Electron microscope studies of spermatogenesis in Branchiobdella pentodonta whitman (Annelida,oligochaeta)

In Brallchiobdella pentodonta Whitman meiosis begins in follicles containing 16 spermatogonia. In each follicle the spermatogonia are connected by cytoplasmic bridges to a central anuclear cytoplasmic mass or cytophorus. They develop synchronously. Synaptonemal complexes are present in the primary spermatocytes. Spermatids contain a large globoid paranuclear body consisting of an acrosomal granule and coiled tubules which evidently receive the contents of the acrosomal granule and are considered the acrosome carrier. The spermatids separate from the cytophorus only when differentiation is completed. The ripe spermatozoon is relatively long. It has anteriorly the coiled tubules, followed by the nucleus, the mitochondrial sphere and the distal centriole from which the flagellum originates, A coiled ribbon-like structure encloses the flagellum along its entire length while a manchette of microtubules surrounds all the other structures of the sperm.     

Ultrastructural aspects of mechano- and chemoreceptors in Branchiobdella pentodonta (annelida,oligochaeta)

The surface receptors in Branchiobdella pentodonta consist of “sense buttons” prevalent on the prostomium, isolated sense cells all along the body of the animal, and free nerve endings. The “sense buttons” are uni- and multiciliated neurosensitive elements and supporting cells together with mucus glandular processes and muscle fibers. In the neurosensitive elements the cilia are always surrounded by cytoplasmic extroversion. The cytoplasm of the apical zone has abundant small dense granules, mitochondria, bands of tonofilaments, and microtubules. The cilium of uniciliated elements originates from three short roots. The highly vacuolated support cells surround the neurosensitive elements, separating them from each other. The “sense buttons” appear to be mechanoreceptors and chemoreceptors, and the isolated sense cells tactile mechanoreceptors, as are the free nerve endings. The surface receptors are compared with those of other Oligochaeta and Hirudinea.     

Ultrastructure of the mouth apparatus in Branchiobdella pentodonta (annelida,clitellata)

The mouth apparatus in Branchiobdella pentodonta was studied by electron microscopy. The opening is situated dorsal to the adhesive disk of the anterior sucker and is surrounded by a ring of 16 papillae. The papillae have mono-layered epithelium, muscle fibers, glandular processes, and taste and olfactory organs like “sensitive buttons.” The oral cavity contains jaws with horny teeth and “sensitive buttons,” and is surrounded by a ring of circular muscle fibers that connect to the muscle fiber of the papillae. This apparatus shows some analogies to that of Hirudinea, such as the presence of sucker and jaws with horny teeth.     

Immunohistochemical localization of bioactive substances in the epidermis of the earthworm, Lumbricus terrestris (Annelida, oligochaeta)

Serotonin (5HT), endothelin-big (ET-big) and neuron-specific enolase (NSE) are present in some epithelial paraneuronal cells of vertebrates. In the invertebrates, we show for the first time, by immunofluorescence and immunoperoxidase methods, the immunoreactions to antibodies raised to ET, 5-HT and NSE in the epidermis of the Lumbricus terrestris. It is assumed that the reactive cells are small granular cells that may have a paracrine action in the mechanisms of cell proliferation and secretory processes. The function of these bioactive substances in the epidermis of the earthworm awaits investigation. Phylogenetically it is very interesting that paraneuronal-like type of cells appear in such an ancient species of invertebrate which uses the epidermis for various functional activities.     

Morphological reinvestigation and phylogenetic relationship of Acanthobdella peledina (Annelida,Clitellata)

Summary In recently collected specimens of Acanthobdella peledina the nervous system, the genital organs and the coelomic organisation were reinvestigated after complete serial sections. These anatomical results are schematically represented. In addition, the integument, the chaetae and the peripheral muscle layer were investigated by electron microscopy. In general, the results confirm Livanow's classic monograph (1906), with the exception of a few details. The body apparently possesses neither a prostomium nor an achaetous buccal region (peristomium). The number of 29 true segments is concluded from the number of segmental ganglia. The five anteriormost segments, each with four pairs of hookshaped chaetae arranged around the mouth opening, are considered to be functionally equivalent to an anterior sucker. The ultrastructure of the integument and the chaetae generally conforms to the typical annelidan pattern. The muscle cells are of the typical hirudinean type. The outer male genital pore is positioned in segment 10; the female organs open in segment 11 directly behind the septum between segments 10 and 11. The main emphasis is laid on the evaluation of the position of the taxon within the Clitellata, including a discussion of the Branchiobdellida, and the cladograms presented show the Acanthobdellida to be the sister group of the Euhirudinea. Characters shared by the Branchiobdellida and Hirudinea (including A. peledina) are considered to be convergently evolved.     

Comparative electron microscopic studies of muscle and sperm cells inBranchiobdella pentodonta whitman andBythonomus lemani grube (Annelida clitellata)

Summary To determine the degree of systematic affinity between branchiobdellids and lumbriculids, the ultrastructures of muscle and sperm cells ofBranchiobdella pentodonta andBythonomus lemani were studied.The structure ofBranchiobdella sperms is similar to that of the oligochaete model in terms of nucleus and the middle piece and to that of the hirudinean model in the peculiarity of the flagellum and acrosomal complex. Muscle ofBranchiobdella are very similar to those of the hirudineans.Muscles and sperms ofBythonomus lemani are practically identical to those of the tubificid model and are closely related to the other known oligochaetes.This paper is dedicated to Professor Silvio Ranzi to celebrate his 75th birthday     

New aspects of the possible sites of ultrafiltration in annelids (oligochaeta)

Electron microscopic investigations of blood vessels were conducted to show sites of filtration such as podocytes or fenestrated endothelia. The endothelia of the blood vessels of Aelosoma hemprichi, Nais elinguis, Dero obtusa and Enchytraeus buchholzi consist of myoendothelial cells, chloragocytes and podocytes. The podocytes form large archs over a considerable area of the vessels. On the lumen side of the vessel there are several columnar processes which split into numerous small pedicels. The gaps between the adjacent pedicles are bridged by slit membranes. The podocytes are restricted to the front part of the ventral vessel. They are presumed to form a filtration surface. Furthermore, some parts of the ventral vessel are formed by a fenestrated endothelium, mainly in Enchytraeus buchholzi. In the vascular system of E. buchholzi two separate filtration sites were found. Additionally to the filtration site between ventral vessel and coelomic cavity a second filtration site was found in the front part of the body between blood sinus and coelomic cavity. In such areas the basement membrane is the only continuous layer between the blood vessel and the coelomic cavity. Its thickness is in the range of 40 nm. Possible filtration sites in the form of podocytes and irregular fenestrations could be localized at the border between the blood compartment and the coelomic compartment. It can be presumed that the primary urine may be formed by ultrafiltration of blood.     

Morphological variation in Polydora ciliata complex (Polychaeta: Annelida)

Intraspecific variation in Polydora ciliata (Johnston) was assessed on the basis of evidence from morphological characters, which were studied by scanning electron microscopy. Differences were observed between populations of P. ciliata with respect to the setae of the fifth modified segment and in the caruncle. Two other species, namely P. ligni Webster and P. limicola (Annenkova), were also studied in order to assess interspecific variation. The two forms of P. ciliata , that is boring and non-boring, were found to be different morphologically.     

The skin of Osedax (Siboglinidae,Annelida): An ultrastructural investigation of its epidermis

The symbiotic polychaetes of the genus Osedax living on the bones of whale carcasses have become known as bone‐eating worms. It is believed that whale bones are the source of nutrition for those gutless worms and that fatty acids are produced by their symbionts and transferred to the host. However, the symbionts are of the heterotrophic group Oceanospirillales and as such are not able to synthesize organic carbon de novo. Also, they are not housed in close contact to the bone material. We studied the ultrastructure of the integument overlying the symbiont housing trophosome in the ovisac region and the roots region and of the symbiont‐free trunk region of Osedax to investigate the host's possible contribution in feeding for the whole symbiosis. The epidermis differs conspicuously between the three regions investigated and clearly points to being correlated with different functions carried out by those regions. The ultrastructure of the integument of the root region changed towards the ovisac region and corresponds with the change of the ultrastructure observed in the Osedax trophosome. We suggest that the epidermis in the root region is tightly linked to bone degradation and nutrient uptake. The trunk region possess two types of unicellular gland cells, at least one of which seems to be involved in secretion of the gelatinous tube of adult Osedax females. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

An ultrastructural study of spermiatogenesis and spermatozoal morphology in Sparganophilus tamesis (Sparganophilidae, Oligochaeta, Annelida)

Spermatogenesis in Sparganophilus tamesis results in a filiform spermatozoon of the advanced type. Structures in proximal-distal sequence are the slender acrosome, the extremely elongate nucleus, the short cylindrical midpiece and the long flagellum. The acrosome is unique in those Lumbricina so far studied in persistence of the subvesicular space in the anterior (proximal) bulb formed by the primary acrosome vesicle, external to the acrosome tube and in extension of the acrosome rod into this proximal subvesicular space. A further unusual feature is the very long basal chamber in the acrosome tube, distal to the rod, as in the Phreodrilidae. The acrosome tube has a distal limen and its junction with the nucleus is domed. There is little similarity with the acrosome of the Lumbricidae, a family with which the Sparganophilidae have recently been grouped. The midpiece has the highest number of mitochondria yet recorded for oligochaetes. The number is deduced to be eight, of which seven are normally visible in cross section because one of the mitochondrial loci consists, uniquely for the Oligochaeta, of two mitochondria aligned longitudinally in tandem. The structure of the flagellum, including the arrangement of its peripheral glycogen granules, is typical for oligochaetes. The spermatozoon is straight, lacking a spiral coil or keel. These results are discussed in relation to oligochaete phylogeny.     

The cocoon-producing cells of Eisenia foetida (Annelida, Oligochaeta): a histochemical and ultrastructural study

Type 1 cells of the clitellar epithelium of Eisenia foetida secrete a protein resembling keratin in histochemical reaction. Ultrastructurally, type 1 cells are characterized by membrane-bound, pockmarked granules ranging in diameter from 1.0 to 3.0 micrometers. Immature granules often exhibit an organized microfibrillar substructure. Individual microfibrils are 17 +/- 1 nm in diameter. Type 2 cell secretion contains protein, nonsulfated acid mucosubstance, and neutral mucosubstance with 1,2-glycol groups. The coarsely fibrillar granules are membrane bound and vary in diameter from 0.5 to 1.0 micrometer. The necks of both type 1 and type 2 cells contain a peripheral ring of microtubules, 20 +/- 1 nm in diameter.     

Morphological and molecular evidence of the phylogeny of Nereidiform polychaetes (Annelida)

The phylogeny of Nereidiformia is assessed in a parsimony analysis of combined morphological and DNA data, with special focus on previously questioned relationships between Chrysopetalidae and Hesionidae, between Pilargidae and Hesionidae, and the affinities of Hesionides and Microphthalmus. A 660 bp segment of the mtDNA cytochrome c oxidase subunit I gene was sequenced for two chrysopetalids, one nereidid, one pilargid, one pisionid, two hesionids, plus the two questionable hesionids Hesionides arenaria and Microphthalmus sp. Phylogenetic resolution was poor for the cytochrome c oxidase subunit I gene data alone, but the combined analysis yielded partially robust topologies, suggesting that nereids are the sister group to chrysopetalids, and that pilargids, Hesionides and Microphthalmus do not belong within the hesionids     

Stigmatic and stylar incompatibility in Prunus avium L.: Morphological and ultrastructural aspects

Abstract

Stigmatic and stylar structures of-sweet cherry (Prunus avium L.) «Malizia» were examined with light and electron microscopes (S.E.M. and T.E.M.).

It was demonstrated that the transmitting tissue, situated in the central portion of the style, extends below the stigmatic papillae and secretes an electrondense material accumulating in the intercellular spaces. Pollen tubes which germinated on the stigma, reached the ovules passing through this substance which facilitates their passage while trophic relations are established.

Some ultrastructural aspects of the pollen tubes observed inside the style were examined and discussed in relation to the phenomenon of self-incompatibility.     

Morphological comparison of the regeneration process in Sabella spallanzanii and Branchiomma luctuosum (Annelida,Sabellida)

Within Annelida, the family Sabellidae is a particularly interesting clade in which to carry out comparative studies of regeneration, as its members vary in regeneration potential, with some even lacking this ability completely. However, data on the regenerative potential of sabellids are available only for a few species investigated early in the last century, and even for those, many aspects of regenerative ability are yet to be determined. In the present study, we compared the morphology of the regeneration process of anterior and posterior ends in Sabella spallanzanii and Branchiomma luctuosum, two sabellids that belong to closely related genera. Members of S. spallanzanii exhibited lower mortality in response to wounding and greater regenerative ability, with amputees able to complete the regeneration process more rapidly than in B. luctuosum. Moreover, results from this study suggest that adults of S. spallanzanii and B. luctuosum use different mechanisms to restore lost anterior body parts. In S. spallanzanii, both morphallaxis and epimorphosis appear to be important in anterior end regeneration, but in B. luctuosum, only epimorphosis appears to be important. Such variation among closely related taxa sharing similar body plans has rarely been reported, but might be profitably examined using molecular approaches in these and other species of sabellids. Results of such studies interpreted in the context of phylogeny may ultimately provide insight into the evolution of regeneration.     

Immunohistochemical (cLSM) and ultrastructural analysis of the central nervous system and sense organs in Aeolosoma hemprichi (Annelida, Aeolosomatidae)

The Aeolosomatidae are very small limnetic or terrestrial annelids of apparently simple organisation and uncertain phylogenetic position. They have been placed either at the base of the Clitellata, as a highly derived taxon within the Clitellata closely related to the Naididae, or as their sister group within the „Polychaeta”. A combined immunohistochemical (cLSM) and ultrastructural investigation of the central nervous system and the sense organs in Aeolosoma hemprichi was undertaken to look for characters which might support one of these theories. The position of the brain within the prostomium and the organisation of the ventral nerve cord, with its intraepithelial paired longitudinal nerves lying far apart from each other and the presence of a median longitudinal nerve, are atypical for the Clitellata and clearly differ from the situation found in Naididae. Moreover, the circumoesophageal connectives are bifurcated and enter the brain as dorsal and ventral roots; this arrangement is unknown in Clitellata, in which these connectives are unbranched. An ultrastructural analysis of the ciliated pits located laterally in the furrow between prostomium and peristomium in A. hemprichi and other Aeolosomatidae show that they are in fact nuchal organs. Such presumed chemosensory organs are typical of „Polychaeta” and absent in all Clitellata. Two pairs of ciliary sense organs are present in the prostomium in front of the brain of A. hemprichi. Although similarly organised sensory structures occur in many species of the Clitellata, they differ in position and certain ultrastructural features and are known from other Annelida as well. These results clearly support the exclusion of the Aeolosomatidae from the Clitellata and do not provide any evidence for a sister-group relationship between these two taxa. Accepted: 8 February 2000     

Morphological, palaeontological and molecular aspects of ichneumonoid phylogeny (Hymenoptera, Insecta)

Ichneumonoid phylogeny is revised on the basis of morphological, palaeontological and molecular evidence. The only previous formal cladistic study of the phylogeny of the families of the superfamily Ichneumonoidea made many assumptions about what families lower taxa belonged to and was based on a very limited set of characters, nearly all of which were uninformative at family level. We have subdivided both Ichneumonidae and Braconidae into major groups, investigated several new character systems, reinterpreted some characters, scored several character states for extinct taxa by examining impression fossils using environment chamber scanning electron microscopy, and included data for a significant new subfamily of Braconidae from Cretaceous amber of New Jersey. Sixteen different variants of the data set were each subjected to parsimony analysis without weighting and with successive approximations weighting employing both maximum and minimum values of both the retention and rescaled consistency indices. Each analysis resulted in one of seven different strict consensus trees. Consensus trees based on subsets of these trees, selected on the basis of the optimal character compatibility index (OCCI), resulted in an eighth distinct tree. All trees had the Braconidae monophyletic with the Trachypetinae as the basal clade, and also had a clade comprising various arrangements of Apozyginae, the Rhyssalinae group, Aphidiinae and 'other cyclostomes', but relationships among the remaining braconid groups varied between trees. Only one of the consensus trees had the Ichneumonidae (including Tanychorella ) monophyletic. The Eoichneumonidae + Tanychora are the sister group the Braconidae in two of the consensus trees. Paxylommatinae were basal in the clade comprising the Eoichneumonidae + Tanychora and the Braconidae. The preferred tree, based on the highest OCCI was used for interpreting character state transitions.