Light and electron microscopy study of the salivary glands of the carnivorous opisthobranch Philinopsis depicta (Mollusca, Gastropoda)

Cephalaspideans are a group of opisthobranch gastropods that comprises carnivorous and herbivorous species, allowing an investigation of the relationship between these diets and the morphofunctional features of the salivary glands. In this study, the salivary glands of the carnivorous cephalaspidean Philinopsis depicta were observed by light and electron microscopy. The secretory epithelium of these ribbon-shaped glands is formed by ciliated cells, granular cells and cells with apical vacuole. In ciliated cells the nucleus and most cytoplasmic organelles are located in the wider apical region and a very thin stalk reaches the base of the epithelium. These cells possess significant amounts of glycogen. Granular cells are packed with electron-dense secretory granules and also contain several cisternae of rough endoplasmic reticulum and Golgi stacks. The other type of secretory cell is mainly characterized by the presence of a large apical vacuole containing secretion. These cells possess high amounts of rough endoplasmic reticulum cisternae and several Golgi stacks. Vesicles with peripheral electron-dense material are also abundant, and seem to fuse to form the apical vacuole. The available data point out to a significant difference between the salivary glands of carnivorous and herbivorous cephalaspidean opisthobranchs, with an intensification of protein secretion in carnivorous species.     

Light and electron microscopic study of the anterior oesophagus of Bulla striata (Mollusca,Opisthobranchia)

Lobo‐da‐Cunha, A., Oliveira, E., Alves, Â., Coelho, R. and Calado, G. 2010. Light and electron microscopic study of the anterior oesophagus of Bulla striata (Mollusca, Opisthobranchia). —Acta Zoologica (Stockholm) 91 : 125–138. The anterior oesophagus of Bulla striata was investigated with light and electron microscopy. In the most anterior region, the ridges of the oesophageal wall are covered by a ciliated columnar epithelium forming large apical blebs which are released into the lumen, an activity that is particularly intense in the oesophageal pouch. In the last two‐thirds of the anterior oesophagus, the epithelium is covered with microvilli embedded in a cuticle, but apocrine secretion and cilia are absent. Subepithelial secretory cells are very abundant in the oesophageal wall, except in the roof of the pouch. They have a long neck that crosses the epithelium, whereas the cell body containing the nucleus is embedded in the connective tissue. Large electron‐lucent secretory vesicles and many Golgi stacks fill most of their cytoplasm. The histochemical and cytochemical assays show that these cells secrete acid mucopolysaccharides. With the current and future studies we aim to obtain data for the establishment of relationships between morphofunctional features of the digestive system and food types in cephalaspideans. Additionally, the new data about the oesophageal pouch of B. striata may be useful for the establishment of eventual homologies with the oesophageal diverticula of other opisthobranchs.     

Larval development of Balanus reticulatus Utinomi, 1967 (Cirripedia, Thoracica) and a comparison with other barnacle larvae

The morphological features of the cephalic shield, labrum, abdominalprocess, antennules, antennae and mandibles of Balanus reticulatusare described and illustrated. The size and setation formulaeof the larvae are given at each stage. The trilobed labrum andlateral margin of the cephalic shield with numerous small spinesare diagnostic features for all the subsequent nauplius stages.Numerous small denticulate processes on the surface of the cypridcarapace are major morphological characteristics not found inother balanomorph species. We have constructed keys from stageII to stage VI for the predominant barnacle nauplii of Koreancoastal waters, based on morphological traits such as totallength, shield width, labrum shape, the presence or absenceof posterior shield spines and dorsal shield spines in stagesIV, V and VI, the specific setal type in the fourth segmentof the antennal endopodite, and setation formulae of Pollicipesmitella, Chthamalus challengeri, Megabalanus rosa, B.reticulatus,Balanus amphitrite and Balanus albicostatus.     

Histology and ultrastructure of the salivary glands in Bulla striata (Mollusca, Opisthobranchia)

Abstract. The ribbon‐shaped salivary glands in Bulla striata were studied with light microscopy and transmission electron microscopy (TEM). Secretion is produced in tubules formed by two types of secretory cells, namely granular mucocytes and vacuolated cells, intercalated with ciliated cells. A central longitudinal duct lined by the same cell types collects the secretion and conducts it to the buccal cavity. In granular mucocytes, the nucleus is usually central and the secretory vesicles contain oval‐shaped granular masses attached to the vesicle membrane. Glycogen granules can be very abundant, filling the space around the secretory vesicles. These cells are strongly stained by PAS reaction for polysaccharides. Their secretory vesicles are also stained by Alcian blue, revealing acidic mucopolysaccharides, and the tetrazonium reaction detects proteins in minute spots at the edge of the vesicles, corresponding to the granular masses observed in TEM. Colloidal iron staining for acidic mucopolysaccharides in TEM reveals iron particles in the electron‐lucent region of the vesicles, while the granular masses are free of particles. In vacuolated cells, which are thinner and less abundant than the granular mucocytes, the nucleus is basal and the cytoplasm contains large electron‐lucent vesicles. These vesicles are very weakly colored by light microscopy techniques, but colloidal iron particles could be observed within them. The golf tee‐shaped ciliated cells contain some electron‐dense lysosomes in the apical region. In these cells, the elongated nucleus is subapically located, and bundles of microfibrils are common in the slender cytoplasmic stalk that reaches the basal lamina. The morphological, histochemical, and cytochemical data showed some similarities between salivary glands in B. striata and Aplysia depilans. These similarities could reflect the phylogenetic relationship between cephalaspidean and anaspidean opisthobranchs or result from a convergent adaptation to an identical herbivorous diet.     

Systematic revision of the living species of Bullidae (Mollusca: Gastropoda: Cephalaspidea), with a molecular phylogenetic analysis

Bullidae are a worldwide family of marine shelled cephalaspidean gastropods with a mainly tropical distribution, but also with some representatives in temperate waters. The taxonomy of the group has in the past been based only on shell characters, and the few anatomical accounts available have not addressed more than one to three species, so there has been no agreement about the number of valid species. Seventy‐two specific names and 16 varietal names have been proposed worldwide. The systematics of the family Bullidae are revised, based not only on shells but also on anatomy of all extant species and on DNA sequence data. Twelve species are recognized worldwide, including one new species here described, and all are assigned to the genus Bulla. Two species occur in the eastern Atlantic, B. striata and B. mabillei; two in the western Atlantic, B. occidentalis and B. solida; two in the eastern Pacific, B. gouldiana and B. punctulata; and six in the Indo‐West Pacific, B. ampulla, B. arabica sp. nov. , B. orientalis, B. peasiana, B. quoyii and B. vernicosa. Full synonymies and taxonomic histories are provided for each species. In order to promote taxonomic stability, neotypes are designated for B. striata, B. solida, B. nebulosa (valid name B. gouldiana) and B. vernicosa, and lectotypes for B. occidentalis, B. mabillei, B. punctulata, B. ampulla and B. quoyii. The type locality of B. ampulla is restricted to Mauritius. Bullidae show a general morphological stasis, with anatomy being very similar between species. However, there are high levels of intraspecific variability in the shell, radula and male genital system. In some cases species could only be separated based on molecular data . After defining the characters and geographical range of each species it became clear that sympatric species (a maximum of three) show distinctive shells and reproductive structures, which makes identification straightforward. This study employs an integrative approach, combining information on shells, anatomy, DNA and geographical distribution, in order to resolve the systematics of a difficult taxonomic group. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 153 , 453–543.     

The central nervous system of Bulla gouldiana: Peptide localization

In the present study, we describe the structure of the central nervous system (CNS) of the marine gastropod Bulla gouldiana, and compare it with the structure of the CNS of the related mollusc, Aplysia californica. In addition, we performed an immunohistochemical analysis of a series of peptides, and the synaptic vesicle protein, synapsin I, in the central nervous system of B. gouldiana. The most common peptide in the B. gouldiana nervous system is the molluscan cardioexcitatory peptide (FMRFamide), which is present in a significant proportion of B. gouldiana neurons. A smaller number of neurons exhibit immunoreactivity to antisera raised against the calcitonin gene related peptide, vasopressin, vasoactive intestinal peptide, cholecystokinin, galanin and enkephalin. In some instances there is colocalization of two or more peptides. Very few neurons or axons exhibit synapsin I-like immunoreactivity. The patterns of immunoreactivity to these antisera is quite similar to the patterns that have been described in other gastropods, including Lymnaea stagnalis and Aplysia californica. These observations emphasize the importance of FMRFamide-like compounds in phylogenetically old nervous systems and indicate that compounds similar to mammalian peptides are present in the gastropod. Thus, the production of a wide variety of peptide molecules and their use in neuronal function appears to be a highly conserved phylogenetic process.     

Ontogeny and distribution of Hygophum benoiti(Pisces, Myctophidae) of the North Western Mediterranean

Two species of the lanternfish genus Hygophum, H.benoiti andH.hygomii, are present in the Mediterranean. Differentiationof the larvae from these two species has been problematic. Anattempt to identify such larvae in the plankton samples of theNorth Western Mediterranean led to a broader study that includedthe examination of a large number of plankton samples collectedin the region since 1976. The large number of Hygophum larvaeobtained during July 1992 in the North Western Mediterranean(that proved to be H.benoiti), together with collections oflarvae from different geographic areas (especially H.hygomiifrom the Southwest Indian Ocean), allowed a detailed morphometriccomparison of the larvae of H.benoiti and H.hygomii. Developmentalstages of H.benoiti from yolk sac to post-flexion larvae aredescribed. Information on the temporal occurrence of H.benoitiin the plankton, as well as the spatial distribution of thisspecies during the survey of peak abundance (June/July 1992)in the North Western Mediterranean, is presented.     

The copulatory apparatus of the marine gastropod Haminella solitaria (Heterobranchia: Cephalaspidea) and its phylogenetic relevance

The hermaphroditic marine snail species Haminella solitaria was formerly included in the genus Haminoea, but it was recently assigned to the genus Haminella. The copulatory apparatus in H. solitaria was investigated by light and transmission electron microscopy to obtain additional information about this apparatus in cephalaspidean gastropods and to evaluate the taxonomic relevance of its morphofunctional features in the framework of a new phylogenetic tree of the family Haminoeidae. The copulatory apparatus in H. solitaria consisted of the atrium with a muscular wall and papilla, a seminal duct, and a single‐lobed prostate. Epithelial and subepithelial secretory cells were detected in the proximal and middle region of the atrium wall, and a third type of secretory cell occurred in the distal region of the muscular papilla. The seminal duct was lined by ciliated cells and its muscular wall included some vacuolar cells. The prostate in H. solitaria consisted of lateral pouches surrounding a large central lumen that was filled with spermatozoa. A single type of secretory cell intermingled with ciliated cells formed the epithelium of the prostate. A histological comparison between the copulatory apparatus in H. solitaria and Haminoea navicula revealed substantial differences that support the placement of these two species in different genera, as established by recent molecular studies.     

THE GENUS HAMINOEA (GASTROPODA: CEPHALASPIDEA) IN PORTUGAL, WITH A REVIEW OF THE EUROPEAN SPECIES

The study of specimens of Haminoea collected on the coast ofPortugal, together with the re-evaluation of historical specimensused by Nobre (1938–40) in his main work on the Portuguesemolluscs, recognized the presence of two species, Haminoea navicula(da Costa, 1778) and H. orbignyana (Férussac, 1822),and confirmed the misidentification of H. orbignyana as H. elegans(Gray, 1825). Anatomical analysis found intraspecific variabilityin the radula of H. navicula and H. orbignyana and in the seminalduct of the latter. The systematic value of taxonomic featuresincluding the shell, external morphology, animal coloration,radula, jaws, gizzard plates, prostate gland, seminal duct,penis and egg-masses is discussed. For all European speciesbut H. orbignyana, identification is only possible when anatomicaldata are considered, and the most relevant character is themale reproductive system. SEM images of the penis and prostateof H. navicula and H. orbignyana are given for the first time.A synoptic table for the recognition of the European speciesis provided. (Received 21 January 2005; accepted 17 June 2005)     

Phyllody Malformation in Flowers of Rosaxhybrida cv. Motrea: Effects of Rootstocks, Flower Position, Growth Regulators and Season

Phyllody, a common malformation of Rosaxhybrida cv. Motrea,is characterized by the formation of vegetative structures inplace of stamens and carpels. The incidence of phyllody in thiscultivar is dependent on the type of the root system. The highestproportion of malformed flowers was found in ungrafted plants,this decreased in plants grafted on R. indica major and waslowest on R. canina inermis. Flowers formed on shoots, whichwere located closer to the roots, were more prone to phyllodythan those grown on the upper part of the plants. The incidenceof phyllody was lower in winter than in summer, while shadingof the plants reduced the proportion of malformed flowers regardlessof their root system and the season. Application of gibberellicacid (GA₃) or benzylaminopurine (BAP) greatly promoted the incidenceof phyllody, while paclobutrazol (PBZ) reduced it. The activityof endogenous cytokinins in the xylem exudates from ungraftedplants was higher than in exudates from those grafted on R.indica major, and the lowest activity was found in exudatesfrom plants grafted on R. canina inermis. The involvement ofplant hormones in the development of floral organs in rose flowersis discussed. Key words: Cytokinins, flower malformation, gibberellic acid, homeosis, phyllody, roses     

Casparian Strips in the Leaf Intrastelar Canals of Isoetes duriei Bory, a Mediterranean Terrestrial Species

Anatomical observations, using light and fluorescence microscopy,were made on leaves of Isoetes duriei to verify the presenceof casparian bands around the intrastelar canals. This peculiaranatomical feature, previously reported for some Isoetes species,is confirmed. The possible role of this endodermis-like structure,in the transport of water due to root pressure or in the accumulationof certain metabolites, is discussed in relation to the ecologicaland anatomical features of the species. Copyright 2000 Annalsof Botany Company Isoetes duriei, Lycopsida, Mediterranean, leaf anatomy, endodermis, casparian strips, fluorescence microscopy     

Ecological investigations on the zooplankton community of Balsfjorden, northern Norway: population dynamics of the euphausiids Thysanoessa inermis (Kroyer), Thysanoessa raschii (M.Sars) and Meganyctiphanes norvegica (M.Sars) in 1976 and 1977

The population dynamics of the euphausiids Thysanoessa inermis(Kröyer), T. raschii (M.Sars), and Meganyctiphanes norvegica(M.Sars) have been followed in Balsfjorden in 1976 and 1977.Seasonal variations in length-frequency distributions, growthin carapace length, sex-ratio, and spermatophore productionand attachment are presented and discussed in relation to changesin hydrography and phytoplanlcton standing-crop. An annual generationof T. inermis and T. raschii was spawned in April and May. Eggsand larvae of M. norvegica were not found in Balsfjorden, indicatingthat recruitment occurs from outside the fjord. T. inermis andT. raschii reached maximum carapace lengths of 7–8 mmand 6–7 mm respectively and had life-spans of c. 2 years3 months. M. norvegica had a life-span of c. 2 years 6 monthsand reached a maximum carapace length of c. 12.5 mm. In bothT. inermis and T. raschii 0-group underwent the greatest lengthincreases from May to October, I-group from March to Augustand II-group from April to June. The population structure, growthpatterns and growth periods of M. norvegica were difficult todiscern. The phytoplanlcton cycle appears to be the dominantfactor regulating both growth and spawning of the Thysanoessaspp in Balsfjorden, while temperature has no obvious influence.     

Evidence that the Central Pattern Generator for Swimming in Tritonia Arose from a Non-Rhythmic Neuromodulatory Arousal System: Implications for the Evolution of Specialized Behavior

Comparisons of the nervous systems of closely related invertebratespecies show that identified neurons tend to be highly conservedeven though the behaviors in which they participate vary. Allopisthobranch molluscs examined have a similar set of serotonin-immunoreactiveneurons located medially in the cerebral ganglion. In a smallnumber of species, these neurons have been physiologically andmorphologically identified. In the nudibranch, Tritonia diomedea,three of the neurons (the dorsal swim interneurons, DSIs) havebeen shown to be members of the central pattern generator (CPG)underlying dorsal/ventral swimming. The DSIs act as intrinsicneuromodulators, altering cellular and synaptic properties withinthe swim CPG circuit. Putative homologues of the DSIs have beenidentified in a number of other opisthobranchs. In the notaspid,Pleurobranchaea californica, the apparent DSI homologues (As1–3)play a similar role in the escape swim and they also have widespreadactions on other systems such as feeding and ciliary locomotion.In the gymnosomatid, Clione limacina, the presumed homologousneurons (Cr-SP) are not part of the swimming pattern generator,which is located in the pedal ganglia, but act as extrinsicmodulators, responding to noxious stimuli and increasing thefrequency of the swim motor program. Putative homologous neuronsare also present in non-swimming species such as the anaspid,Aplysia californica, where at least one of the cerebral serotonergicneurons, CC3 (CB-1), evokes neuromodulatory actions in responseto noxious stimuli. Thus, the CPG circuit in Tritonia appearsto have evolved from the interconnections of neurons that arecommon to other opisthobranchs where they participate in arousalto noxious stimuli but are not rhythmically active.     

Composition and seasonality of nocturnal peracarid zooplankton from coastal New Hampshire (USA) waters, 1978-1980

Seasonal changes in the nocturnal peracarid zooplankton froma nearshore area of the Gulf of Maine were studied from January1978 through December 1980 as a requirement of an ecologicalmonitoring program for the Public Service Company of New Hampshire'sSeabrook Generating Stanon. Numerically dominant species includedthe mysids Michteimysis mixta and Neomysis americana. the amphipodsPontogeneia inermis, Themisto gaudichaudi and Monoculodes edwardsi.and the cumaceans Diastylis polita and Lamprops quadriplicata.A general seasonal cycle was documented with a more abundantand speciose assemblage during fall and winter than summer.Peak abundance occurred during spring months prior to emigrationof juvenile M.mixta from the study area. Abiotic and bioticfactors which may affect the structure and composition of thisassemblage are discussed.     

The Suspensor Haustoria of Some Species of Crotalaria Linn

The structure and function of the suspensor has been studiedin eight species of Crotalaria, in all of which it shows a differentiationinto an upper and a lower part. The peripheral cells of thelower part project out and act as haustorial structures forthe absorption of nutrition from the surrounding endosperm.In G. retusa and G.striata these processes are very long andtubular attacking the surrounding endosperm and nucellar tissuesand extending even into the integumental layers. Remains ofthe suspensor are recognizable even in the mature seed. Thecells of the suspensor possess chloroplasts so that this organalso has a photosynthetic function in addition to its haustorialactivity.     

Small-scale distribution of macroplankton and micronekton in the Ligurian Sea (Mediterranean Sea) as observed from the manned submersible Cyana

A series of eight submersible dives (the MIGRAGEL I cruise)was made during late April 1986 using the French submersibleCyana to investigate macrozooptankton in the upper 400–700m of the water column. Paired day and night dives were madeat stations 3, 6, 13 and 23 nautical miles off Cape Ferrat,near Villefranche-sur-Mer, France; the distances represent differentareas in the frontal system of the Ligurian Sea. Detailed day/nightvertical distribution data are shown for the most abundant species;these include the narcomedusa Solmissus albescens, teleost fishCyclothone spp., small appendiculanans (primarily Oikopleuraalbicans), large appendicularians (an undescribed oikopleurid),diphyid siphonophores (mostly Chelophyes appendiculata) andan abundant lobate ctenophore. Salps, pyrosomes, amphipods (Phroniumasedentaria), pteropods (Cavolinia inflexa), macroscopic ‘star-like’protozoa and marine snow are also briefly discussed. The coastalzone was dominated by small appendicularians in the upper layers,with other filter feeders including large appendicularians indeeper water—these just above a non-migratory populationof carnivorous Cyclothone. The carnivorous medusa Solmissusalbescens moved throughout the upper 600 m in the course ofits diel vertical migration. Offshore, carnivores were dominantthroughout the water column, with numerous diphyid siphonophoresin the upper layers, and Cyclothone, lobate ctenophores andmacroprotozoa abundant in deeper water. Solmissus was also present,and was more numerous offshore than in the coastal zone.     

New insights into the functional morphology of the male copulatory apparatus of bullid gastropods

In this study, semi-thin sections stained with histochemical techniques and transmission electron microscopy were used to obtain new data about the morphology and function of the male copulatory apparatus of the cephalaspidean gastropod Bulla striata. The apparatus comprises a vestibule, a penial papilla and a prostate consisting of a coiled unbranched tube ending in a blind caecum. The penial papilla and the coiled tubular prostate are enclosed by a muscular sheath, which is continuous with the muscular tissue of the vestibule. The epithelium lining the lumen of the vestibule is formed by ciliated and mucus-secreting cells. Two new types of subepithelial secretory cells were discovered in this region. The penial papilla is a muscular structure without secretory cells in the epithelium lining the narrow lumen. The tubule that constitutes the prostate possesses a muscular wall and can be divided in three distinct regions: a non-secretory duct connected to the penial papilla, a glandular region rich in large secretory cells and the terminal caecum containing just a few small secretory cells. In the terminal blind caecum, the muscular sheath is fused with the muscular wall of the tubular prostate. Large numbers of spermatozoa were found in the glandular region and in the terminal caecum of the prostate. A new functional mechanism is proposed to explain penial eversion during copulation. This differs from a previous hypothesis in two main aspects: (1) existence of a permanent penial papilla in mature animals acting as a functional penis and (2) functional role of vestibule during copulation, which everts and surrounds the penial papilla, while the latter protrudes outwards.     

PHYLOGENETIC ANALYSIS OF LEARNING-RELATED NEUROMODULATION IN MOLLUSCAN MECHANOSENSORY NEURONS

In spite of significant advances in our understanding of mechanisms of learning and memory in a variety of organisms, little is known about how such mechanisms evolve. Even mechanisms of simple forms of learning, such as habituation and sensitization, have not been studied phylogenetically. Here we begin an evolutionary analysis of learning-related neuromodulation in species related to the well-studied opisthobranch gastropod, Aplysia californica. In Aplysia, increased spike duration and excitability in mechanosensory neurons contribute to several forms of learning-related changes to defensive withdrawal reflexes. The modulatory transmitter serotonin (5-hydroxytryptamine, or 5-HT), is thought to play a critical role in producing these firing property changes. In the present study, we tested mechanosensory homologs of the tail-withdrawal reflex in species related to Aplysia for 5-HT-mediated increases in spike duration and excitability. Criteria used to identify homologous tail-sensory neurons included position, relative size, resting electrical properties, expression of a sensory neuron-specific protein, neuroanatomy, and receptive field. The four ingroup species studied (Aplysia californica, Dolabella auricularia, Bursatella leachii, and Dolabrifera dolabrifera) belong to two clades (two species each) within the family Aplysiidae. In the first clade (Aplysia/Dolabella), we found that the tail-sensory neurons of A. californica and tail-sensory homologs of a closely related species, D. auricularia, responded to bath-applied serotonin in essentially similar fashion: significant increases in spike duration as well as excitability. In the other clade (Dolabrifera/Bursatella), more distantly related to Aplysia, one species (B. leachii) showed spike broadening and increased excitability. However, the other species (D. dolabrifera) showed neither spike broadening nor increased excitability. The firing properties of tail-sensory homologs of D. dolabrifera were insensitive to 5-HT over a wide range of concentrations. We also performed experiments on two outgroup species (Akera bullata and Bulla gouldiana) and found that spike duration was unaffected by 5-HT, whereas excitability was increased. This study suggests that 5-HT-induced spike broadening arose more recently in opisthobranch evolution, whereas 5-HT-induced excitability increase is a more ancestral trait that may have been expressed in the earliest opisthobranchs. Both traits are absent in the aplysiid species D. dolabrifera, demonstrating that a lineage can lose learning-related mechanisms. The phylogenetic variation observed in the present study presents the opportunity to test general models about learning mechanisms and their evolution in unique ways.