A model to explain spermatophore implantation in cephalopods (Mollusca: Cephalopoda) and a discussion on its evolutionary origins and significance

Male coleoid cephalopods produce spermatophores that can attach autonomously on the female's body during a complex process of evagination called the spermatophoric reaction, during which the ejaculatory apparatus and spiral filament of the spermatophore are everted and exposed to the external milieu. In some deepwater cephalopods, the reaction leads to the intradermal implantation of the spermatophore, a hitherto enigmatic phenomenon. The present study builds upon several lines of evidence to propose that spermatophore implantation is probably achieved through the combination of (1) an evaginating-tube mechanism performed by the everting ejaculatory apparatus and (2) the anchorage provided by the spiral filament's stellate particles. The proposed theoretical model assumes that, as it is exposed to the external milieu, each whorl of the spiral filament anchors to the surrounding tissue by means of its sharp stellate particles. As the ejaculatory apparatus tip continues evaginating, it grows in diameter and stretches lengthwise, enlarging the diameter of the whorl and propelling it, consequently tearing and pushing the anchored tissue outward and backward, and opening space for the next whorl to attach. After the ejaculatory apparatus has been everted and has perforated tissue, the cement body is extruded, possibly aiding in final attachment, and the sperm mass comes to lie inside the female tissue, encompassed by the everted ejaculatory apparatus tube. It is proposed that this unique, efficient spermatophore attachment mechanism possibly evolved in intimate relationship with the adoption of an active mode of life by coleoids. The possible roles of predation pressure and sperm competition in the evolution of this mechanism are also discussed. (c) 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105, 711726.

O enigma da "reação espermatofórica": breve síntese do conhecimento sobre a estrutura e o funcionamento dos espermatóforos dos cefalópodes (Mollusca: Cephalopoda)

Cefalópodes coleóides (lulas, sépias e polvos) produzem espermatóforos muito complexos que são transferidos à fêmea durante a cópula por meio do hectocótilo, um apêndice modificado nos machos. Durante a transferência à fêmea, ocorre a chamada "reação espermatofórica", complexo processo de evaginação do aparato ejaculatório do espermatóforo, que conduz à exteriorização da massa espermática e corpo cimentante. A presente revisão sintetiza o conhecimento acerca da morfologia e funcionamento desta estrutura exclusiva dos coleóides, identificando lacunas e definindo estratégias que possibilitem avanços na área. Poucos trabalhos abordam com detalhes a morfologia e anatomia funcional dos espermatóforos dos cefalópodes, grande parte do conhecimento acerca da estrutura do espermatóforo tendo sido gerada por trabalhos clássicos do século XIX e início do século XX. Investigações acerca do funcionamento dos espermatóforos são consideravelmente mais raras, estando o conhecimento básico sobre a reação espermatofórica restrito a apenas 19 espécies de coleóides. A revisão da literatura especializada permite sugerir que existem dois tipos básicos de fixação de espermatóforos em Decapodiformes (lulas e sepióides): fixação superficial e implante profundo (ou intra-dérmico). Na fixação superficial, comum em diversas espécies (e.g., Loliginidae, Sepiidae, Ommastrephidae), a base dos espermatângios é aderida ao tecido-alvo aparentemente por meio do corpo cimentante, a partir de substâncias adesivas e, em alguns casos, estruturas de fixação. No implante profundo, comum em alguns grupos de lulas oceânicas e de águas profundas (e.g., Architeuthidae, Cranchiidae, Octopoteuthidae, Sepiolidae), os espermatóforos implantam-se inteiramente no corpo da fêmea, de forma autônoma. Permanece desconhecido o mecanismo responsável pelo implante profundo. Em Octopodiformes (polvos), o espermatóforo é inserido no gonoduto feminino, alcançando a glândula oviducal, onde estão localizadas as espermatecas, ou a cavidade do ovário. Como o funcionamento extracorpóreo dos espermatóforos depende exclusivamente da intrincada estrutura e organização de seus componentes (e.g., membranas e túnicas), somente investigações detalhadas dessas estruturas proverão as bases para a compreensão do funcionamento e da exata função do complexo espermatóforo dos coleóides. Recomenda-se o desenvolvimento de um protocolo simples e eficiente para coloração e preparação total de espermatóforos, de forma que seja possível expandir as descrições morfológicas do espermatóforo em estudos taxonômicos e anatômicos, permitindo, portanto, ampliação do conhecimento acerca desta enigmática estrutura.

Spermatophoric reaction reappraised: Novel insights into the functioning of the loliginid spermatophore based on Doryteuthis plei (Mollusca: Cephalopoda)

During copulation, spermatophores produced by male coleoid cephalopods undergo the spermatophoric reaction, a complex process of evagination that culminates in the attachment of the spermatangium (everted spermatophore containing the sperm mass) on the female's body. To better understand this complicated phenomenon, the present study investigated the functional morphology of the spermatophore of the squid Doryteuthis plei applying in vitro analysis of the reaction, as well as light and electron microscopy investigation of spermatangia obtained either in vitro, or naturally attached on females. Hitherto unnoticed functional features of the loliginid spermatophore require a reappraisal of some important processes involved in the spermatophoric reaction. The most striking findings concern the attachment mechanism, which is not carried out solely by cement adhesive material, as previously believed, but rather by an autonomous, complex process performed by multiple structures during the spermatophoric reaction. During evagination, the ejaculatory apparatus provides anchorage on the targeted tissue, presumably due to the minute stellate particles present in the exposed spiral filament. Consequently, the ejaculatory apparatus maintains the attachment of the tip of the evaginating spermatophore until the cement body is extruded. Subsequently, the cement body passes through a complex structural rearrangement, which leads to the injection of both its viscid contents and pointed oral region onto the targeted tissue. The inner membrane at the oral region of the cement body contains numerous stellate particles attached at its inner side; eversion of this membrane exposes these sharp structures, which presumably adhere to the tissue and augment attachment. Several naturally attached spermatangia were found with their bases implanted at the deposition sites, and the possible mechanisms of perforation are discussed based on present evidence. The function of the complex squid spermatophore and its spermatophoric reaction is revisited in light of these findings. J. Morphol. 2012. (C) 2011 Wiley Periodicals, Inc.

Unraveling the structure of squids' spermatophores: a combined approach based on Doryteuthis plei (Blainville, 1823) (Cephalopoda: Loliginidae)

Marian, J.E.A.R. and Domaneschi, O. 2012. Unraveling the structure of squids spermatophores: a combined approach based on Doryteuthis plei (Blainville, 1823) (Cephalopoda: Loliginidae). Acta Zoologica (Stockholm) 93: 281307. Male coleoid cephalopods produce elaborate spermatophores, which function autonomously outside the male body during copulation, undergoing a complicated process of evagination. In order to contribute to the understanding of this unique structure, this study investigated the morphology of the spermatophore of Doryteuthis plei applying several microscopy techniques. A hitherto unreported, much more complex structural arrangement was revealed for the loliginid spermatophore, the most striking findings being: (1) the complex, layered structure of the middle membrane, which bears an additional, chemically distinct segment surrounding part of the cement body; (2) the presence of a space between the inner tunic and middle membrane filled with a fine reticulated material; (3) the presence of stellate particles not only embedded in the spiral filament, but also closely applied to the inner membrane at the level of the cement body; (4) the presence of a pre-oral chamber in the cap region; and (5) the complex organization of the cement body, formed by two distinct layers encompassing contents of different chemical and textural properties. Careful literature reassessment suggests several of these features are common to loliginids, and to some extent to other squids. Their possible functional implications are discussed in light of our knowledge of the spermatophoric reaction mechanics.

Analysis of the reproductive system of female vampire squid

Analysis of the reproductive system of female vampire squid to determine reproductive strategy and fecundity of vampire squid, accessioned in the Santa Barbara Museum of Natural History and collected in 60, 70s from off southern California.

On the microstructure, growth pattern and original porosity of belemnite rostra: insights from calcitic Jurassic belemnites

Calcitic belemnite rostra are usually employed to perform paleoenvironmental studies based on geochemical data. However, several questions, such as their original porosity and microstructure, remain open, despite they are essential to make accurate interpretations based on geochemical analyses.This paper revisits and enlightens some of these questions. Petrographic data demonstrate that calcite crystals of the rostrum solidum of belemnites grow from spherulites that successively develop along the apical line, resulting in a “regular spherulithic prismatic” microstructure. Radially arranged calcite crystals emerge and diverge from the spherulites: towards the apex, crystals grow until a new spherulite is formed; towards the external walls of the rostrum, the crystals become progressively bigger and prismatic. Adjacent crystals slightly vary in their c-axis orientation, resulting in undulose extinction. Concentric growth layering develops at different scales and is superimposed and traversed by a radial pattern, which results in the micro-fibrous texture that is observed in the calcite crystals in the rostra.Petrographic data demonstrate that single calcite crystals in the rostra have a composite nature, which strongly suggests that the belemnite rostra were originally porous. Single crystals consistently comprise two distinct zones or sectors in optical continuity: 1) the inner zone is fluorescent, has relatively low optical relief under transmitted light (TL) microscopy, a dark-grey color under backscatter electron microscopy (BSEM), a commonly triangular shape, a “patchy” appearance and relatively high Mg and Na contents; 2) the outer sector is non-fluorescent, has relatively high optical relief under TL, a light-grey color under BSEM and low Mg and Na contents. The inner and fluorescent sectors are interpreted to have formed first as a product of biologically controlled mineralization during belemnite skeletal growth and the non-fluorescent outer sectors as overgrowths of the former, filling the intra- and inter-crystalline porosity. This question has important implications for making paleoenvironmental and/or paleoclimatic interpretations based on geochemical analyses of belemnite rostra.Finally, the petrographic features of composite calcite crystals in the rostra also suggest the non-classical crystallization of belemnite rostra, as previously suggested by other authors.

SPE-LC-FD Determination of Polycyclic Aromatic Hydrocarbon Monohydroxy Derivatives in Cephalopods

A new analytical methodology, based on liquid chromatography with fluorescence detection (LC-FD), after extraction, enzymatic hydrolysis, and solid-phase extraction (SPE) through Oasis HLB cartridges, was developed and validated for the simultaneous determination of three monohydroxy derivatives of polycyclic aromatic hydrocarbons (PAHs). The optimized analytical method is sensitive, accurate, and precise, with recoveries between 62 and 110% and limits of detection of 227, 9, and 45 ng/g for 1-hydroxynaphthalene, 2-hydroxyfluorene, and 1-hydroxypyrene, respectively. Their levels were estimated in different cephalopod matrices (edible tissues and hemolymph). The methodology was applied to samples of the major cephalopod species consumed worldwide. Of the 18 samples analyzed, 39% were contaminated with 1-hydroxynaphthalene, which was the only PAH metabolite detected. Its concentration ranged from 786 to 1145 ng/g. This highly sensitive and specific method allows the identification and quantitation of PAH metabolites in forthcoming food safety and environmental monitoring programs.

Bermudan cephalopods.

v.39:no.40(1960)

Isolation and characterization of glycosaminoglycans and a study of its bioactive potential in two commercially important species of cephalopods, Loligo duvauceli and Sepia pharaonis

The present study is aimed at the isolation and characterization of glycosaminoglycans from selected tissues of two commercially important species of cephalopods;squid,Loligo duvauceli and cuttlefish,Sepia pharaonis,keeping in view of the aforementioned benefits on the utilization of waste generated during processing.The cephalopod GAGs may also be expected to have an effect on various physiological functions based on the results obtained from GAGs from other sources.In addition,knowledge of the chemical structure of macromolecules that constitute major components of extracellular matrix(ECM) will be helpful in understanding their interactions with other matrix components.

Studies on Some Aspects of Landings,Utilization and Export of Commercially Important Cephalopods

In the present study two aspects of cephalopod fishery such as (i) landings and (ii) utilization including export of cephalopods from Indian are dealt with. Under the topic landings, craft, both traditional and mechanized and fishing techniques employed for capturing cephalopods are discussed. General trend in cephalopod landing in India and particularly south west coast, species composition, description and geographic distribution of important species of cuttle fishes, squids and octopus and the distribution and abundance of cephalopods along the continental shelf of the south west coast are also discussed. Some biological aspects like population parameters, maturation and spawning, food and feeding of the two representive species of cuttle fishes and squid viz, Sepia pharaonis and Loligo duvauceli are studied.Under utilization, general handling and processing of various frozen cephalopods products for exports and commercial operation in a typical plant are studied. The export of cephalopod products since its commencement is studied in detail and market strategy for enhancing the export are suggested.

Are deep-sea cephalopods really common preys for oceanic seabirds?

Uma análise de dados publicados sobre dietas de aves marinhas oceânicas mostra a predominância de cefalópodes musculares e de distribuição mais superficial nas camadas oceânicas, mas também são importantes as espécies gelatinosas e amoniacais restritas a camadas abaixo dos 300 m da superfície. A princípio, não deveria se esperar que cefalópodes de profundidade fossem considerados presas comuns de aves marinhas oceânicas como reportados por muitos autores. É proposto neste estudo que uma fonte indireta, importante e de fácil obtenção, surgiu com o início das atividades dos barcos atuneiros que operam com espinhel. O hábito de ingerir restos de vísceras de peixes capturados em barcos espinheleiros pode explicar as prováveis conclusões equivocadas de que cefalópodes de profundidade são presas naturais de aves marinhas oceânicas.

Localization of ion-regulatory epithelia in embryos and hatchlings of two cephalopods

The tissue distribution and ontogeny of Na+/K+-ATPase has been examined as an indicator for ion-regulatory epithelia in whole animal sections of embryos and hatchlings of two cephalopod species: the squid Loligo vulgaris and the cuttlefish Sepia officinalis. This is the first report of the immunohistochemical localization of cephalopod Na+/K+-ATPase with the polyclonal antibody alpha (H-300) raised against the human alpha1-subunit of Na+/K+-ATPase. Na+/K+-ATPase immunoreactivity was observed in several tissues (gills, pancreatic appendages, nerves), exclusively located in baso-lateral membranes lining blood sinuses. Furthermore, large single cells in the gill of adult L. vulgaris specimens closely resembled Na+/K+-ATPase-rich cells described in fish. Immunohistochemical observations indicated that the amount and distribution of Na+/K+-ATPase in late cuttlefish embryos was similar to that found in juvenile and adult stages. The ion-regulatory epithelia (e.g., gills, excretory organs) of the squid embryos and paralarvae exhibited less differentiation than adults. Na+/K+-ATPase activities for whole animals were higher in hatchlings of S. officinalis (157.0 ± 32.4 µmol/g FM/h) than in those of L. vulgaris (31.8 ± 3.3 µmol/g FM/h). S. officinalis gills and pancreatic appendages achieved activities of 94.8 ± 18.5 and 421.8 ± 102.3 µmol ATP/g FM/h, respectively. High concentrations of Na+/K+-ATPase in late cephalopod embryos might be important in coping with the challenging abiotic conditions (low pH, high pCO2) that these organisms encounter inside their eggs. Our results also suggest a higher sensitivity of squid vs. cuttlefish embryos to environmental acid-base disturbances.