octopus suckers are able to attach to all nonporous surfaces and generate a very strong attachment force. The well-known attachment features of this animal result from the softness of the sucker tissues and the surface morphology of the portion of the sucker that is in contact with objects or substrates. Unlike artificial suction cups, octopus suckers are characterized by a series of radial grooves that increase the area subjected to pressure reduction during attachment. In this study, we constructed artificial suction cups with different surface geometries and tested their attachment performances using a pull-off setup. First, smooth suction cups were obtained for casting; then, sucker surfaces were engraved with a laser cutter. As expected, for all the tested cases, the engraving treatment enhanced the attachment performance of the elastomeric suction cups compared with that of the smooth versions. Moreover, the results indicated that the surface geometry with the best attachment performance was the geometry most similar to octopus sucker morphology. The results obtained in this work can be utilized to design artificial suction cups with higher wet attachment performance.

Suction cups are often found in nature as attachment strategy in water. Nevertheless, the application of the artificial counterpart is limited by the dimension of the actuators and their usability in wet conditions. A novel design for the development of a suction cup inspired by octopus suckers is presented. The main focus of this research was on the modelling and characterization of the actuation unit, and a first prototype of the suction cup was realized as a proof of concept. The actuation of the suction cup is based on dielectric elastomer actuators. The presented device works in a wet environment, has an integrated actuation system, and is soft. The dimensions of the artificial suction cups are comparable to proximal octopus suckers, and the attachment mechanism is similar to the biological counterpart. The design approach proposed for the actuator allows the definition of the parameters for its development and for obtaining a desired pressure in water. The fabricated actuator is able to produce up to 6 kPa of pressure in water, reaching the maximum pressure in less than 300 ms.

Understanding the response of any species to climate change can be challenging. However, in short-lived species the faster turnover of generations may facilitate the examination of responses associated with longer-term environmental change. Octopus tetricus, a commercially important species, has undergone a recent polewards range shift in the coastal waters of south-eastern Australia, thought to be associated with the southerly extension of the warm East Australian Current. At the cooler temperatures of a polewards distribution limit, growth of a species could be slower, potentially leading to a bigger body size and resulting in a slower population turnover, affecting population viability at the extreme of the distribution. Growth rates, body size, and life span of O. tetricus were examined at the leading edge of a polewards range shift in Tasmanian waters (40°S and 147°E) throughout 2011. Octopus tetricus had a relatively small body size and short lifespan of approximately 11 months that, despite cooler temperatures, would allow a high rate of population turnover and may facilitate the population increase necessary for successful establishment in the new extended area of the range. Temperature, food availability and gender appear to influence growth rate. Individuals that hatched during cooler and more productive conditions, but grew during warming conditions, exhibited faster growth rates and reached smaller body sizes than individuals that hatched into warmer waters but grew during cooling conditions. This study suggests that fast growth, small body size and associated rapid population turnover may facilitate the range shift of O. tetricus into Tasmanian waters.

Despite extensive revisions over recent decades, the taxonomy of benthic octopuses (Family Octopodidae) remains in a considerable flux. Among groups of unresolved status is a species complex of morphologically similar shallow-water octopods from subtropical Australasia, including: Allopatric populations of Octopus tetricus on the eastern and western coasts of Australia, of which the Western Australian form is speculated to be a distinct or sub-species; and Octopus gibbsi from New Zealand, a proposed synonym of Australian forms. This study employed a combination of molecular and morphological techniques to resolve the taxonomic status of the 'tetricus complex'. Phylogenetic analyses (based on five mitochondrial genes: 12S rRNA, 16S rRNA, COI, COIII and Cytb) and Generalised Mixed Yule Coalescent (GMYC) analysis (based on COI, COIII and Cytb) distinguished eastern and Western Australian O. tetricus as distinct species, while O. gibbsi was found to be synonymous with the east Australian form (BS = >97, PP = 1; GMYC p = 0.01). Discrete morphological differences in mature male octopuses (based on sixteen morphological traits) provided further evidence of cryptic speciation between east (including New Zealand) and west coast populations; although females proved less useful in morphological distinction among members of the tetricus complex. In addition, phylogenetic analyses suggested populations of octopuses currently treated under the name Octopus vulgaris are paraphyletic; providing evidence of cryptic speciation among global populations of O. vulgaris, the most commercially valuable octopus species worldwide.

A major component of cephalopod adaptive camouflage behavior has rarely been studied: their ability to change the three-dimensionality of their skin by morphing their malleable dermal papillae. Recent work has established that simple, conical papillae in cuttlefish (Sepia officinalis) function as muscular hydrostats; that is, the muscles that extend a papilla also provide its structural support. We used brightfield and scanning electron microscopy to investigate and compare the functional morphology of nine types of papillae of different shapes, sizes and complexity in six species: S. officinalis small dorsal papillae, Octopus vulgaris small dorsal and ventral eye papillae, Macrotritopus defilippi dorsal eye papillae, Abdopus aculeatus major mantle papillae, O. bimaculoides arm, minor mantle, and dorsal eye papillae, and S. apama face ridge papillae. Most papillae have two sets of muscles responsible for extension: circular dermal erector muscles arranged in a concentric pattern to lift the papilla away from the body surface and horizontal dermal erector muscles to pull the papilla's perimeter toward its core and determine shape. A third set of muscles, retractors, appears to be responsible for pulling a papilla's apex down toward the body surface while stretching out its base. Connective tissue infiltrated with mucopolysaccharides assists with structural support. S. apama face ridge papillae are different: the contraction of erector muscles perpendicular to the ridge causes overlying tissues to buckle. In this case, mucopolysaccharide-rich connective tissue provides structural support. These six species possess changeable papillae that are diverse in size and shape, yet with one exception they share somewhat similar functional morphologies. Future research on papilla morphology, biomechanics and neural control in the many unexamined species of octopus and cuttlefish may uncover new principles of actuation in soft, flexible tissue.

Cholinergic structures in the arm of the cephalopod Octopus vulgaris were studied by immunohistochemistry using specific antisera for two types (common and peripheral) of acetylcholine synthetic enzyme choline acetyltransferase (ChAT): antiserum raised against the rat common type ChAT (cChAT), which is cross-reactive with molluscan cChAT, and antiserum raised against the rat peripheral type ChAT (pChAT), which has been used to delineate peripheral cholinergic structures in vertebrates, but not previously in invertebrates. Western blot analysis of octopus extracts revealed a single pChAT-positive band, suggesting that pChAT antiserum is cross-reactive with an octopus counterpart of rat pChAT. In immunohistochemistry, only neuronal structures of the octopus arm were stained by cChAT and pChAT antisera, although the pattern of distribution clearly differed between the two antisera. cChAT-positive varicose nerve fibers were observed in both the cerebrobrachial tract and neuropil of the axial nerve cord, while pChAT-positive varicose fibers were detected only in the neuropil of the axial nerve cord. After epitope retrieval, pChAT-positive neuronal cells and their processes became visible in all ganglia of the arm, including the axial and intramuscular nerve cords, and in ganglia of suckers. Moreover, pChAT-positive structures also became detectable in nerve fibers connecting the different ganglia, in smooth nerve fibers among muscle layers and dermal connective tissues, and in sensory cells of the suckers. These results suggest that the octopus arm has two types of cholinergic nerves: cChAT-positive nerves from brain ganglia and pChAT-positive nerves that are intrinsic to the arm.

In the present study, we determined the complete mitochondrial DNA (mtDNA) sequences of two species of Cistopus, namely C. chinensis and C. taiwanicus, and conducted a comparative mt genome analysis across the class Cephalopoda. The mtDNA length of C. chinensis and C. taiwanicus are 15706 and 15793 nucleotides with an AT content of 76.21% and 76.5%, respectively. The sequence identity of mtDNA between C. chinensis and C. taiwanicus was 88%, suggesting a close relationship. Compared with C. taiwanicus and other octopods, C. chinensis encoded two additional tRNA genes, showing a novel gene arrangement. In addition, an unusual 23 poly (A) signal structure is found in the ATP8 coding region of C. chinensis. The entire genome and each protein coding gene of the two Cistopus species displayed notable levels of AT and GC skews. Based on sliding window analysis among Octopodiformes, ND1 and DN5 were considered to be more reliable molecular beacons. Phylogenetic analyses based on the 13 protein-coding genes revealed that C. chinensis and C. taiwanicus form a monophyletic group with high statistical support, consistent with previous studies based on morphological characteristics. Our results also indicated that the phylogenetic position of the genus Cistopus is closer to Octopus than to Amphioctopus and Callistoctopus. The complete mtDNA sequence of C. chinensis and C. taiwanicus represent the first whole mt genomes in the genus Cistopus. These novel mtDNA data will be important in refining the phylogenetic relationships within Octopodiformes and enriching the resource of markers for systematic, population genetic and evolutionary biological studies of Cephalopoda.

The prevalence of coccidian parasites in three Octopus tehuelchus populations from San Matías Gulf (Patagonia, Argentina) is compared. The prevalence was similar between sexes, but varied between seasons (being highest during cold months) and sites. Islote Lobos had the highest prevalence (42.7-100%) followed by San Antonio Bay (0-66%) and El Fuerte (0-24.5%). Octopuses under 27 mm of dorsal mantle length showed a low prevalence (less than 50%), which increased with size. We hypothesize that the high prevalence of parasites, which affect the three populations differentially, could account for the observed variability in life-span and growth, size-frequency distributions, reproduction and densities of O. tehuelchus populations., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The octopus sucker represents a fascinating natural system performing adhesion on different terrains and substrates. Octopuses use suckers to anchor the body to the substrate or to grasp, investigate and manipulate objects, just to mention a few of their functions. Our study focuses on the morphology and adhesion mechanism of suckers in Octopus vulgaris. We use three different techniques (MRI, ultrasonography, and histology) and a 3D reconstruction approach to contribute knowledge on both morphology and functionality of the sucker structure in O. vulgaris. The results of our investigation are two-fold. First, we observe some morphological differences with respect to the octopus species previously studied (i.e., Octopus joubini, Octopus maya, Octopus bimaculoides/bimaculatus and Eledone cirrosa). In particular, in O. vulgaris the acetabular chamber, that is a hollow spherical cavity in other octopuses, shows an ellipsoidal cavity which roof has an important protuberance with surface roughness. Second, based on our findings, we propose a hypothesis on the sucker adhesion mechanism in O. vulgaris. We hypothesize that the process of continuous adhesion is achieved by sealing the orifice between acetabulum and infundibulum portions via the acetabular protuberance. We suggest this to take place while the infundibular part achieves a completely flat shape; and, by sustaining adhesion through preservation of sucker configuration. In vivo ultrasonographic recordings support our proposed adhesion model by showing the sucker in action. Such an underlying physical mechanism offers innovative potential cues for developing bioinspired artificial adhesion systems. Furthermore, we think that it could possibly represent a useful approach in order to investigate any potential difference in the ecology and in the performance of adhesion by different species.

Cephalopods such as the octopus show the most advanced behavior among invertebrates, which they accomplish with an exceptionally flexible body plan. In this review I propose that the embodied organization approach, developed by roboticists to design efficient autonomous robots, is useful for understanding the evolution and development of the efficient adaptive interaction of animals with their environment, using the octopus as the leading example. The embodied organization approach explains adaptive behavior as emerging from the continuous dynamical and reciprocal physical and informational interactions between four elements: the controller, the mechanical and the sensory systems and the environment. In contrast to hierarchical organization, in embodied organization, self-organization processes can take part in the emergence of the adaptive properties. I first discuss how the embodiment concept explains covariation of body form, nervous system organization, and level of behavioral complexity using the Mollusca as an example. This is an ideal phylum to test such a qualitative correlation between body/brain/behavior, because they show the greatest variations of body plan within a single phylum. In some cases the covariation of nervous system and body structure seems to arise independently of close phylogenetic relationships. Next, I dwell on the octopus as an ideal model to test the embodiment concept within a single biological system. Here, the unusual body morphology of the octopus exposes the uniqueness of the four components comprising the octopus' embodiment. Considering together the results from behavioral, physiological, anatomical, and motor control research suggests that these four elements mutually influence each other. It is this mutual interactions and self-organization which have led to their unique evolution and development to create the unique and highly efficient octopus embodiment., (© 2013 S. Karger AG, Basel.)

Vampire squid (Vampyroteuthis infernalis) are considered phylogenetic relics with cephalopod features of both octopods and squids. They lack feeding tentacles, but in addition to their eight arms, they have two retractile filaments, the exact functions of which have puzzled scientists for years. We present the results of investigations on the feeding ecology and behaviour of Vampyroteuthis, which include extensive in situ, deep-sea video recordings from MBARI's remotely operated vehicles (ROVs), laboratory feeding experiments, diet studies and morphological examinations of the retractile filaments, the arm suckers and cirri. Vampire squid were found to feed on detrital matter of various sizes, from small particles to larger marine aggregates. Ingested items included the remains of gelatinous zooplankton, discarded larvacean houses, crustacean remains, diatoms and faecal pellets. Both ROV observations and laboratory experiments led to the conclusion that vampire squid use their retractile filaments for the capture of food, supporting the hypothesis that the filaments are homologous to cephalopod arms. Vampyroteuthis' feeding behaviour is unlike any other cephalopod, and reveals a unique adaptation that allows these animals to spend most of their life at depths where oxygen concentrations are very low, but where predators are few and typical cephalopod food is scarce.

Octopuses have a unique flexible body and unusual morphology, but nevertheless they are undoubtedly a great evolutionary success. They compete successfully with vertebrates in their ecological niche using a rich behavioral repertoire more typical of an intelligent predator which includes extremely effective defensive behavior--fast escape swimming and an astonishing ability to adapt their shape and color to their environment. The most obvious characteristic feature of an octopus is its eight long and flexible arms, but these pose a great challenge for achieving the level of motor and sensory information processing necessary for their behaviors. First, coordinating motion is a formidable task because of the infinite degrees of freedom that have to be controlled; and second, it is hard to use body coordinates in this flexible animal to represent sensory information in a central control system. Here I will review experimental results suggesting that these difficulties, arising from the animal's morphology, have imposed the evolution of unique brain/body/behavior relationships best explained as intelligent behavior which emerges from the octopus's embodied organization. The term 'intelligent embodiment' comes from robotics and refers to an approach to designing autonomous robots in which the behavior emerges from the dynamic physical and sensory interactions of the agent's materials, morphology and environment. Consideration of the unusual neurobiology of the octopus in the light of its unique morphology suggests that similar embodied principles are instrumental for understanding the emergence of intelligent behavior in all biological systems., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The present study contributes to the question of school literacy about the brain, with an original survey conducted on Italian students from the 3(rd) to 10(th) grades (n=508). The main goal was to test student's knowledge, attitudes, and interests about neuroscience, to assess needs, prospects, and difficulties in teaching about the brain from elementary to high school. A written questionnaire, maintaining anonymity, asked 12 close-ended multiple choice questions on topics related to human and animal brains, plus one facultative open-ended question about interests and curiosities on brain topics. The results show that respondents have a fragmentary level of basic knowledge about the brain, with aspects related to brain functions and consciousness the most challenging. As expected, degrees of performance improve with school level; elementary school students answered correctly an average number of 5.3 questions, middle school 6.5, and high school 7.4. Overall, students show great interest in the brain, as shown by the large number of questions gathered through the open-ended question (n=384). Other topics are addressed, mostly related to brain structure/functions and the role of the brain in the everyday life. The survey indicates the need of more thorough school programs on this subject, reinforced by interdisciplinary teaching where comparative anatomy and evolutionary aspects of brain development are covered.

Octopus arms are extremely dexterous structures. The special arrangements of the muscle fibers and nerve cord allow a rich variety of complex and fine movements under neural control. Historically, the arm structure has been investigated using traditional comparative morphological ex vivo analysis. Here, we employed ultrasound imaging, for the first time, to explore in vivo the arms of the cephalopod mollusc Octopus vulgaris. Sonographic examination (linear transducer, 18 MHz) was carried out in anesthetized animals along the three anatomical planes: transverse, sagittal and horizontal. Images of the arm were comparable to the corresponding histological sections. We were able, in a non-invasive way, to measure the dimensions of the arm and its internal structures such as muscle bundles and neural components. In addition, we evaluated echo intensity signals as an expression of the difference in the muscular organization of the tissues examined (i.e. transverse versus longitudinal muscles), finding different reflectivity based on different arrangements of fibers and their intimate relationship with other tissues. In contrast to classical preparative procedures, ultrasound imaging can provide rapid, destruction-free access to morphological data from numerous specimens, thus extending the range of techniques available for comparative studies of invertebrate morphology.

Background: To what extent are the properties of neuronal networks constrained by computational considerations? Comparative analysis of the vertical lobe (VL) system, a brain structure involved in learning and memory, in two phylogenetically close cephalopod mollusks, Octopus vulgaris and the cuttlefish Sepia officinalis, provides a surprising answer to this question., Results: We show that in both the octopus and the cuttlefish the VL is characterized by the same simple fan-out fan-in connectivity architecture, composed of the same three neuron types. Yet, the sites of short- and long-term synaptic plasticity and neuromodulation are different. In the octopus, synaptic plasticity occurs at the fan-out glutamatergic synaptic layer, whereas in the cuttlefish plasticity is found at the fan-in cholinergic synaptic layer., Conclusions: Does this dramatic difference in physiology imply a difference in function? Not necessarily. We show that the physiological properties of the VL neurons, particularly the linear input-output relations of the intermediate layer neurons, allow the two different networks to perform the same computation. The convergence of different networks to the same computational capacity indicates that it is the computation, not the specific properties of the network, that is self-organized or selected for by evolutionary pressure., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Many aspects of octopus growth dynamics are poorly understood, particularly in relation to sub-adult or adult growth, muscle fibre dynamics and repro-somatic investment. The growth of 5 month old Octopus pallidus cultured in the laboratory was investigated under three temperature regimes over a 12 week period: seasonally increasing temperatures (14-18°C); seasonally decreasing temperatures (18-14°C); and a constant temperature mid-way between seasonal peaks (16°C). Differences in somatic growth at the whole-animal level, muscle tissue structure and rate of gonad development were investigated. Continuous exponential growth was observed, both at a group and at an individual level, and there was no detectable effect of temperature on whole-animal growth rate. Juvenile growth rate (from 1 to 156 days) was also monitored prior to the controlled experiment; exponential growth was observed, but at a significantly faster rate than in the older experimental animals, suggesting that O. pallidus exhibit a double-exponential two-phase growth pattern. There was considerable variability in size-at-age even between individuals growing under identical thermal regimes. Animals exposed to seasonally decreasing temperatures exhibited a higher rate of gonad development compared with animals exposed to increasing temperatures; however, this did not coincide with a detectable decline in somatic growth rate or mantle condition. The ongoing production of new mitochondria-poor and mitochondria-rich muscle fibres (hyperplasia) was observed, indicated by a decreased or stable mean muscle fibre diameter concurrent with an increase in whole-body size. Animals from both seasonal temperature regimes demonstrated higher rates of new mitochondria-rich fibre generation relative to those from the constant temperature regime, but this difference was not reflected in a difference in growth rate at the whole-body level. This is the first study to record ongoing hyperplasia in the muscle tissue of an octopus species, and provides further insight into the complex growth dynamics of octopus.

Information regarding melatonin production in molluscs is very limited. In this study the presence and daily fluctuations of melatonin levels were investigated in hemolymph, retina and nervous system-related structures in the cephalopod Octopus vulgaris. Adult animals were maintained in captivity under natural photoperiod and killed at different times in a regular daily cycle. Levels of melatonin, serotonin (5-HT) and its acid metabolite (5-hydroxyindole acetic acid, 5-HIAA) in the hemolymph, retina, optic lobe, and cerebral ganglion were assayed by HPLC. Melatonin content fluctuated rhythmically in the retina and hemolymph, peaking at night. In the retina, but not in the other neural tissues, the rhythm was opposite to that of 5-HT, which displayed basal levels at night. Also, 5-HIAA levels in the retina were higher during the night, supporting that rhythmic melatonin production could be linked to diurnal changes in 5-HT degradation. The high levels of melatonin found in the retina point to it as the major source of melatonin in octopus; in addition, a large variation of melatonin content was found in the optic lobe with maximal values at night. All these data suggest that melatonin might play a role in the transduction of the light-dark cycle information for adjustment of rhythmic physiological events in cephalopods.