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The nervous system of the male Dinophilus gyrociliatus (Polychaeta, Dinophilidae): II. Electron microscopical reconstruction of nervous anatomy and effector cells.

All neuronal cells in the dwarf male of the dimorphic polychaete species Dinophilus gyrociliatus were individually identified by means of serial ultrathin sections. Altogether 68 neural cells--including 40 sensory neurons and 2 glial cells--constitute a small but complex nervous system. Fifty-three neural cells are located in three pairs of ganglia and connected by paired nerve cords. The prominent frontal ganglia, each consisting of a well-developed neuropile and surrounded by 20 or 21 neural cells, represent the animal's brain. The ventral ganglia contain only 2 neurons each. The penis ganglia--four cells each--are associated with the copulatory organ. A conspicuous circumpenial fiber mass surrounds the basal part of the penis. The effector cells--22 multiciliated epidermal cells, 34 muscle cells, and different gland cells (?)--were also reconstructed and their innervation was partly elucidated. Sensory-motor neurons were unambiguously identified. They are discussed in regard to the small body size of the animal. The male's nervous organization resembles a very simple rope ladder and may represent a reduced derivative of a nervous system in normal-sized males of monomorphic species. Similarities, however, also occur with the developing nervous system of a planktotrophic metatrochophore. The neuronal organization, with its two centers (frontal ganglia and ventral ganglia vs. penis ganglia and circumpenial fiber mass), accords well with the bipartite behavioral pattern, which is entirely devoted to locomotion and copulation, respectively.

Animals↗

Serotoninlike immunoreactivity in the central and peripheral nervous system of the scale worm Harmothoe imbricata (Polychaeta).

The distribution of serotoninergic neurons in the nervous system of the scale worm Harmothoe imbricata was visualized in the anterior half of the body by the peroxidase-antiperoxidase (PAP) immunohistochemical method with a specific antiserotonin antibody. Immunoreactive neuronal somata were localized in discrete ganglion cell masses of the dorsally situated cerebral ganglion and in segmental ganglia of the ventral nerve cord. They also make up the majority of neurons present in the parapodial ganglia. Large and small varicose fibers stained in the neuropile of all the above-mentioned ganglia but also in interganglionic connectives and segmental nerves. On the basis of soma size and location and of fiber distribution, the reactive neurons were identified as primarily interneuronal with a few motoneurons and presumptive afferent neurons. The presence of a motor component was substantiated by observations of several reactive varicose fibers spread over longitudinal muscle layers of the trunk. In addition, neurites of the subepidermal nerve plexus and enterochromaffinlike cells of the gut epithelium reacted with the serotonin antibody. It is concluded that serotoninergic pathways are ubiquitous elements in the organization of the central and peripheral nervous system of this polychaete. The significance of these findings in relation to other annelid groups and to the physiological role of serotonin is discussed.

Animals↗

Nephridial and gonoduct distribution patterns in Nerillidae (Annelida: Polychaeta) examined by tubulin staining and cLSM.

The distribution and configuration of nephridia and gonoducts are described for seven species from seven genera of the interstitial polychaete family Nerillidae. The ciliated nephridia and gonoducts were identified by tubulin staining and examined with a confocal laser scanning microscope. The following species of the seven to nine-segmented nerillids were examined: Leptonerilla prospera, Nerilla antennata (nine segments); Nerillidium mediterraneum, Trochonerilla mobilis, Gen. sp. A (eight segments); and Aristonerilla brevis, Paranerilla limicola (seven segments). Two of the examined species are hermaphroditic (N. mediterraneum and Gen. sp. A). Segmented nephridia can be found from the first to the last segment, with a total of two to five pairs. One to three pairs of segmented spermioducts are present in all species. One pair of gonoducts is found in all species, except for P. limicola, where they are absent. Nephridia vary in length from half to almost twice the length of a segment and may be curled up in loops. In A. brevis and P. limicola the nephridia are discontinuously ciliated. The distribution and configuration of spermioducts and gonoducts are also variable, although to a lesser extent. The spermioduct distribution is generally consistent within genera and therefore of systematic significance. Nephridia and gonoducts are never found together in the same segments, and the results indicate that gonoducts and nephridia have developed from the same anlagen. The distribution patterns of nephridia and gonoducts are discussed with respect to segmentation, systematics, and development.

Animals↗

Cell proliferation dynamics and morphological differentiation during regeneration in Dorvillea bermudensis (Polychaeta, Dorvilleidae).

Although some species of Annelida have an enormous capacity to regenerate, it is not yet known whether reestablishment of lost body parts is performed by stem cells, depends on preceding dedifferentiation of somatic cells, or is a combination of both. In order to clarify how, in the case of epimorphic regeneration, the blastemas are formed, we applied the thymidine analog 5'-bromo-2'-deoxyuridine (BrdU) in the dorvilleid polychaete Dorvillea bermudensis to identify cells in the S-phase of the cell cycle. Regeneration pulse-chase experiments were carried out to determine onset and dynamics of the proliferation process, and BrdU pulse-chase experiments were undertaken to follow cell fate. We found irregularly distributed S-phase cells throughout the body of adult specimens. Subsequent to amputation, these cells do not migrate from the amputee towards the wound site, where proliferation activity was documented no earlier than 16 h after fragmentation. In the initial phase, the proliferation rate at the anterior end exceeds the rate at the posterior end. Observance of identity could be demonstrated for the ectoderm and can be assumed for the two other germ layers. The anterior blastema transforms into the head, while the posterior forms the pygidium and persists as a proliferation zone; four or numerous segments are formed by intercalation between the former anterior or posterior blastema and the amputee.

Animals↗

CLSM analysis of the phalloidin-stained muscle system in Nerilla antennata, Nerillidium sp. and Trochonerilla mobilis (Polychaeta; Nerillidae).

The entire muscle system of Nerilla antennata, Nerillidium sp. and Trochonerilla mobilis was three-dimensionally reconstructed from whole mounts. In juvenile and adult specimens the F-actin musculature subset was stained with FITC-conjugated phalloidin and visualized with a confocal laser scanning microscope (cLSM). The muscle system shows the following major organization: 1) circular muscles are totally absent in the body wall; 2) the longitudinal muscles are confined in two ventral and two dorsal thick bundles; 3) additional longitudinal muscles are located in the ventro- and dorsomedian axis; 4) three segmental pairs of ventral oblique muscles elongate into the periphery: the main dorsoventral muscles that run along the body side posterior and dorsally and the anterior and posterior oblique parapodial muscles, which contribute to the ventral chaetal sacs; 5) one segmental pair of dorsal oblique parapodial muscles, contributing to the dorsal chaetal sacs; 6) five to seven small dorsoventral muscles per segment; and 7) complex head and pharyngeal musculature. These results support the belief that absence of circular muscles in the polychaete body wall is much more widely distributed than is currently presumed.

Animals↗

Clonal domains in postlarval Platynereis dumerilii (Annelida: Polychaeta).

Following an enzymatic procedure for softening the egg envelope, blastomeres in the embryo of the polychaete Platynereis dumerilii were injected with TRITC-dextran. Injection was successful in the following blastomeres: AB, CD, A, B, C, D, 1a-1d, 1A-1D, 4d, and 4d(1). The distribution of fluorescent label was recorded by confocal laser scanning microscopy of young, three-segmented worms after 3 or 4 days of development, in some cases also in 1-day-old trochophore larvae. Results were documented by single optical sections, by stacking a limited number or a complete set of optical sections, and by computer-generated surface views of both the labeled tissue domains and the body contours from complete image stacks of whole worms. With respect to their descent from the embryonic cell pattern, five major compartments can be distinguished which together compose the body of the young worm: 1) The epispheric, epidermal, and neural region of the head, composed of four domains arranged as quasi-radial sectors derived from micromeres 1a, 1b (left and right ventral), and 1c and 1d (right and left dorsal). 2) A posttrochal epidermal region of the head originating from micromeres 2a(1)-2c(1) and constituting the ventral and lateral posttrochal epidermis of the head. 3) A stomodeal-ectomesodermal region of the head, including the stomodeum (micromeres 2a(2) and 2c(2)), its mesodermal envelope, and head mesoderm (micromeres 3a-3d). 4) A solid cone composed of the four terminal macromeres 4A-4D, forming the core of the trunk as the endoderm anlage. 5) An epidermal and mesodermal coating of the trunk originating from the dorsal micromeres 2d and 4d. The region of the so-called (first, anterior) peristomial cirri at the posterior flanks of the head is also composed of 2d- and 4d-derived trunk tissue, thus corroborating the postulated descent of this region and its appendages from a cephalized anteriormost trunk segment and its parapodia. The cell-lineage domains of the first and third micromere tiers are arranged left or right of the sagittal plane, while two micromeres of the second quartet are in a lateral and, initially, two in a median position (2b ventral and 2d dorsal). The offspring of micromere 2d expand from a dorsal position toward the ventral midline and those of cell 4d from a posterior-dorsal site toward the anterior, initially forming two lateral bands. In the epispheric part of the head, part of the neurectodermal tissue derived from micromeres 1a and 1b interweaves in a medio-sagittal bar, and part of the first micromere offspring of all four quadrants (1a-1d) combine in forming a central brain neuropil. Each of the latter sends neurites through both of the circumesophageal connectives. Paired muscle tracts extend through the head toward the base of the antennae and are probably derived from micromeres 3a and 3b. A mesodermal envelope of the stomodeum is probably built by the 3c and 3d micromeres. The formation of symmetry and the nature of the body axes in the embryo and adult of Platynereis dumerilii are discussed. J. Morphol.

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A PCR-based survey of homeobox genes in Ctenodrilus serratus (Annelida: Polychaeta).

The polychaete worm Ctenodrilus serratus was surveyed for the presence of HOM/HOX and engrailed-type homeobox genes using PCR with degenerate primers. Sixteen unique homeobox fragments were found in surveys of genomic and cDNA with three different primer sets. For three fragments, RACE was employed to obtain additional homeobox sequence and the 3' flanking region. Nine HOM/HOX-type fragments were identified, including putative representatives of the Hox1/lab, Hox2/pb, Hox3, Hox4/Dfd, and Antp/Lox5 cognate groups. Two additional Antp-like fragments could not be assigned specific orthology. Presence of an ortholog of leech Lox2 in addition to a Ubx/abdA-like gene suggests that independent duplications of a single precursor occurred in the annelid and arthropod lineages. No representative of the Hox9/AbdB group was identified. Our results are consistent with a hypothesis of a single HOM/HOX cluster in Ctenodrilus as extensive as that seen in strongly tagmatized arthropods, suggesting that the primitive role of these genes even in overtly metameric animals was something other than specification of overt segmental differentiation. The primers used also detected representatives of six other homeobox classes or families: Xlox (XlHbox8/HTr-A2), Ovx (Chox7), caudal, Prh (proline-rich homeobox), NEC (ceh-9/Tghbox5), and engrailed.

Amino Acid Sequence↗

A light- and electron-microscopic investigation of gametogenesis in Typosyllis pulchra (Berkeley and Berkeley) (Polychaeta: Syllidae). I. Gonad structure and spermatogenesis.

Tryposyllis pulchra reproduces by the production of three to four gamete-bearing stolons (schizogamy) during consecutive 30--day periods. Although gonads are found in a large number of segments, only those in the posterior-most segments produce gametes and become incorporated into the developing stolon. The more anterior gonads remain undifferentiated and probably sexually undetermined until they are needed in future stolonizations. Gonial cells, which will eventually become either male or female, are ultrastructurally identical at the onset of each stolonization period. Spermatogenesis is marked by a short proliferative period followed by differentiation and spermiogenesis. The first ultrastructural signs of spermatogenesis were found in coelomic spermatogonia on day 10 of stolon formation. Spermatogonia are joined by intercellular bridges, which are maintained until the early spermatid stage. Synaptonemal complexes mark the onset of meiosis, which is apparently synchronized in the syncytial clusters of primary spermatocytes. Spermiogenesis occurs during the final 10 days of stonolization and a variety of stages is present within a single animal. All sperm mature by the time the stolon detaches. Acrosome formation and nuclear condensation are described in addition to the ultrastructure of mature sperm.

Acrosome↗

A light- and electron-microscopic investigation of gametogenesis in Typosyllis pulchra. (Berkeley and Berkeley) (Polychaeta: Syllidae). II. Oogenesis.

Oogenesis in Typosyllis pulchra begins with a short proliferative gonial phase. Gonial clusters, consisting of six to eight syncytial sibling cells, are surrounded by gonial peritoneum and are eventually liberated into the coelom. At about the same time as the onset of the coelomic phase, one gonial cell in each cluster differentiates, begins to accumulate yolk and increases in size, eventually to become a mature oocyte. Yolk is composed of lipid droplets and membrane-bound protein granules. The protein component appears to be at least partially synthesized by the oocyte. Yolk is composed of lipid droplets and membrane-bound protein granules. The protein component appears to be at least partially synthesized by the oocyte although there are some signs of endocytosis. Nurse cells and peritoneal cells are apparently able to manufacture some protein that may also be utilized by the oocyte. The nurse cells maintain cytoplasmic continuity with the oocyte, and mitochondria and ribosomes are often seen within the intercellular bridges between oocyte and nurse cell. Immediately prior to spawning, oocytes undergo prematuration and cytoplasmic reorganization. Ultrastructural changes which occur during oocyte growth and maturation are described.

Animals↗

Acrosome formation and the centriolar complex in the spermatozoa of Sabella penicillum (Polychaeta): an electron microscopical study.

The acrosomal vesicle of Sabella penicillum spermatids consists of an electrondense core and a more transparent surrounding zone. During subsequent differentiation the vesicle membrane forms several invaginations in the juxtanuclear area. These invaginations later establish contact with the core. In the mature spermatozoon the spaces between the invaginations appear as electron-dense "tubules"; this is probably due to a shift of material from core to periphery. The ultrastructure of the centriolar complex is described in detail.

Acrosome↗

Discocilia--a new type of kinocilia in the larvae of Lanice conchilega (polychaeta, terebellomorpha).

A modified type of kinocilia has been found in the Aulophora-Iarva of the sedentarian polychaete Lanice conchilega. For this newly described cilium type the term "discocilium" is proposed. The only structural difference from usual locomotory cilia is the tip, which possesses a discoidal head. The head is formed from the terminal part of the cilium shaft, which is bent to give rise to a loop-like ring covered by the ciliary membrane. Three types of discocilia can be distinguished: a) discocilia having swollen, bulblike heads with a central straight axoneme; b) discocilia having heads with a curved lateral axoneme and c) discocilia in which the axoneme forms a loop. The internal structure shows the usual 9 + 2 arrangement of the filaments. The head shows no sign of secretion; it appears structureless in electron microscopical examination. There are two kinds of discocilia arrangements: 1) isolated bunches of cilia especially at the tentacles and in the frontal region, and 2) segmental dorsal rows of cilia. The possible formation of discocilia is described.

Animals↗

Unusual granules in the ejaculatory duct of a Microphthalmus species (Polychaeta, Annelida).

The paired prominent ejaculatory ducts of the hermaphroditic polychaete Microphthalmus cf. listensis are surrounded by gland cells the processes of which penetrate the ducts themselves. These cells produce, in separate regions, two different types of spherical granules. Type I is composed of an electron dense and an electron lucent part. Type II granules contain a tubular filament that forms a single or double spiral in the periphery of a more or less unstructured electron dense material. Golgi vesicles give rise to this granule type. During the passage of sperm, these granules are obviously discharged into the lumen of the duct. Here they change form and probably dissolve. Their function is as yet unknown; capacitation of sperm is assumed.

Animals↗

Autoradiographic detection of indolamine and catecholamine neurons in the nervous system of Owenia fusiformis (Polychaeta, Annelida).

Light- and electron-microscopic autoradiographic studies were carried out on Owenia to detect selectively catecholaminergic and indolaminergic neurons at two appropriate body levels according to the regenerative properties of this annelid, i.e. in the 1st and the 4th abdominal segments. Autoradiographically, intense 3H-5-hydroxytryptamine accumulation is seen in the 1st abdominal segment, and a less intense autoradiographic reaction in the 4th segment. Roughly, a similar difference in the distribution of the label is seen at both levels following 3H-noradrenaline administration. These observations, confirmed by a quantitative uptake analysis, suggest the presence of catecholaminergic and indolaminergic neurons in Owenia.

Animals↗

Cellular immunity in an annelid (Nereis diversicolor, Polychaeta): production of melanin by a subpopulation of granulocytes.

We attempted to identify the nature and origin of the pigment produced by the marine worm Nereis diversicolor in order to isolate, in inert brown capsules, foreign objects introduced into its body cavity. This brown pigment, characterized by cytochemical techniques, could be a melanin. The activity of the enzyme phenoloxidase responsible for melanin biosynthesis was detected by enzyme cytochemistry techniques in vacuoles and the Golgi apparatus of coelomocytes activated by the presence of foreign bodies. Morphological techniques combined with a monoclonal immunological probe enabled us to establish that the "G2" granulocytes contain both the precursor of the pigment in dense bodies and the capacity for phenoloxidase synthesis when activated to encapsulate foreign bodies. The "G2" granulocyte may therefore be compared to a melanocyte in which melanin is not stored as in mammals, but immediately extruded following synthesis in the form of a thick fluid.

Animals↗

Localization of a vertebrate telomeric sequence in the chromosomes of two marine worms (phylum Annelida: class polychaeta).

Using the fluorescence in situ hybridization (FISH) technique, the presence of the vertebrate telomeric sequence (TTAGGG)n was found in the chromosomes of two marine polychaetes belonging to two separate orders: one errant, Platynereis dumerilii (family Nereidae), and the other sessile, Pomatoceros lamarckii (family Serpulidae). This sequence was exclusively present at the ends of the chromosomes in both species.

Animals↗

Comparison of cellular and whole-animal bioassays for estimation of radiation effects in the polychaete worm Neanthes arenaceodentata (Polychaeta).

The polychaete worm Neanthes arenaceodentata was used in experiments to determine possible relationships between short-term genotoxicity tests and reproductive and lethal consequences of exposure to ionizing radiation. Groups of juvenile N. arenaceodentata received one of four different radiation doses (2, 4, 8, and 16 Gy) to determine dose-effect estimates for chromosomal aberration induction, and groups of both adult and juveniles received one of seven different radiation doses (1, 4, 8.4, 46, 102, 500, and 1000 Gy) to determine dose-effect estimates for reproduction, mortality, and life span. Effects on reproduction and genetic material were observed at the lowest doses and in the same range; detrimental reproductive effects were observed at 1 to 4 Gy, and the frequency of chromosomal aberrations was significantly increased at 2 Gy. Only high doses resulted in acute mortality (greater than 500 Gy) and decreased life span (greater than 100 Gy). Dose-effect estimates for chromosomal aberration induction were dependent on radiation dose and on the stage of the cell cycle at the time of irradiation. Dose-effect estimates for reproduction were dependent on dose and the potential for repopulation of gonadal tissue. It is concluded that short-term genotoxicity test can be predictive of detrimental reproductive effects in those model systems for which basic cell kinetics and reproductive parameters are well known.

Animals↗

Petroleum hydrocarbon resistance in the marine worm Neanthes arenaceodentata (polychaeta: annelida), induced by chronic exposure to No. 2 fuel oil.

1. Three successive generations of the marine polychaetous annelid Neanthes arenaceodentata taken from a laboratory population, were continuously exposed to one of three sublethal concentrations of No. 2 Fuel Oil water-soluble-fraction (WSFs). During each generation larvae, juvenile, and immature adult polychaetes were challenged with acute (96 hr) doses of No. 2 Fuel Oil or south Louisiana crude oil WSF to test their sensitivity to petroleum hydrocarbons (PHCs). 2. Larvae from all 3 generations, at all exposure concentrations, were no different from control (susceptible) larvae in their sensitivity to the two test oils. F1, F2, and F3 adults exhibited equally increased PHC resistance (X2) compared to control adults. 3. The only evidence of increased resistance beyond that observed in F1 animals was seen in results of bioassays with juvenile worms, wherein PHC resistance increased from slightly below control levels in F1 juveniles to slightly above control tolerance among F3 juveniles. 4. With the exception of F1 worms, removal from chronic exposure 7 or 14 days prior to challenge did not result in termination or reduction of resistance, implicating a genetic mechanism behind PHC resistance in N. arenaceodentata. 5. F3 resistant and unexposed control polychaetes accumulated, metabolized, and excreted a key diaromatic PHC (naphthalene-14C) in quantitatively identical fashion. Mechanisms responsible for resistance appeared unrelated to external permeability and/or excretion rates.

Animals↗

In vivo measurements of the internal pH of Hediste (Nereis) diversicolor (Annelida, Polychaeta) exposed to ambient sulphidic conditions using pH microelectrodes.

The effect of different ambient sulphide concentrations on the internal pH regime of Hediste (Nereis) diversicolor was studied under in vivo conditions using liquid membrane pH microelectrodes, a method which is new to marine sciences. As a case study, the hypothesis was tested whether organisms exposed to ambient sulphidic conditions are able to lower their internal pH which, in effect, would reduce sulphide influx into the animals and thus could represent an effective detoxification mechanism. It was shown that a significant lowering of the internal pH occurred within only 20 min after adding sulphide. This pH lowering appeared to be dependent on the external sulphide concentration of the ambient medium and showed a saturation beyond a threshold level of about 130 microM. It is discussed whether this sulphide-induced pH drop is an active regulatory mechanism and acts as an effective protection mechanism against sulphide during short-term exposures.

Animals↗