Elektronenmikroskopische Untersuchungen über Glaskörperrindenzellen und Zonulafasern

Zusammenfassung Es werden folgende Befunde zur Feinstruktur des Glaskörpers beim 16 Tage alten Rattenembryo und der Glaskörperrinde beiderseits der Ora serrata beim 3 Jahre alten, an Retinoblastom erkrankten Kind mitgeteilt:Der Glaskörper des Rattenembryos und die Glaskörperrinde des kindlichen Auges enthalten Fibroblasten. Sie unterscheiden sich nicht von den im Bindegewebe vorkommenden Fibroblasten. Im embryonalen Rattenglaskörper wurden außerdem faserbildende Zellen mit wabiger Struktur des Zytoplasmas beobachtet.Die Fibroblasten der Glaskörperrinde bilden die Fibrillen des Glaskörpergerüstes und der Zonulafasern. Diese Fibrillen zeigen eine deutliche Querstreifung. Die Streifung ist unregelmäßig oder periodisch. Die Länge der Perioden beträgt in unseren Schnitten bei den Glaskörperfibrillen des Rattenembryos und des kindlichen Auges meist etwa 120, seltener 210 A. An den Fibrillenbündeln einer Zonulafaser des kindlichen Auges wurden Perioden von 90–120, 400, 440 und 630 A beobachtet.Die Fibroblasten der Glaskörperrinde des kindlichen Auges liegen im Bereich der Pars plana corporis ciliaris auch tief in den Buchten und Falten des Ciliarepithels. Hierdurch wird eine maximal große Anheftungsfläche für die von ihnen produzierten Fibrillen der Zonulafasern gewährleistet.Die Pars plana corporis ciliaris des kindlichen Auges ist von einem dichten Netz von Fibroblastenfortsätzen überzogen. Auch vereinzelte Makrophagen finden sich hier.Unsere elektronenmikroskopischen Befunde bestätigen die Angaben von Balazs über das Vorkommen von Fibrocyten (Fibroblasten) und Makrophagen in der Glaskörperrinde. Ferner bestätigen sie die bereits von früheren Autoren lichtmikroskopisch gewonnenen Erkenntnisse, wonach es sich beim Glaskörper um mesenchymales Gewebe, bei den Zonulafasern im Bindegewebsfasern handelt.

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Summary The following electron microscopical findings in the vitreous body of 16-day-old rat embryos and in the vitreoretinal border layer on both sides of the ora serrata in a 3-year-old child are reported:The vitreous body of the rat embryo and the vitreoretinal border layer of the infant eye contain fibroblasts. These fibroblasts do not differ from those present in connective tissue. The embryonic vitreous body of the rat contains fibre-forming cells, which show an alveolar structure of the cytoplasm.The fibroblasts in the cortical tissue layer of the vitreous body form the fibrils of the stroma of the vitreous body and the zonula fibres. These fibrils show a marked cross striation. The striation is either irregular or shows periodicity. In the vitreous body of the rat embryo and of the infant eye the lenght of these periods has been measured with 120 and 210 A. Periods of 90–120, 400, 440 and 630 A could be shown in the fibrillar bundles of a zonula fibre of the infant eye. In the region of the pars plana corporis ciliaris the fibroblasts of the cortical tissue layer of the vitreous body of the infant eye are also found deep down in the sinus and folds of the ciliary epithelium. Thus they guarantee an as large as possible area for the attachment of the fibrils of the zonula fibres which they produce.The pars plana corporis ciliaris of the infant eye is covered with a dense network of fibroblast processes. Also cells of the macrophage type may be found in this region.Our electron microscopical findings confirm those of earlier light microscopists and lately by Balazs. The vitreous body of the eye is formed by mesenchymal tissue, whereas the zonula fibres are formed by connective tissue.

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Mit dankenswerter Unterstützung durch den Schweizerischen Nationalfonds zur Förderung der Wissenschaftlichen Forschung.     

Production of doubled haploid lines in frequencies sufficient for barley breeding programs

Winter barley (Hordeum vulgare L.) anthers were cultured on different liquid and on starch-solidified media. The optimal embryo and callus formation with different F₁-lines and the cv. Igri was obtained on a liquid medium with 20% Ficoll, 20 g/l maltose and barley starch. But the influence of the growth conditions of the donor plants and the genotypical differences are still enormous. The procedure has been optimized to such an extent that it can be used economically.     

CGRP-immunoreactive sensory nerve fibers in the submandibular gland of the rat

Summary Indirect immunofluorescence technique was used to study the occurrence and distribution of CGRP immunoreactivity in the submandibular gland of normal rats and after unilateral sensory and sympathetic denervations. In normal rats, CGRP-immunoreactive nerve fibers and nerve trunks were seen around or in close contact with interlobular salivary ducts as well as around small blood vessels of the gland. Occasionally, CGRP-immunoreactive nerve fibers were also detected between or around the acini of the gland.The submandibular ganglia contained CGRP-immunoreactive nerve fibers, but the ganglion cells were not immunoreactive for CGRP. The trigeminal ganglion contained a population of CGRP-immunoreactive, mainly small sized ganglion cells and nerve fibers distributed throughout the ganglion. Unilateral electrocoagulation of the trigeminal nerve caused a significant reduction in the number of immunoreactive nerve fibers in the gland, although some fibers still were present in the ipsilateral glandular tissue. Unilateral superior cervical ganglionectomy caused no detectable effect on the number of CGRP-immunoreactive nerve fibers in the gland.The present results suggest that the rat submandibular gland contains CGRP-immunoreactive nerve fibers both around blood vessels and in glandular secretory elements. Denervation experiments support the view that the majority, but perhaps not all of them originate from the trigeminal ganglion.     

Growth-associated modifications of low-molecular-weight thiols and protein sulfhydryls in human bronchial fibroblasts

The thiol redox status of cultured human bronchial fibroblasts has been characterized at various growth conditions using thiol-reactive monobromobimane, with or without the combination of dithiotreitol, a strong reducing agent. This procedure has enabled measurement of the cellular content of reduced glutathione (GSH), total glutathione equivalents, cysteine, total cysteine equivalents, protein sulfhydryls, protein disulfides, and mixed disulfides. Passage of cells with trypsin perturbs the cellular thiol homeostasis and causes a 50% decrease in the GSH content, whereas the total cysteine content is subsequently increased severalfold during cell attachment. During subsequent culture, transient severalfold increased levels of GSH, protein-bound thiols, and protein disulfides are reached, whereas the total cysteine content gradually declines. These changes in the redox balance of both low-molecular-weight thiols and protein-bound thiols correlate with cell proliferation and mostly precede the major growth phase. When the onset of proliferation is inhibited by maintenance of cells in medium containing decreased amounts of serum, the GSH content remains significantly increased. Subsequent stimulation of growth by addition of serum results in decreased GSH levels at the onset of proliferation. In thiol-depleted medium, proliferation is also inhibited, whereas GSH levels are increased to a lesser extent than in complete medium. Exposure to buthionine sulfoximine inhibits growth, prevents GSH synthesis, and results in accumulation of total cysteine, protein-bound cysteine, and protein disulfides. For extracellular cystine, variable rates of cellular uptake correlate with the initial increase in the total cysteine content observed following subculture and with the GSH peak that precedes active proliferation. The results strongly suggest that specific fluctuations in the cellular redox balance of both free low-molecular-weight thiols and protein sulfhydryls are involved in growth regulation of normal human fibroblasts.     

Starling circannual systems: are they arrested in long photoperiods?

Summary In the European starling,Sturnus vulgaris, circannual rhythms in gonadal size, molt and other related functions persist only in photoperiods close to 12 h, but are absent in longer or shorter daylengths. To find out whether the arrhythmia seen in long photoperiods results from an arrest of the underlying clock system, three groups of male starlings were held for 10, 14, or 20 months in a 13 h photoperiod and then transferred to a 12 h photoperiod. A control group was held in the 13 h photoperiod throughout the experiment for 28 months. During the initial exposure to the 13 h photoperiod, all birds went through a gonadal cycle, followed by a complete molt. Subsequently, the control birds retained small testes to the end of the experiment and there was no further molt. In contrast, most of the experimental birds re-initiated a testicular cycle, following transfer to the 12 h photoperiod and molted after its completion. The latency between the transfer to the 12 h photoperiod and the onset of testicular growth was not significantly different among the three groups, indicating that the underlying circannual clock had been arrested in the 13 h photoperiod and restarted in the 12 h photoperiod. The pattern of the second testicular cycle did, however, differ among groups. Particularly its amplitude decreased from group 1 to group 3, suggesting that the capacity of the birds to respond to a 12 h photoperiod decreased with increasing duration of exposure to the 13 h photoperiod.Dedicated to Prof. Dr. C.S. Pittendrigh on the occasion of his seventieth birthday.     

Age-dependent modifications of mitochondrial proteins in cerebral cortex and striatum of rat brain

The protein composition of free mitochondria purified from cerebral cortex and striatum during aging was analyzed by gel electrophoresis. Mitochondria were isolated from cerebral cortex and striatum of 4-, 12-, and 24-month-old rat brain. The percent amount of mitochondrial proteins after gel-electrophoretic separation was determined densitometrically. A significant decrease in the amount of two polypeptides (with molecular weights of 20 and 16 kDa, respectively) in both brain regions during aging was found. The decrease was higher in the striatum indicating a greater vulnerability of this brain area to the aging process. The age-dependent modifications of mitochondrial proteins observed may play an important role in several mitochondrial functions, such as energy transduction and transport processes as well as in structural changes occurring with age, causing altered membrane permeability and fluidity.     

On the ultrastructure and the supposed function of the mineralizing matrix coat of sea urchins (Echinodermata,Echinoida)

Summary Echinoderm ossicles are part of the mesenchyme. Their formation and growth, with respect to the underlying tissues, is studied using echinoid spines and teeth and applying different methods of fixation. The calcification process in echinoderms is strictly intracellular and needs (1) syncytial sclerocytes which completely enclose (2) a vacuolar cavity which in turn contains (3) an organic matrix coat. Strictly speaking, each ossicle is nothing but the calcified vacuolar space of a single syncytium of sclerocytes. In fully grown parts, however, the continuous sheath may split open and the matrix-coated mineral may come into contact with the extracellular space. According to biochemical analyses the matrix consists of insoluble components, but most (95%) of its constituents are soluble in EDTA or weak acids. If routine transmission electron microscope methods are used the soluble components are lost and the matrix at best looks electron light. If tannic acid is added to the fixative the soluble matrix components are preserved and reveal further ultrastructural details of the biomineralization process in echinoderms. The matrix coat looks extremely electron dense. Further soluble material is to be found within the vacuolar space or attached to the vacuolar surface of the cytoplasmic sheath. The results lead to the opinion that the matrix coat consists of a hydrophobic framework of insoluble components that contains soluble components which guide the Ca through pores in the hydrophobic layers into the interior of the matrix-coated space. It is only within this space that the mineral is deposited.