Architecture of the nervous system in mystacocarida (Arthropoda,crustacea)—An immunohistochemical study and 3D reconstruction

Mystacocarida is a species‐poor group of minute crustaceans with unclear phylogenetic affinities. Previous studies have highlighted the putative “primitiveness” of several mystacocarid features, including the architecture of the nervous system. Recent studies on arthropod neuroarchitecture have provided a wealth of characters valuable for phylogenetic reconstructions. To permit and facilitate comparison with these data, we used immunohistochemical labeling (against acetylated α‐tubulin, serotonin and FMRFamide) on the mystacocarid Derocheilocaris remanei, analyzing it with confocal laser‐scanning microscopy and 3D reconstruction. The mystacocarid brain is fairly elongated, exhibiting a complicated stereotypic arrangement of neurite bundles. However, none of the applied markers provided evidence of structured neuropils such as a central body or olfactory glomeruli. A completely fused subesophageal ganglion is not present, all segmental soma clusters of the respective neuromeres still being delimitable. The distinct mandibular commissure comprises neurite bundles from more anterior regions, leading us to propose that it may have fused with an ancestral posterior tritocerebral commissure. The postcephalic ventral nervous system displays a typical ladder‐like structure with separated ganglia which bears some resemblance to larval stages in other crustaceans. Ganglia and commissures are also present in the first three limbless “abdominal” segments, which casts doubt on the notion of a clear‐cut distinction between thorax and abdomen. An unpaired longitudinal median neurite bundle is present and discussed as a potential tetraconate autapomorphy. Additionally, a paired latero‐longitudinal neurite bundle extends along the trunk. It is connected to the intersegmental nerves and most likely fulfils neurohemal functions. We report the complete absence of serotonin‐ir neurons in the ventral nervous system, which is a unique condition in arthropods and herein interpreted as a derived character. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

The formation of the nervous system during larval development in Triops cancriformis (Bosc) (crustacea,Branchiopoda): An immunohistochemical survey

We provide data of the development of thenervous system during the first five larval stages of Triops cancriformis. We use immunohistochemical labeling (against acetylated α‐tubulin, serotonin, histamine, and FMRFamide), confocal laser scanning microscopy analysis, and 3D‐reconstruction. The development of the nervous system corresponds with the general anamorphic development in T. cancriformis. In larval stage I (L I), all brain parts (proto‐, deuto‐, and tritocerebrum), the circumoral connectives, and the mandibular neuromere are already present. Also, the frontal filaments and the developing nauplius eye are already present. However, until stage L III, the nauplius eye only consists of three cups. Throughout larval development, the protocerebral network differentiates into distinct subdivisions. In the postnaupliar region, additional neuromeres and their commissures emerge in an anteroposterior gradient. The larval nervous system in L V consists of a differentiated protocerebrum including a central body, a nauplius eye comprising four cups, a circumoral nerve ring, mandibular‐ and postnaupliar neuromeres up to the seventh thoracic segment, each featuring an anterior and a posterior commissure, and two parallel connectives. The presence of a protocerebral bridge is questionable. The distribution of neurotransmitters in L I is restricted to the naupliar nervous system. Over the course of the five stages of development, neurotransmitter distribution also follows an anteroposterior gradient. Each neuromere is equipped with two ganglia innervating the locomotional appendages and possesses a specific neurotransmitter distribution pattern. We suggest a correlation between neurotransmitter expression and locomotion. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Adaptation of Bathynellacea (Crustacea,Syncarida) to Life in the Interstitial (“Zoea Theory”)

This study aims at demonstrating that the Bathynellacea arose by neoteny from a zoea-like ancestor (“Zoea Theory”). Bathynellacea basically have a larval morphology. Their post-embryonic development passes through phases which are directly comparable with some in the development of primitive Eucarida. These phases are called the parazoeal and bathynellid phase, and correspond to the protozoeal and zoeal phase of Penaeidea, respectively. Running initially parallel to that of Penaeidea, the development of the Bathynellacea breaks off precociously. They reach sexual maturity at a stage which, in the Penaeidea, is followed by metamorphosis. There are reasons to assume that the development of all Syncarida originally passed through a series of free-living larval stages and underwent metamorphosis. The bathynellaceans reach adulthood at a stage which corresponds to the last larval stage of their ancestor. The ecological path to interstitial life of the Bathynellacea and the bearing of the “Zoea Theory” upon views as to their phylogenetic position are discussed. The classical example of a group of crustaceans thought to have arisen by neoteny, the Cladocera, is critically examined and found to be in need of reevaluation.     

Novel aspects of the nervous system of Bonellia viridis (Echiura) revealed by the combination of immunohistochemistry, confocal laser-scanning microscopy and three-dimensional reconstruction

The Echiura have been placed in close phylogenetic affinity to the Annelida on the basis of numerous homologous characters including the mode of development, the nearly identical formation of a trochophore larva, as well as the development and ultrastructure of chaetae and spermatozoa. Furthermore, phylogenetic analysis of elongation factor-1 gene sequences supports placement of the Echiura within the Annelida. Nevertheless, the Echiura are generally excluded from the Annelida due to their lack of segmentation. However, it must be considered that this lack could represent a secondary condition and that Echiura are derived from formerly segmented ancestors. In the present study, the combination of methods applied reveals several novel aspects of the central nervous system in developmental stages of Bonellia viridis. The most important of these is the metameric organization of the ventral nerve cord. Antibodies against different neurotransmitters label discrete repetitive units of perikarya in the ventral nerve cord. This organisation is additionally supported by the distribution of peripheral nerves as shown by labelling of neurotubules. These nerves are clearly paired and are evenly distributed, corresponding to the serial units of serotoninergic neurons. Different methods of computer-aided three-dimensional reconstruction display the precise spatial distribution of perikarya and peripheral nerves allowing the repetitive units to be discerned on the basis of relative size, position and number of labelled cells. The repetitive units in the nervous system of B. viridis correspond to segmental ganglia of various Annelida and are interpreted as an indication that Echiura are derived from formerly segmented ancestors, thus supporting the systematic inclusion of the Echiura within the Annelida.     

Uptake of serotonin, 5-hydroxytryptophan and tryptophan by giant serotonin-containing neurones and other neurones in the central nervous system of the snail (Helix pomatia)

Summary Following exposure to tritiated 5-HTP, silver grains were observed over the perikarya of the GSCs (Giant serotonin cells) of Helix pomatia and other known serotonin-containing neurones in light and electron microscope autoradiograms. There was no indication that the 5-HTP was taken up by nerve endings or by non-nervous structures. The distribution of radioactivity was completely different in autoradiograms of tissue exposed to tritiated serotonin. Silver grains, often in very high concentrations, were observed only over certain fine axon branches and processes thought to be nerve endings. Electron microscope autoradiography showed that these processes contained small dense-cored vesicles, morphologically identical to those thought to sequester serotonin in the perikarya of the GSCs. The accumulation of tritiated tryptophan was less specific; all the neurone perikarya showed an accumulation of radioactivity after exposure to this substance.We are grateful to Professor J. F. Lamb for the use of the Scintillation Spectrometer.     

Inhibition of development, swarming differentiation and virulence factors in Proteus mirabilis by an extract of Lithrea molleoides and its active principle (Z,Z)-5-(trideca-4’,7’-dienyl)-resorcinol

Antibacterial activity of Lithrea molleoides extract against Proteus mirabilis has been previously reported by our group. In the present study, the compound (Z,Z)-5-(trideca-4’,7’-dienyl)-resorcinol (1) was isolated as its responsible active principle. The effects of the compound obtained and of L. molleoides extract on P. mirabilis growth and virulence factors were evaluated.Compound 1 showed MIC and MBC values of 4000 μg/ml. It was found that the extract, at four times the MIC, produced complete killing of the uropathogen at 2 h from the beginning of the experiment, while the alkylresorcinol, at four times the MIC, produced the same effect after 24 h. Hemolysis was adversely affected in treatments with both products at 8 μg/ml, while hemagglutination was not altered. The whole extract induced complete autoaggregation of P. mirabilis at 2000 μg/ml, while compound 1 at the same concentration did not show this property. Swarming motility was delayed in treatments with the extract and with 1 at 1000 and 8 μg/ml, respectively, at 8 h from the beginning of the assay. Complete inhibition of the phenomenon was still observed after 24 h when compound 1 was added at 125 μg/ml.These findings offer the possibility of new classes of antimicrobial medicines to tackle infections caused by P. mirabilis.