scholarly journals Nervous system and ciliary structures of Micrognathozoa (Gnathifera): evolutionary insight from an early branch in Spiralia

2016 ◽  
Vol 3 (10) ◽  
pp. 160289 ◽  
Author(s):  
Nicolas Bekkouche ◽  
Katrine Worsaae
Keyword(s):  
Nervous System ◽  
Laser Scanning ◽  
Complex Structure ◽  
Sister Group ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Large Brain ◽  
Close Relationship ◽  
The Many ◽  
Collecting Tubules

Recent studies show that Gnathifera, comprising Rotifera, Gnathostomulida and Micrognathozoa, constitute the sister group to the remaining Spiralia (containing, e.g. flatworms, segmented worms and molluscs). Therefore, a better understanding of Gnathifera is central for unravelling the evolution of the highly diverse Spiralia. Here, we describe the previously unstudied nervous system and ciliary structures of Micrognathozoa, using immunohistochemistry and confocal laser scanning microscopy. The nervous system is simple with a large brain, paired sub-esophageal ganglia, two trunk commissures, two pairs of ventral longitudinal nerves and peripheral nerves. The paired ventro-lateral nerve cords are confirmed to be a symplesiomorphy of Gnathifera (possibly even Spiralia), whereas the paired ventro-median nerves are not previously reported in Gnathifera. A pharyngeal ganglion is described for Micrognathozoa: a complex structure with two apical tufts of ciliary receptors, now shown to be shared by all Gnathifera. The ventral pattern of external ciliophores is re-described, and protonephridia with multi-ciliated collecting tubules similar to those of Rotifera are confirmed. A range of new details from a simple nervous system and complex set of ciliary structures in a microscopic metazoan are hereby unravelled. The many resemblances with Rotifera corroborate their close relationship, and shed more light on the evolution of Gnathifera.

Confocal Laser Microscopy of Physiologically Characterized Fluorescently Labeled Central Nervous System Neurons

1988 ◽  
Vol 46 ◽  
pp. 274-275
Author(s):  
J.N. Turner ◽  
J. Swann ◽  
K. Smith ◽  
M. Siemens ◽  
D. Szarowski ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Laser Scanning ◽  
Tissue Sample ◽  
Lucifer Yellow ◽  
Single Cells ◽  
Brain Slices ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Efficient Detection

Confocal laser scanning microscopy (CLSM) is capable of three-dimensional imaging of fluorescently labeled single cells. Efficient detection via a photomultiplier and optical sectioning with high rejection of light from other specimen levels make it possible to image cells surrounded by either labeled or unlabeled tissue. It is no longer necessary to restrict high resolution light microscopy to cultured cells or those near the surface of a tissue sample. Cells can be observed üin situ” in a physiologically characterized environment. Central nervous system neurons can be electrophysiologically characterized and then injected with a fluorescent dye such as lucifer yellow. The CLSM can excite the dye and image the fluorescent emission in thick tissue preparations (hundreds of micrometers) making possible a new approach to the correlation of physiology and anatomy.Brain slices 350 μm thick were obtained from hippocampus and inferior colliculus of immature rats and incubated in oxygenated artificial cerebrospinal fluid. Cells were penetrated with micropipets, characterized electrophysiologically and ionophoretically injected with 5% lucifer yellow in LiAc.

scholarly journals Heparin Inhibits Leukocyte Rolling in Pial Vessels and Attenuates Inflammatory Changes in a Rat Model of Experimental Bacterial Meningitis

1997 ◽  
Vol 17 (11) ◽  
pp. 1221-1229 ◽  
Author(s):  
Joerg R. Weber ◽  
Klemens Angstwurm ◽  
Thomas Rosenkranz ◽  
Ute Lindauer ◽  
Dorette Freyer ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Bacterial Meningitis ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Leukocyte Rolling ◽  
Central Nervous System Inflammation ◽  
Inflammatory Processes

Heparin is a natural proteoglycan that was first described in 1916. In addition to its well characterized effect on blood coagulation, it is becoming clear that heparin also modulates inflammatory processes on several levels, including the interference with leukocyte–endothelium interaction. Anecdotal observations suggest a better clinical outcome of heparin-treated patients with bacterial meningitis. The authors demonstrate that heparin, a glycosaminoglycan, inhibits significantly in the early phase of experimental pneumococcal meningitis the increase of 1) regional cerebral blood flow (125 ± 18 versus 247 ± 42%), 2) intracranial pressure (4.5 ± 2.0 versus 12.1 ± 2.2 mm Hg), 3) brain edema (brain water content: 78.23 ± 0.33 versus 79.49 ± 0.46%), and 4) influx of leukocytes (571 ± 397 versus 2400 ± 875 cells/μL) to the cerebrospinal fluid compared with untreated rats. To elucidate the possible mechanism of this observation, the authors investigated for the first time leukocyte rolling in an inflammatory model in brain venules by confocal laser scanning microscopy in vivo. Heparin significantly attenuates leukocyte rolling at 2, 3, and 4 hours (2.8 ± 1.3 versus 7.9 ± 3.2/100 μm/min), as well as leukocyte sticking at 4 hours (2.1 ± 0.4 versus 3.5 ± 1.0/100 μm/min) after meningitis induction compared with untreated animals. The authors conclude that heparin can modulate acute central nervous system inflammation and, in particular, leukocyte–endothelium interaction, a key process in the cascade of injury in bacterial meningitis. They propose to evaluate further the potential of heparin in central nervous system inflammation in basic and clinical studies.

scholarly journals Morphology of Stephanella hina (Bryozoa, Phylactolaemata): common phylactolaemate and unexpected, unique characters

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Thomas F. Schwaha ◽  
Masato Hirose
Keyword(s):  
Laser Scanning ◽  
Mouth Opening ◽  
Morphological Characteristics ◽  
Sister Group ◽  
Neurosecretory Cells ◽  
Laser Scanning Microscopy ◽  
Immunocytochemical Staining ◽  
Confocal Laser ◽  
Muscle Fibres ◽  
Microscopy Techniques

Abstract Stephanella hina is a little studied freshwater bryozoan belonging to Phylactolaemata. It is currently the only representative of the family Stephanellidae, which in most reconstructions is early branching, sometimes even sister group to the remaining phylactolaemate families. The morphological and histological details of this species are entirely unknown. Consequently, the main aim of this study was to conduct a detailed morphological analysis of S. hina using histological serial sections, 3D reconstruction, immunocytochemical staining and confocal laser scanning microscopy techniques. The general morphology is reminiscent of other phylactolaemates; however, there are several, probably apomorphic, details characteristic of S. hina. The most evident difference lies in the lophophoral base, where the ganglionic horns/extensions do not follow the traverse of the lophophoral arms but bend medially inwards towards the mouth opening. Likewise, the paired forked canal does not fuse medially in the lophophoral concavity as found in all other phylactolaemates. Additional smaller differences are also found in the neuro-muscular system: the rooting of the tentacle muscle is less complex than in other phylactolaemates, the funiculus lacks longitudinal muscles, the caecum has smooth muscle fibres, latero-abfrontal tentacle nerves are not detected and the medio-frontal nerves mostly emerge directly from the circum-oral nerve ring. In the apertural area, several neurite bundles extend into the vestibular wall and probably innervate neurosecretory cells surrounding the orifice. These morphological characteristics support the distinct placement of this species in a separate family. Whether these characteristics are apomorphic or possibly shared with other phylactolaemates will require the study of the early branching Lophopodidae, which remains one of the least studied taxa to date.

scholarly journals Translational Research for Orthopedic Bone Graft Development

Materials ◽  
2021 ◽  
Vol 14 (15) ◽  
pp. 4130
Author(s):  
Maria J. C. Vilela ◽  
Bruno J. A. Colaço ◽  
José Ventura ◽  
Fernando J. M. Monteiro ◽  
Christiane L. Salgado
Keyword(s):  
Production Process ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Laser Scanning ◽  
Complex Structure ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Mechanical Resistance ◽  
Human Mscs ◽  
Biological Studies

Designing biomaterials for bone-substitute applications is still a challenge regarding the natural complex structure of hard tissues. Aiming at bone regeneration applications, scaffolds based on natural collagen and synthetic nanohydroxyapatite were developed, and they showed adequate mechanical and biological properties. The objective of this work was to perform and evaluate a scaled-up production process of this porous biocomposite scaffold, which promotes bone regeneration and works as a barrier for both fibrosis and the proliferation of scar tissue. The material was produced using a prototype bioreactor at an industrial scale, instead of laboratory production at the bench, in order to produce an appropriate medical device for the orthopedic market. Prototypes were produced in porous membranes that were e-beam irradiated (the sterilization process) and then analysed by scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), dynamic mechanical analysis (DMA), cytotoxicity tests with mice fibroblasts (L929), human osteoblast-like cells (MG63) and human MSC osteogenic differentiation (HBMSC) with alkaline phosphatase (ALP) activity and qPCR for osteogenic gene expression. The prototypes were also implanted into critical-size bone defects (rabbits’ tibia) for 5 and 15 weeks, and after that were analysed by microCT and histology. The tests performed for the physical characterization of the materials showed the ability of the scaffolds to absorb and retain water-based solvents, as well as adequate mechanical resistance and viscoelastic properties. The cryogels had a heteroporous morphology with microporosity and macroporosity, which are essential conditions for the interaction between the cells and materials, and which consequently promote bone regeneration. Regarding the biological studies, all of the studied cryogels were non-cytotoxic by direct or indirect contact with cells. In fact, the scaffolds promoted the proliferation of the human MSCs, as well as the expression of the osteoblastic phenotype (osteogenic differentiation). The in vivo results showed bone tissue ingrowth and the materials’ degradation, filling the critical bone defect after 15 weeks. Before and after irradiation, the studied scaffolds showed similar properties when compared to the results published in the literature. In conclusion, the material production process upscaling was optimized and the obtained prototypes showed reproducible properties relative to the bench development, and should be able to be commercialized. Therefore, it was a successful effort to harness knowledge from the basic sciences to produce a new biomedical device and enhance human health and wellbeing.

Streptoneury Is Independent From Ontogenetic Torsion in the Caenogastropod Snail Marisa Cornuarietis

2021 ◽  
Author(s):  
J. Anton Morath ◽  
Stefan Fischer ◽  
Leonie Hannig ◽  
Simon Schwarz ◽  
Rita Triebskorn ◽  
...  
Keyword(s):  
Developmental Biology ◽  
Nervous System ◽  
3D Reconstruction ◽  
Laser Scanning ◽  
Mantle Cavity ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Apple Snail ◽  
Serial Sectioning ◽  
Marisa Cornuarietis

Abstract A hallmark in snails’ anatomy is the conspicuous crossing of the pleurovisceral nerve cords present in all but the most derived gastropod clades. This feature is called streptoneury and hitherto near-universally believed to derive from the process of torsion which is, ontogenetically, visible by a 180° rotation of the visceral sac relative to the cephalopodium, being also responsible for the formation of a cranially bent gut and the location of gills in a mantle cavity that opens to the anterior. However, the mechanical link between the ontogenetic rotation of the visceropallium and streptoneury has never been demonstrated directly. After suppressing ontogenetic torsion in the freshwater apple snail Marisa cornuarietis, we could show in a 3D reconstruction based on serial sectioning that the nervous system of the non-torted snail almost identically mirrored the classical organization of a normally developed individual and showed all features of streptoneury in this species. Furthermore, confocal laser scanning microscopy revealed the pleurovisceral cords not to be fully shaped after completion of ontogenetic torsion. We therefore conclude that, ontogenetically, and potentially also phylogenetically, torsion is not an implicit prerequisite for streptoneury, thereby fundamentally challenging a century-old ‘certainty’ in molluscan developmental biology and evolution.

Phylogeny, evolution and systematic revision of the mite harvestman family Neogoveidae (Opiliones Cyphophthalmi)

2019 ◽  
Author(s):  
Ligia R. Benavides ◽  
Gustavo Hormiga ◽  
Gonzalo Giribet
Keyword(s):  
Laser Scanning ◽  
Phylogenetic Analyses ◽  
Sister Group ◽  
Molecular Study ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Sister Group Relationship ◽  
Systematic Revision ◽  
The Family ◽  
The Usa

Mite harvestmen of the family Neogoveidae have a tropical trans-Atlantic distribution with representatives in equatorial West Africa and the Neotropics, specifically in the south-east region of the USA and in northern South America, being conspicuously absent from Central America. We provide a comprehensive molecular phylogeny of the family including representatives of all genera but the monotypic Tucanogovea Karaman, 2013, and new information on the type species described by Jochen Martens in 1969 that were unavailable for molecular study until now: Brasiliogovea microphaga, Metagovea oviformis and ‘? Gen. enigmaticus’. Additionally, we revisit the somatic and male genitalic morphology of representatives of all genera by means of scanning electron microscopy and confocal laser scanning microscopy, and describe the new genera Leggogovia Benavides & Giribet, gen. nov., Microgovia Benavides, Hormiga & Giribet, gen. nov., Waiwaigovia Benavides, Hormiga & Giribet, gen. nov. and 13 new species: Brasiliogovea aphantostylus Benavides, Hormiga & Giribet, sp. nov., Brasiliogovea microstylus Benavides, Hormiga & Giribet, sp. nov., Brasiliogovea yacambuensis Benavides, Hormiga & Giribet, sp. nov., Metagovea matapi Benavides, Hormiga & Giribet, sp. nov., Metagovea planada Benavides, Hormiga & Giribet, sp. nov., Microgovia chenepau Benavides, Hormiga & Giribet, sp. nov., Neogovea branstetteri Benavides, Hormiga & Giribet, sp. nov., Neogovea enigmatica Martens, sp. nov., Neogovea matawai Benavides, Hormiga & Giribet, sp. nov., Parogovia montealensis Benavides & Giribet, sp. nov., Parogovia prietoi Benavides & Giribet, sp. nov., Parogovia putnami Benavides & Giribet, sp. nov. and Waiwaigovia schultzi Benavides, Hormiga & Giribet, sp. nov. Phylogenetic analyses based on maximum likelihood, parsimony and Bayesian inference support the monophyly of Neogoveidae and a sister group relationship of Neogoveidae + Ogoveidae with Troglosironidae (a clade named Sternophthalmi). Relationships among neogoveid genera are largely congruent between methods as follows: ((Leggogovia gen. nov., Metasiro), (Parogovia, ((Canga, Microgovia gen. nov.), ((Brasiliogovea, Neogovea), (Huitaca, (Waiwaigovia gen. nov., Metagovea)))))). In light of our results, the following taxonomic changes are proposed: Metagovea oviformis Martens, 1969 is transferred to Microgovia, gen. nov.; Parogovia pabsgarnoni Legg, 1990 is transferred to Leggogovia, gen. nov.; ‘? Gen. enigmaticus Martens, 1969’ is an invalid name according to the ICZN; the corresponding taxon is redescribed and formally named as Neogovea enigmatica Martens, sp. nov.

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