Ultrastruktur der Liquorkontaktneurone des Zentralkanals des Rückenmarkes vom Karpfen (Cyprinus carpio)

Zusammenfassung Zwischen und unter den Ependymzellen des Zentralkanals des Rückenmarkes vonCyprinus carpio kommen bipolare Nervenzellen vor. Der eine Fortsatz der Neurone (Liquorkontaktneurone) verläuft zwischen den Ependymzellen hindurch, tritt in den Canalis centralis ein und bildet im Liquor cerebrospinalis eine charakteristische Nervenendigung (Liquorkontakt-Nervenendigung). Diese besteht aus einem zentralen Körper und davon radiär ausstrahlenden zahlreichen Stereozilien, sowie einer Kinozilie. Ein direkter Kontakt zwischen den Liquorkontakt-Nervenendigungen und dem Reissnerschen Faden wurde nicht beobachtet. Der ependymofugale Fortsatz der Neurone hat Axonnatur. Im Perikaryon der Nervenzellen findet man endoplasmatisches Retikulum mit glatter und rauher Oberfläche, freie Ribosomen, Golgi-Areale, sowie Mitochondrien und granulierte Vesikel (Durchmesser 700–1000 Å). Axone, die synaptische und granulierte Bläschen (Durchmesser 700–1000 Å) enthalten, bilden mit den Perikaryen und den zum Zentralkanal verlaufenden Nervenfortsätzen Synapsen. Auf den Ependymzellen konnten stellenweise atypische Zilien beobachtet werden. Der Vergleich der bisher untersuchten Tierarten zeigt, daß die Struktur der spinalen Liquorkontaktneurone von den Fischen bis zu den Säugern prinzipiell gleichartig ist.

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Ultrastructure of the CSF contacting neurons of the central canal of the spinal cord in the carp (Cyprinus carpio)

Summary Bipolar nerve cells are situated between and below the ependymal cells of the central canal of the spinal cord ofCyprinus carpio. One of the processes of the neurons (CSF contacting neurons) passes by between the ependymal cells into the central canal and forms there a characteristic nerve ending in the cerebrospinal fluid (CSF contacting nerve ending). These terminals consist of a central body with numerous stereocilia and one kinocilium. We could not observe any direct contact between the CSF contacting nerve endings and Reissner's fibre. The ependymofugal process of the nerve cells is axon-like. In the perikarya of the CSF contacting neurons, dense-core vesicles (diameter 700–1000 Å) are found besides of smooth and rough-surfaced endoplasmic reticulum, free ribosomes, Golgi areas and mitochondria. Axons containing synaptic and granulated vesicles (diameter 700–1000 Å), form synapses with the perikarya and the CSF contacting nerve processes. On the surface of single ependymal cells, atypical cilia could be observed. The comparison of the species studied up to now, shows that the structure of the spinal CSF contacting neurons is principally similar from fishes up to mammals.

     

Lens fibre cell differentiation and organelle loss: many paths lead to clarity

The programmed removal of organelles from differentiating lens fibre cells contributes towards lens transparency through formation of an organelle-free zone (OFZ). Disruptions in OFZ formation are accompanied by the persistence of organelles in lens fibre cells and can contribute towards cataract. A great deal of work has gone into elucidating the nature of the mechanisms and signalling pathways involved. It is apparent that multiple, parallel and redundant pathways are involved in this process and that these pathways form interacting networks. Furthermore, it is possible that the pathways can functionally compensate for each other, for example in mouse knockout studies. This makes sense given the importance of lens clarity in an evolutionary context. Apoptosis signalling and proteolytic pathways have been implicated in both lens fibre cell differentiation and organelle loss, including the Bcl-2 and inhibitor of apoptosis families, tumour necrosis factors, p53 and its regulators (such as Mdm2) and proteolytic enzymes, including caspases, cathepsins, calpains and the ubiquitin-proteasome pathway. Ongoing approaches being used to dissect the molecular pathways involved, such as transgenics, lens-specific gene deletion and zebrafish mutants, are discussed here. Finally, some of the remaining unresolved issues and potential areas for future studies are highlighted.     

Effect of cellobiose supplementation and dietary soluble fibre content on in vitro caecal fermentation of carbohydrate-rich substrates in rabbits

The in vitro caecal fermentation of five substrates low in starch and protein content [d-(+)-glucose (GLU), d-cellobiose (CEL), sugar beet pectin (PEC), sugar beet pulp (SBP) and wheat straw (WS)] was investigated using soft faeces from rabbits receiving different levels of cellobiose and soluble fibre as inoculum. A total of 24 rabbits were supplemented 3 levels of cellobiose in the drinking water (0.0, 7.5, 15.0 g/l) and fed two experimental diets containing either low soluble fibre (LSF) or high soluble fibre (HSF) levels (84.0 and 130 g/kg dry matter). All substrates were subjected to a two-step pepsin/pancreatin in vitro pre-digestion, and the whole residue was used as substrate for the in vitro incubations. Gas production was measured until 144 h, and volatile fatty acid (VFA) production was determined at 24 h incubation. Experimental treatments did not affect SBP fermentation and had only a subtle influence on fermentation of WS and GLU. In contrast, cellobiose supplementation × donors’ diet interactions were detected for most gas production parameters for CEL. Both the fractional gas production (k) and maximal gas production rates were linearly increased (p ≤ 0.042) and the initial delay in the onset of gas production (Lag) linearly decreased (p < 0.001) by cellobiose supplementation with the HSF inoculum, with no differences between the 7.5 and 15.0 doses. In contrast, with the LSF inoculum cellobiose supplementation only affected k values, which were quadratically increased (p = 0.043) and had maximal values for the 7.5 dose. A quadratic effect (p ≤ 0.018) of cellobiose supplementation was observed for total VFA production at 24 h when CEL and PEC were fermented, obtaining the maximal VFA production for the 7.5 dose of cellobiose. Total VFA production for CEL was greater with LSF than with HSF inoculum (20.7 vs. 12.9 mmol/l; p = 0.014), but the opposite was found for WS (3.97 vs. 6.21 mmol/l; p = 0.005). The use of LSF inoculum for CEL fermentation sharply reduced acetate (p = 0.001) and increased butyrate proportions (p ≤ 0.001) compared with the HSF inoculum. A positive relationship between total VFA caecal concentrations in rabbits receiving the same experimental treatments and in vitro values was only observed when WS was used as substrate (r = 0.90; p = 0.015; n = 6). The results suggest that experimental factors influenced the fermentative activity of caecal digesta, but the observed response differed with the incubated substrate, being the CEL the most affected.     

A phylogenetic and functional perspective on the origin and evolutionary shifts of growth ring anatomical markers in seed plants

We reconstruct the evolutionary changes in different anatomical markers in order to understand the evolution and functional aspects of growth rings during the diversification of seed plants (spermatophytes), one of the largest and most diverse lineages of the tree of life. We carried out a wide revision of the anatomy of secondary xylem in spermatophytes and reconstructed the evolution of the different anatomical markers in a time-calibrated phylogeny. By embodying a functionally and evolutionarily significant concept in growth rings we reveal a new panorama for their frequency and show how common they are in diverse lineages of tropical plants. In this context, the principal anatomical markers of growth rings are identified in the evolutionary history of plants and their association with climate-related ecological characteristics. We discuss the function of these anatomical markers, especially for thick-walled and/or radially flattened latewood fibres, fibre zone and dilated rays. Despite the high evolutionary lability of the anatomical markers evidenced by our analyses, they appear to represent deep homologies.     

Differences in kainate receptor involvement in hippocampal mossy fibre long-term potentiation depending on slice orientation

Long-term potentiation (LTP) is a well-established experimental model used to investigate the synaptic basis of learning and memory. LTP at mossy fibre - CA3 synapses in the hippocampus is unusual because it is normally N-methyl-d-aspartate (NMDA) receptor-independent. Instead it seems that the trigger for mossy fibre LTP involves kainate receptors (KARs). Although it is generally accepted that pre-synaptic KARs play an essential role in frequency facilitation and LTP, their subunit composition remains a matter of significant controversy. We have reported previously that both frequency facilitation and LTP can be blocked by selective antagonism of GluK1 (formerly GluR5/Glu(K5))-containing KARs, but other groups have failed to reproduce this effect. Moreover, data from receptor knockout and mRNA expression studies argue against a major role of GluK1, supporting a more central role for GluK2 (formerly GluR6/Glu(K6)). A potential reason underlying the controversy in the pharmacological experiments may reside in differences in the preparations used. Here we show differences in pharmacological sensitivity of synaptic plasticity at mossy fibre - CA3 synapses depend critically on slice orientation. In transverse slices, LTP of fEPSPs was invariably resistant to GluK1-selective antagonists whereas in parasagittal slices LTP was consistently blocked by GluK1-selective antagonists. In addition, there were pronounced differences in the magnitude of frequency facilitation and the sensitivity to the mGlu2/3 receptor agonist DCG-IV. Using anterograde labelling of granule cells we show that slices of both orientations possess intact mossy fibres and both large and small presynaptic boutons. Transverse slices have denser fibre tracts but a smaller proportion of giant mossy fibre boutons. These results further demonstrate a considerable heterogeneity in the functional properties of the mossy fibre projection.     

Isometric and Eccentric Force Generation Assessment of Skeletal Muscles Isolated from Murine Models of Muscular Dystrophies

Critical to the evaluation of potential therapeutics for muscular disease are sensitive and reproducible physiological assessments of muscle function. Because many pre-clinical trials rely on mouse models for these diseases, isolated muscle function has become one of the standards for Go/NoGo decisions in moving drug candidates forward into patients. We will demonstrate the preparation of the extensor digitorum longus (EDL) and diaphragm muscles for functional testing, which are the predominant muscles utilized for these studies. The EDL muscle geometry is ideal for isolated muscle preparations, with two easily accessible tendons, and a small size that can be supported by superfusion in a bath. The diaphragm exhibits profound progressive pathology in dystrophic animals, and can serve as a platform for evaluating many potential therapies countering fibrosis, and promoting myofiber stability. Protocols for routine testing, including isometric and eccentric contractions, will be shown. Isometric force provides assessment of strength, and eccentric contractions help to evaluate sarcolemma stability, which is disrupted in many types of muscular dystrophies. Comparisons of the expected results between muscles from wildtype and dystrophic muscles will also be provided. These measures can complement morphological and biochemical measurements of tissue homeostasis, as well as whole animal assessments of muscle function.     

Effect of disuse on the ultrastructure of the achilles tendon in rats

We examined the influence exerted, through disuse of the hindlimb, on the collagen fibres of the achilles tendon in rats. With disuse the body mass decreased by 28%, and the mass of soleus muscle decreased by 20%. A decrease in the surface area and diameter was observed in the experimental group when compared to the control group. A histogram of the collagen fibres showed a decrease of the thick fibres in the experimental group. The maximum surface area of collagen fibres in the experimental group was seen to be only 43% of that of the control group. These results showed a decrease in the thickness of the collagen fibres of the achilles tendon through disuse. This seemed to suggest that resistance to tension is decreased by disuse.