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The karyotype of D. elongatus was investigated by means of C-banding, silver staining, and mithramycinand quinacrine fluorescent staining. The diploid chromosome number is 2n = 50. C-banding shows pericentromerically localized constitutive heterochromatin in every chromosome. Two of the chromosome pairs carry two telomeric nucleolus organizer regions each. No heteromorphic sex chromosomes were found.
Behavioural adaptations have made the desert isopod Hemilepistus reaumuri the most successful herbivore and detritivore of the macrofauna of many arid areas in North Africa and Asia Minor. For survival and reproduction Hemilepistus is dependent on burrows. New burrows can only be dug during spring. With the time-consuming digging of a burrow, Hemilepistus has only made the first step towards solving its ecological problems. The burrows are vital and have to be continuously defended against competitors. This requirement is met by co-operation of individuals within the framework of a highly developed social behaviour. In spring adults form monogamous pairs in which partners recognize each other individually and later form, with their progeny, strictly closed family communities. Hemilepistus is compared with a Porcellio' sp. which has developed, convergently, a social behaviour which resembles that of Hemilepistus in many respects, but differs essentially in some aspects, partly reflecting differences in ecological requirements. This and a few other Porcellio species demonstrate some possible steps in the evolution of the social behaviour of Hemilepistus. The female Hemilepistus is-in contrast to Porcellio sp. - semelparous and the selective advantages of monogamy in its environment are not difficult to recognize. This chapter discusses how this mating system could have evolved and especially why monogamous behaviour is also the best method for the Hemilepistus male to maximize its reproductive success. The cohesion of pairs and of family communities in Hemilepistus is based on a highly developed chemical communication system. Individual- and family-specific badges owe their specificity to genetically determined discriminating substances. The nature of the badges raises a series of questions: e.g. since alien badges release aggression, how do parents avoid cannibalizing their young? Similar problems arise from the fact that family badges are mixtures of chemical compounds of very low volatility with the consequence that they can only be transferred by direct contact and that during moulting all substances are lost which an individual does not produce itself. It is shown that in solving these problems inhibiting properties (presumably substances) and learning play a dominant role.
Individual recogmtlon in the non-eusocial arthropods is, according to our present knowledge, predominantly found in the frame of permanent or temporary monogamy. In some cases, e. g. in stomatopods and possibly other marine crustaceans too, individual recognition may serve to allow identification of (i) individuals within dominance hierarchies or (ii) neighbours in territorial species thus helping to avoid the repetition of unnecessary and costly fights. Kin recognition is experimentally proven only in some isopod species (genera Hemilepistus and Porcel/io) and in the primitive cockroach (termite?) Cryptocercus. The «signatures» or «discriminators» used in the arthropods are chemical. It is assumed that the identifying substances are mainly genetically determined and in this paper I shall discuss possible evolutionary origins. The main part of this account is devoted to the presentation of some aspects of the highly developed individual and kin identification and recognition system in the desert isopod Hemilepistus reaumuri - a pure monogamous species in which pairs together with their progeny form strictly exclusive family units. Amongst other things problems of (i) mate choice, (ii) learning to recognize a partner, (iii) avoiding the un adaptive familiarization with aliens are treated. Monogamy under present conditions is for both sexes the only suitable way of maximizing reproductive success; an extremely strong selection pressure must act against every attempt to abandon monogamy under the given ecological conditions. The family «badges» which are certainly always blends of different discriminator substances are extremely variable. This variability is mainly due to genetical differences and is not environmentally caused. It is to be expected that intra-family variabiliry exists in respect of the production of discriminator substances. Since the common badge of a family is the result of exchanging and mixing individual substances, and since the chemical nature of these discriminators requires direct body contacts in order to acquire those substances which an individual does not produce itself, problems must arise with molting. These difficulties do indeed exist and they are aggravated by the fact that individuals may produce substances which do not show up in the common family badge. An efficient learning capability on the one hand and the use of inhibiting properties of newly molted isopods help to solve these problems. In the final discussion three questions are posed and - partly at least - answered; (i) why are families so strictly exclusive, (ii) how many discriminator substances have to be produced to provide a variability allowing families to remain exclusive under extreme conditions of very high population densities, (iii) what is the structure of the family badge and what does an individual have to learn apart from the badge in order not to mistake a family member for an alien or vice versa.
No abstract available
Observations on captive reed frogs Hyperolius viridijlavus ommatostictus showed that seven out of 24 females changed into males. Sex change occurred without any hormone treatment and resulted in completely functional males. The adaptive value is discussed in terms of maximizing life-time reproductive success. Hyperolius r. ommatostictus is the first amphibian known to show functional sex reversal.
The relationship between different degrees of intraspecific crowding of reedfrog tadpoles and their physiological responses to a deterioration of the natal pond water quality was examined under laboratory conditions. Tadpoles that were reared at a lower density metamorphosed significantly earlier than those raised at a higher density. As density increases, the average body length at metamorphosis decreases. However, at low tadpole density, a significantly higher diversity of body size classes among freshly metamorphosed froglets was observed than under more crowded conditions. Mortality increased during metamorphic climax and was inversely correlated with the tadpole density. In ephemeral ponds, an accumulation of nitrogenous wastes from metabolic processes and/or a concentration by evaporation in prolonged rainless periods can pose a considerable chemical stress to reedfrog tadpoles. Hyperolius viridiflavus ommatostictus responded to an increasing ammonia concentration with an activity increase of the ornithine cycle (intensified urea synthesis). hi contrast, Hyperolius marmoratus taeniatus exhibited a strong tolerance against high ammonia levels. A deterioration of the natal pond water quality caused H. v. ommatostictus and H. v. nitidulus tadpoles to adjust to harsher climatic conditions at the time of metamorphosis. This physiological preadjustment enabled the froglets to start feeding and growing immediately after metamorphosis even at low air humidity and rare precipitation events. In contrast, froglets that were raised in daily refreshed water exhibited high mortality rates if subjected to identical conditions. As one possible indicator of the actual climatic conditions prevailing in the surrounding terrestrial habitat, fluctuations in the water ammonia level are discussed.
We have studied the lowest magnetic excitation of Ni\(_{1-x}\)Co\(_x\)O mixed crystals for 0.94 \(\leq\) x \(\leq\) 1. Together with previous results for 0.02 \(\leq\) x \(\leq\) 0.07 and neutron data for x = 0.14 and x = 0.30, the results are discussed by means of a model, especially the variation of AFMR frequency and preferred spin direction with Co concentration x.
The preparation of Bi2Sea single crystals with low free carrier densities allowed an investigation of the lattice vibrations to be carried out from the reflectivity of cleavage planes at nearly normal incidence of the radiation (E ~ c). The experimental results ean be explained with two classical oscillators, whose eigenfrequencies oceur at 92 and 69.5 cm-I. The static dielectric constant was determined to be 100 ± 10 for E ~ c.
The photoconductivity of both undeformed and deformed Te samples has been investigated at liquid He temperature by means of a Fourier spectrometer. Three peaks were usually found in the spectra of undeformed samples at 11, 24 and 46 cm\(^{-1}\). These are shown to be due to three different chemical impurities. The deformed samples are characterized by additional structure at higher frequencies.
The observed impurity induced far-infrared absorption in CsCl : Rb\(^+\) and CsCl : K\(^+\) is compared with a calculated density of acoustic phonon states in CsCl. The absorption due to CsCl : Rb\(^+\) displays a minimum between the acoustic and optic phonon bands. A narrow line is observed in CsCl: K\(^+\) at 85.8 cm\(^{-1}\) which falls in this quasi-phonon gap.
Precipitating anti-PM-Sel antibodies are present in sera from patients with polymyositis. scleroderma. and polymyositis/scleroderma overlap syndromes. By indirect immunofluorescence microscopy. anti-PM-Scl antibodies stained the nucleolus in cells of different tissues and species. suggesting that the antigen is highly conserved. By electron microscopy, anti-PM-Scl antibodies reacted primarily with the granular component of the nuc1eolus. Drugs that inhibit rRNA synthesis had a marked effect on the expression of PM-Scl antigen. In actinomycin D-treated cells, immunofluorescence staining by anti-PM-Scl was signüicantly reduced with residual staining restricted to the granular regions of nuc1eoli. Treatment with 5,6-dichloro-beta-D- ribofuranosylbenzimidazole (DRB) also selectively reduced nuc1eolar staining. On a molecular level, anti-PM-Sel antibodies precipitated 11 polypeptides with molecular weights (Mr) ranging from 110,000 to 20,000. The Mr 80,000 and 20.000 polypeptides were phosphorylated. Evidence suggests that the PM-Scl antigen complex may be related to a prerlbosomal particle.
Antibodies to calf thymus histone H2B were purified by chromatography on DEAE-cellulose and injected into oocyte nuclei of Pleurodeles waltlii. As shown by indirect immunofluorescence these antibodies cross-reacted strongly with corresponding histones associated with lampbrush chromosomes. Shortly after injection the lateral loops of the chromosomes retracted into the chromomeres and by 3 h postinjection the 'lampbrush' appearance was completely lost and the chromosomes appeared in light-microscopic preparations as rod-like structures consisting of 10ngitudina11y coalesced chromomeres. In control oocytes injected with non-immune immunoglobulins or antibodies against a ubiquitous transcript-associated protein no morphological alterations of the lampbrush chromosomes could be observed. Electron microscopic spreads of chromosomes prepared at various times after injection of anti-H2B revealed a progressive loss of transcriptional complexes from the loop axes. Finally, higher-order chromatin configurations, like supranuc1eosomal globules (' superbeads ') or cable-like chromatin strands 50- 60 nm thick predominated, indicating complete transcriptional inactivation of a11 chromosomal regions. The results indicate that H2B antibodies react specifically with his tones associated with the transcribed DNA of lateral loops in their native state. The resulting antigenantibody complexes seem to inhibit progression of the R A polymerases along the template, thus causing the premature release of transcripts, a process analogous to the stripping effect of actinomycin D. The demonstration of histones associated with heavily transcribed regions, which are not compacted into nucleosomes but largely extended, supports the current concept that unfolding of nucleosomes to a110w transcription of the DNA does not involve dissociation of histones. In contrast, amplified ribosomal RNA genes are unaffected by injected HzB antibodies. This does not necessarily indicate absence of his tones from nucleolar chromatin, since we do not know whether it is accessible in vivo to antibodies or whether the histone antigenie determinants are masked by the presence of other proteins. The technique of injecting specific antibodies should be widely applicable when analysing the in vivo distribution of chromosomal components at the electron-microscopic level and when studying complex metabolie processes, like the cleavage and modification of RNA, by selective inhibition of defined enzymic steps.
Autoantibodies to components of the nucleolus are a unique serological feature of patients with scleroderma. There are autoantibodies of several specificities; one type produces a speckled pattern of nucleolar staining in immunofluorescence. In actinomycin D and 5,6-dichloro-{j-D-ribofuranosylbenzimidazoletreated Vero cells, staining was restricted to the fibrillar and not the granular regions. By double immunofluorescence, specific rabbit anti-RNA polymerase I antibodies stained the same fibrillar structures in drug-segregated nucleoli as scleroderma sera. Scleroderma sera immunoprecipitated 13 polypeptides from (35S)methionine-labeled HeLa cell extract with molecular weights ranging from 210,000 to 14,000. Similar polypeptides were precipitated by rabbit anti-RNA polymerase I antibodies, and their common identities were confirmed in immunoabsorption experiments. Microinjection of purified IgG from a patient with speckled nucleolar staining effectively inhibited ribosomal RNA transcription. Autoantibodies to RNA polymerase I were restricted to certain patients with scleroderma and were not found in other autoimmune diseases.
Vasoactive agents which elevate either cGMP or cAMP inhibit platelet activation by pathways sharing at least one component, the 46/50 kDa vasodilator-stimulated phosphoprotein (V ASP). V ASP is stoichiometrically phosphorylated by both cGMP-dependent and cAMPdependent protein kinases in intact human platelets, and its phosphorylation correlates very well with platelet inhibition caused by cGMP- and cAMP-elevating agents. Here we report that in human platelets spread on glass, V ASP is associated predominantly with the distal parts of radial micro filament bundles and with microfilaments outlining the periphery, whereas less V ASP is associated with a central microfilamentous ring. V ASP is also detectable in a variety of different cell types including fibroblasts and epithelial cells. In fibroblasts, V ASP is concentrated at focal contact areas, along microfilament bundles (stress fibres) in a punctate pattern, in the periphery of protruding lamellae, and is phosphorylated by cGMP- and cAMP-dependent protein kinases in response to appropriate stimuli. Evidence for the direct binding of V ASP to F -actin is also presented. The data demonstrate that V ASP is a novel phosphoprotein associated with actin filaments and focal contact areas, i.e. transmembrane junctions between microfilaments and the extracellular matrix.
We have investigated Cd\(_{1-x}\)Mn\(_x\)Te thin films with Mn concentrations of x=0.12, 0.18, 0.30, 0.52, and 0.70. These single crystal layers were grown by molecular beam epitaxy on [001] CdTe substrates. The real part of the refractive index, n, was determined below the band-gap Eo in the range of 0.5-2.5 eV at T=300 K. The parallel reOectivity was measured near the Brewster angle at the YAG laser wavelength of 1.064 J.Lm (hv= 1.165 eV). Combining these results with the optical pathlength results (nd) of reOection measurements in a Fourier spectrometer we have determined n(x,v) over a wide spectral range by utilizing a three parameter fit. The accuracy of these results for n should improve waveguide designs based on this material.
Nuclei assembled in Xenopus egg extract from purified DNA or chromatin resemble their natural counterparts in a number of structural and functional features. However, the most obvious structural element of normal interphase nuclei, the nucleolus, is absent from the in vitro reconstituted nuclei. By EM, cytological silver staining, and immunofluorescence microscopy employing antibodies directed against various nucleolar components we show that nuclei assembled in vitro contain numerous distinct aggregates that resemble prenucleolar bodies (PNBs) by several criteria. Formation of these PNB-like structures requires pore complex-mediated nuclear transport of proteins but is independent of the genetic content of the in vitro nuclei as well as transcriptional and translational events. Our data indicate that nuclei assembled in vitro are capable of initiating early steps of nucleologenesis but that the resulting PNBs are unable to fuse with each other, probably due to the absence of a functional nucleolus organizer. With appropriate modifications, this experimental system should be useful to define and analyze conditions promoting the site-specific assembly of PNBs into a coherent nucleolar body.
Electron microscope preparations of lampbrush chromosomes from oocytes of Pleurodeles waltl;; have revealed a new class of tandemly repeated genes. These genes are highly active, as judged by the close spacing of nascent transcripts. They occur in clusters of >100 copies and are transcribed in units containing roughly 940 base pairs of DNA that are separated by nontranscribed spacers of an estimated DNA content of 2,410 base pairs. The size and the pattern of arrangement of these transcription units can not be correlated with any of the repetitious genes so far described.
A nucleolar skeleton of protein filaments demonstrated in amplified nucleoli of Xenopus laevis
(1981)
The amplified, extrachromosomal nucleoli of Xenopus oocytes contain a meshwork of -4-nm-thick filaments, which are densely coiled into higher-order fibrils of diameter 30-40 nm and are resistant to treatment with high- and low-salt concentrations, nucleases (DNase I, pancreatic RNase, micrococcal nuclease), sulfhydryl agents, and various nonionic detergents. This filamentous "skeleton" has been prepared from manually isolated nuclear contents and nucleoli as weil as from nucleoli isolated by fluorescence-activated particle sorting. The nucleolar skeletons are observed in light and electron microscopy and are characterized by ravels of filaments that are especially densely packed in the nucleolar cortex. DNA as weil as RNA are not constituents of this structure, and precursors to ribosomal RNAs are completely removed from the extraction-resistant filaments by treatment with high-salt buffer or RN ase. Fractions of isolated nucleolar skeletons show specific enrichment of an acidic major protein of 145,000 mol wt and an apparent pi value of -6.15, accompanied in some preparations by various amounts of minor proteins. The demonstration of this skeletal structure in "free" extrachromosomal nucleoli excludes the problem of contaminations by nonnucleolar material such as perinucleolar heterochromatin normally encountered in studies of nucleoli from somatic cells. It is suggested that this insoluble protein filament complex forms a skeleton specific to the nucleolus proper that is different from other extraction-resistant components of the nucleus such as matrix and lamina and is involved in the spatial organization of the nucleolar chromatin and its transcriptional products. In studies of the organization of the interphase nucleus, considerable progress has been made in the elucidation of the arrangement of chromatin components and transcriptional products. However, relatively little is known about the composition and function of another category of nuclear structures, the nonnucleoproteinaceous architectural components that are insoluble in solutions of low and high ionic strength, despite numerous studies dedicated to this problem. Such structures include (a) the nuclear envelope and its pore complexes (I, 15, 18, 23, 37, 41), (b) a peripheral layer of insoluble protein ("lamina"; I, 15, 22, 23, 59), (e) certain skeletal proteins related to the chromosome "scaffold" described by Laemmli and coworkers (see references 2 and 3), and (d) ill-defined tangles of fibrillar structures of the nuclear interior that are collectively described as residual "matrix" (6, 21 ; for reviews, see references THE JOURNAL OF CEll BrOlOGY . VOlUME 90 AUGUST 1981 289-299 © The RockefeIler University Press · 0021 -9525/ 81 / 08/ 0289/ 11 $1 .00 4 and 12). The latter, preparatively
Antibodies directed against RNA polymerase II (B) from Drosophila melanogaster were obtained from rabbit sera and, as monoclonal immunoglobulins, from mouse hybridomas and shown to cross-react with the amphibian enzyme protein. Localization by indirect immunofluorescence microscopy revealed the association of this enzyme with chromatin of interphase nuclei of amphibian cells and its absence in nucleoli. Purified immunoglobulins were microinjected in to nuclei ofliving vitellogenic oocytes of Ple1lrodeles waltlii and X enopus laevis and their effects on transcriptional processes were monitored by biochemical and light and electron microscopic stud ies. RNA polymerase II antibodies from rabbit sera caused a rapid and almost complete release of nascent transcripts from the chromatin axis of the loops of lampbrush chromosomes, followed by collapse of the loops and their retraction on the main chromosome axis. Monoclonal murine antibodies to the Iarge RNA polymerase II subunits also inhibited transcription in chromosome Ioops but appeared to inhibit initiation rather than elongation events. Activities of class land III RNA polymerases were not significantly affected by injection of antibodies to polymerase II, indicating immunological differences between the three RNA polymerases. The potential value of the in vitro test system described , as a very sensitive assay for detecting proteins involved in transcription in living cells, is discussed. 1
Human B cells appropriately activated by a B cell mitogen are rendered susceptible to human Interleukin 2 (IL-2) as demonstrated with recombinant human IL-2 (rec. h IL-2). They show increased proliferation and drastically enhanced immunoglobulin secretion. Susceptibility to IL-2 is accompanied with the expression of the IL-2 receptor (Tac antigen) on B cells. The data suggest that IL-2 is one of the lymphokines directly involved in the activation of B lymphocytes.
In this study the effect of recombinant human interleukin 2 (rec.hIL-2) on the proliferation and maturation of B lymphocytes was investigated. It was found that the presence of rec.hIL 2 results in proliferation of mitogen (LPS)-activated B cell blasts. In addition, it is shown that highly enriched murine B cells can be induced by rec.hIL-2 to proliferate and to develop into antibody-secreting cells (PFC) in the presence of antigen (SRBC). When tested for its effect on B cell preparations enriched for resting (small) or activated (blasted) B lymphocytes, it was found that rec.hIL 2 provides signals for both B cell populations to develop into PFC. In contrast, induction of proliferation by the same lymphokine source was only seen in blasted B cells. The data indicate that IL 2 is involved in the generation of B effector cells by directly acting on their precursors thereby providing differentiation as well as proliferation signals.
H-Y-specific and H-2Db-restricted, Lyt-1 "2+ T-cell clones (CTLL) with graded specific cytotoxic activities on male C57BL/6 (B6) target cells (1E3, +++; 2C5, ++; 2A5, +, 3E6, ±) were tested for their capacity to inhibit the generation of H-Y-specific cytotoxic T lymphocytes (CTL) in vitro. Addition of irradiated lymphocytes of CTLL 1E3 and CTLL 3E6 but not those of CTLL 2A5 or CTLL 2C5 abolished the generation of CTL from in vivo primed H-Y-specific precursor cells (CTLP) when added to fresh mixed-lymphocyte cultures (MLC). Exogenous sources of T-cell growth factors (TCGF) did not overcome suppression. Rather the presence of TCGF resulted in a further enhancement of suppressive activities in CTLL 1E3 and 3E6 and the induction of similar activities in cells from CTLL 2A5 and 2C5, which by themselves were not inhibitory. Moreover when added to similar MLC on Day 1 instead of Day 0, only irradiated cells of CTLL 3E6 but not those of the other three CTLL were suppressive. Induction of suppressive activities in H-Y-specific CTLL was independent of the appropriate male stimulator cells since it was also observed in MLC induced by irrelevant antigens (H-2, trinitrophenol). Furthermore at low cell numbers, irradiated lymphocytes from any of the CTLL consistently enhanced CTL activities generated from H-Y-specific CTLP. This augmenting activity, which was not TCGF, could be transferred by soluble mediators present in antigen-sensitized CTLL cultures. Thus, these data indicate (i) that cytotoxic effector cells can function as suppressor cells in the generation of CTL, (ii) that the cytotoxic activity of cloned CTL does not correlate with their capacity to suppress CTL responses, (iii) that the inhibition of CTL responses by CTLL is not due to simple consumption of T-cell growth factors produced in MLC, and (iv) that different CTL clones may interfere with the generation of CTL at different stages of their maturation. Moreover, the experiments suggest an antigen-independent enhancement of suppression by the interaction of CTL with lymphokines. Together with the augmenting activity evoked by cloned CTL the data provide strong evidence for the expression of multiple immunological functions by one particular subset of T cells and suggest that cytotoxic effector cells can differentially regulate the maturation and/or clonal expression of their precursor cells.
Mouse H-Y-specific and I-Ab restricted T-cell clones have been established and compared for their helper effects in the differentiation ofboth T and B Iymphocytes. The results demonstrate that three individual T -cell clones and one subclone could help in the antigen-driven induction of cytotoxic Iymphocytes (CTL) from their precursor cells (CTL-P), and were able to activate B cells to develop into antibody-secreting cells (PFC) in the presence of SRBC, provided the cloned T cells were restimulated by H-Y antigen on antigen-presenting cells. In addition, antigen or lectin could induce the same H -Y -specific T -cell clones to secrete factor(s) expressing helper activities similar to that ofthe cloned T cells. Furthermore, it is shown that the T cell-derived soluble mediator(s) was distinct from T-cell growth factor (TCGF) and from immune interferon (lFN-y). The data reveal a new type ofT cell with helper potential for the activation ofCTL-P and B Iymphocytes, and suggest the existence of distinct T helper cells which can provide help for both cytotoxic and antibody responses by virtue of different Iymphokine activities.
We have recently demonstrated that the frequency ofT cells expressing granzyme A is significantly higher in skin lesions and spleens of susceptible BALB/c mice compared with resistant C57BL/6 mice infected with Leishmania major, a cause of human cutaneous leishmaniasis. In the present study, we have performed in vitro studies to characterize the subpopulation, the antigen responsiveness and the lymphokine production pattern of granzyme A-expressing T cells in L. major-infected mice. Using a limiting dilution system for functional analysis of selected T cells at the clonallevel, we could show that granzyme A activity in infected BALB/c mice can be assigned to L. major-reactive CD4\(^+\) T cells secreting interleukin-2 (IL-2) and IL-4. Granzyme A production was most pronounced in the early phase of infection. On the other hand, granzyme A expression could not be detected in C57BL/6-derived T cells responding to L. major. The da ta support the suggestion that granzyme A is produced by L. major-responsive CD4\(^+\) T cells facilitating lesion formation and the dissemination of infection.
The pioneer tree Macaranga in SE Asia has developed manyfold associations with ants. The genus comprises all stages of interaction with ants, from facultative relationships to obligate myrmecophytes. Only myrmecophytic Macaranga offer nesting space for ants and are associated with a specific ant partner. The nonmyrmecophytic species are visited by a variety of different ant species which are attracted by extrafloral nectaries (EFN) and food bodies. Transitional Macaranga species like M. hosei are colonized later in their development due to their stem structure. Before the colonization by their specific Crematogaster partner the young plants are visited by different ant species attracted by EFN. These nectaries are reduced and food body production starts as soon as colonization becomes possible. We demonstrated earlier that obligate ant partners can protect their Macaranga plants against herbivore damage and vine cover. In this study we focused on nonspecific interactions and studied M. tanarius and M. hosei, representing a non-myrmecophyte and a transitional species respectively. In ant exclusion experiments both M. tanarius and M. hosei suffered significantly higher mean leaf damage than controls, 37% versus 6% in M. hosei, 16% versus 7% in M. tanarius. M. tanarius offers both EFN and food bodies so that tests for different effects of these two food rewards could be conducted. Plants with food bodies removed but with EFN remaining had the lowest mean increase of herbivore damage of all experimental groups. Main herbivores on M. hosei were mites and caterpillars. Many M. tanarius plants were infested by a shootborer. Both Macaranga species were visited by various ant species. Crematogaster spp. being the most abundant. We found no evidence for any specific relationships. The results of this study strongly support the hypothesis that non-specific, facultative associations with ants can be advantageous for Macaranga plants. Food bodies appear to have lower attractive value for opportunistic ants than EFN and may require a specific dietary adaptation. This is also indicated by the fact that food body production in the transitional M. hosei does not start before stem structure allows a colonization by the obligate Crematogaster species. M. hosei thus benefits from facultative association with a variety of ants until it produces its first domatia and can be colonized by its obligate mutualist.
C. borneensis (Myrmicinae) lives in dose association with several myrmecophytic species of the South East Asian pioneer tree genus Macaranga (Euphorbiaceae). The ants are adapted to the plants so dosely that they do not survive away from it. The only food they utilize is provided as food bodies by the plant and honeydew from specific scale insects kept inside the hollow internodes. The anatomy of the digestive tract is also adapted to life on the host plant: the crop is very sm all and can store only minute food quantities. C. borneensis exdusively colonizes certain Macaranga species. Queens as weIl as workers are able to recognize their host plant species, probably by chemical cues. Colony founding queens swarm throughout the year, mostly during darkness. There is strong competition among queens for host plants. Queens do not carry scale insects on their nuptial flight. Worker ants are active day and night. Most of them patrol and collect food bodies on the younger parts of the host plant. An important characteristic is their deaning behaviour, which results in removal of aIl foreign objects. Even though they are rather smalI, workers respond very aggressively to certain kinds of disturbance of the host plant. The ants attack most phytophagous insects and are especially effective in killing and removing smalI, softbodied herbivores (e.g. caterpillars). They do not possess a functional sting, but apply defensive secretion and-once biting an intruder-will not let go. Their effective alarm system results in a mass attack, which provides adequate defence for the colony and the host plant. A comparison with another Crematogaster species further illustrated the special adaptations of C. borneensis to its host plant.
At early developmental stages (embryonic day 7, E7), chick paravertebral sympathetic ganglia contain a cell population that divides in culture while expressing various neuronal properties. In an attempt to identify factors that control neuronal proliferation, we found that ciliary neurotrophic factor (CNTF) specifically inhibits the proliferation of those cells expressing neuronal markers. In addition, CNTF affects the differentiation of sympathetic ganglion cells by inducing the expression of vasoactive intestinal peptide immunoreactivity (VIP-IR). After 1 day in culture, tyrosine hydroxylase immunoreactivity (TH-I R) was expressed by about 86% of the cells whereas VIP-IR was virtually absent. In the presence of CNTF, 50%-60% of the cells expressed VIP-IR after 4 days in culture; however, none of the cells expressed VIP-IR in the absence of CNTF. These results, and the demonstration of cells that express both VIP and TH-IR, indicate that VIP is induced in cells that initially express tyrosine hydroxylase. The findings suggest a potential role for CNTF as a factor affecting the proliferation and differentiation of developing sympathetic neurons.
Motoneuron diseases represent a m&jor challenge to modern neurology, yet their clinical manifestations ware first described more than hundred years ago, and despite many studies the etiology of these diseases ramd,ns obscure with no effective treatments having been reported. Although progress has been made in establishing genetic linkage in the rare inherited for.ms of these diseases such as familial amyotrophic lateral scleriosisl , spinal mDscular atrophy and X-linked bulbo-spinal-mDscular atrophy, this new information has not yet affected therapeutic techniques. During the last few years several important steps have been taken concerning the physiological mechanisms involved in motoneuron survival during development, after lesion and in animal models of degenerative diseases, the molecular clOning of several new neurotrophic factors (brain-derived neurotrophic factor (BDNP), neurotrophin-3 and-4 (NT-3 and NT-4) and ciliary neurotrophic factor (CNTP)); the identification of a gene family of receptor molecules for same of these factors, progress in the understanding of the effects of polypeptide growth factors on muscle cell differentiation, neuronal sprouting (insulin-like growth factor-I and -11 (IGF-I and IGF-II), and in vitro motoneuronal survival (CNTF, IGF-I and -II and basic FGF). These findings have raised new hopes in that they could lead to a better understanding of the pathophysiological processes underlying these diseases, and that the pharmacological use of same of these newly characterized neurotrophic factors could present new possibilities for the treatment of these diseases.
An Metabasiten der Zone Erbendorf-Vohenstrauß, der Erbendorfer Grünschieferzone und der Zone Tirschenreuth-Mähring wurden petrographische, geochemische und phasenpetrologische Untersuchungen sowie K-Ar-Mineraldatierungen durchgeführt. In der Zone Erbendorf-Vohenstrauß treten in regionaler Abhängigkeit drei Haupttypen von Metabasiten auf: 1. Flaseramphibolite mit der Paragenese Hornblende + Oligoklas/ Andesin + Granat ± Salit und mit Plagioklas- und / oder Kalksilikat-reichen Flasern findet man hauptsächlich im nördlichen und zentralen Teil der Zone Erbendorf-Vohenstrauß. Geochemisch zeigen sie eine deutliche Anreicherung der inkompatiblen Elemente und der leichten Seltenerdelemente und sind vergleichbar mit modernen Tholeiiten von ozeanischen Inseln oder anomalen Abschnitten mittelozeanischer Rükken. Ein kontinentaler Intraplatten-Charakter scheint nicht gegeben. In der Kontaktaureole von Windisch-Eschenbach wurde ein Teil der Flaseramphibolite kontaktmetamorph überprägt. 2. Schiefrige, teils streifige Amphibolite mit der Paragenese Hornblende + Andesin / Labrador ± Salit dominieren im Süd teil der Zone Erbendorf-Vohenstrauß, erscheinen aber in einzelnen Ausstrichen auch im Norden. Die Streifung entsteht durch Plagioklas- und / oder Kalksilikat-reiche Zwischenlagen. Geochemisch sind diese Amphibolite bei flachen Spurenelement- und Seltenerdelement- Mustern vergleichbar mit modernen Tholeiiten von normalen Abschnitten mittelozeanischer Rücken. Sowohl Flaseramphibolite als auch schiefrige, teils streifige Amphibolite zeigen Übergänge zu massigen, homogenen Amphiboliten. 3. Metagabbros mit der Paragenese Hornblende + Plagioklas ± Biotit und einer grobkörnigen, gabbro iden Textur treten hauptsächlich im Bereich der Schuppenzone Michldorf-Kaimling (VOLL 1960) auf. Geochemisch nehmen sie bei einer schwachen Anreicherung der inkompatiblen Elemente eine Zwischenstellung zwischen Flaseramphiboliten und schiefrigen, teils streifigen Amphiboliten ein. In der Erbendorfer Grünschieferzone treten Metagabbros und schiefrige, teils gestreifte Amphibolite mit der Paragenese Hornblende + Oligoklas ± Epidot ± Chlorit auf. Die Amphibolite wurden z. T. stark retrograd überprägt und dabei auch deformiert. Dies wird auf größere, post-regionalmetamorphe, tektonische Bewegungen, möglicherweise Deckenüberschiebungen zurückgeführt. Geochemisch sind die Metabasite der Erbendorfer Grünschieferzone mit modernen tholeiitischen bis kalkalkalischen, subduktionsgebundenen Basalten vergleichbar. Sie unterscheiden sich damit völlig von den südlich angrenzenden Flaseramphiboliten der Zone Erbendorf-Vohenstrauß und den nördlich benachbarten Amphiboliten des Fichtelgebirges. Dies unterstreicht die Stellung der Erbendorfer Grünschieferzone als eigenständige tektonische Einheit. Im Vergleich mit dem Münchberger Komplex zeigt die Erbendorfer Grünschieferzone große Ähnlichkeit mit der Prasinit-Phyllit-Serie hinsichtlich der tektonischen Stellung, der Lithologie und insbesondere des geochemischen Charakters der Metabasite. Zwischen der Zone Erbendorf-Vohen-strauß und den Münchberger Serien konnten bezüglich der Metabasite dagegen keine Parallelen festgestellt werden. Schiefrige, teils streifige Amphibolite der Zone Tirschenreuth-Mähring mit der Paragenese Hornblende + Andesin ± Salit (± Granat) und hellen Plagioklas- oder Kalksilikat-reichen Lagen sind geochemisch mit modernen Basalten normaler mittel ozeanischer Rücken vergleichbar. Die untersuchten Metabasite dürften die Zusammensetzung des basaltischen Ausgangsmaterials zumindest annähernd wiedergeben. In einigen Fällen wurde jedoch eine sekundäre Mobilisation von Elementen festgestellt: Die kontaktmetamorphe Überprägung eines Teils der Flaseramphibolite in der Kontaktaureole von Windisch-Eschenbach führte zu einer deutlichen Anreicherung von Rb, Li und K und zu einer Abreicherung von Ca, Sr und V, insbesondere aber von Cr und Ni. Die sog. immobilen Elemente Nb, Ce, (P), Zr, Ti, Y, Sc streuen in einem deutlich breiteren Bereich als bei den nicht kontaktmetamorphen Flaseramphiboliten. In einer Probe sind die Seltenerdelemente insgesamt angereichert. Eine Mobilisation von P20 S und den leichten Seltenerdelementen in einigen schiefrigen, teils streifigen Amphiboliten der Zone Erbendorf-Vohenstrauß und in einigen kontaktmetamorphen Flaseramphiboliten wird auf einen Abbau von Apatit im Zuge einer post-regionalmetamorphen, möglicherweise postgranitischen, hydrothermalen Überprägung zurückgeführt. Mineralchemische Untersuchungen ergaben für die Zusammensetzung der Hornblenden aus Amphiboliten der Zone Erbendorf-Vohenstrauß deutliche regionale Unterschiede, die weitgehend unabhängig von der regionalen Verteilung der einzelnen Metabasit-Haupttypen sind. Vergleichende Untersuchungen an Granat untermauern eine vermutete, frühere, eklogitfazielle Überprägung eines Granatamphibolits vom Nordrand der Zone Erbendorf-Vohenstrauß bei Hauxdorf. K-Ar-Altersdatierungen an Hornblende- und Glimmerkonzentraten belegen, daß das letzte, (Mitteldruck-) metamorphe Ereignis im Westteil der Zone Erbendorf-Vohenstrauß vor etwa 380 Ma beendet war. Dieses Alter wird korreliert mit einer letzten Metamorphose der höhermetamorphen Serien des Münchberger Komplexes zur gleichen Zeit. Im Ostteil der Zone Erbendorf-Vohenstrauß treten jüngere Altersdaten mit einem Schwerpunkt bei 324 Ma auf, die entweder auf einen Einfluß der etwa gleichalten Niederdruck-Metamorphose im unmittelbar benachbarten Moldanubikum i. e. S. oder auf die Intrusion des 324 Ma alten (KÖHLER et al. 1974) Leuchtenberger Granits zurückzuführen sind. Im Übergangsbereich vom Saxothuringikum zum Moldanubikum bei Mähring zeigen Altersdaten um 320Ma das Ende der letzten (Niederdruck-)Metamorphose an.
Nonnucleolar chromatin from interphase nuclei of Physarum polycephalum plasmodia occurs in two different structural configurations as seen in electron microscopic spread preparations. While the majority of the chromatin is devoid of nascent ribonucleoprotein (RNP) fibrils and compacted into nucleosomal particles, a minor proportion (10- 20%) is organized differently and reveals a smooth contour. It is this form of smooth chromatin which is rich in transcription units (mean length: 1.36±0.21 11m). Only occasionally are solitary nascent RNP fibrils observed which are associated with beaded strands of chromatin. In transcribed smooth chromatin nucleosomal particles are not only absent from the transcription units but also from their nontranscribed flan king regions, indicating that this special structural aspect is not merely a direct consequence of the transcriptional process. The existence of ca. 10- 20% of Physarum chromatin in the smoothly contoured form is discussed in relation to reports of a preferential digestibility of a similar proportion of Physarum chromatin by DNAse I (Jalouzot et al. , 1980) and to the altered configuration of "peak A" chromatin subunits after micrococcal nuclease digestion (Johnson et al., 1978a, b).
Oocytes of the water beetle, Dytiscus marginalis, contain large amounts of rDNA most of which is present in the form of rings containing one or several pre-rRNA genes. Electron microscopy of spread preparations of vitellogenic oocytes has shown that the rDNA is extended in chromatin rings with transcribed pre- rRNA genes and is not packed into nucleosomes (Trendelenburg eta!. , 1976). When similar preparations are made from previtellogenic ooytes in which a large proportion of the nuc1eolar chromatin is transcriptionally inactive, a different morphological form of this chromatin is recognized. In contrast to the transcribed chromatin rings the inactive nucleolar chromatin circles show the characteristic beaded configuration, indicative of nucleosomal packing. Nuc1eosomal packing is also indicated by the comparison of the lengths of these chromatin rings with both iso lated rDNA circ1es and transcribed chromatin rings. In addition, these inactive nuc1eofilaments often appear to be compacted into globular higher order structures of diameters from 21 to 34nm, each composed of an aggregate of 6-9 nuc1eosomes. While the estimated reduction of the overall length of rDNA, as seen in our preparations, is, on the average, only 2.2 - 2.4 fold in the nuc1eosomal state it is 10- 13 fold when supranuc1eosomal globules are present. These data show that the extrachromosomal rDNA of these oocytes goes through a cycle of condensation and extensio n, as a function of the specific transcriptional activity, and that the beaded state described here is exc1usively found in the non-transcribed state.
Lengths and patterns of transcriptional units in the amplified nucleoli of oocytes of Xenopus laevis
(1977)
Transcriptionally active chromatin from peripheral amplified nuc1eoli of lampbrush-chromosome stage oocytes of Xenopus laevis was dispersed and spread in various solutions of low salt concentrations (incIuding some with additions of detergents) and examined by electron microscopy. Nucleolar material from oocytes of animals with normal (2-nu) and mutant (I-nu) genetical constitution of nucleolus organizers was compared. Histograms showing the distributions of the lengths of matrix units, apparent spacer intercepts, and the total repeating units of the rDNA containing chromatin axes revealed a significant degree of heterogeneity, with indications of subclasses and predominant repeat unit size c1asses of 3.3 and 3.8 11m length. The correspondence of matrix unit length to the molecular weight of the first stable product of rDNA transcription was studied using gel electrophoresis of labelIed pre-rRNA under non-denaturing and denaturing conditions. Evaluations of individual strands of nucleolar chromatin furt her demonstrated the existence of both (i) strands with obviously homogeneous repeating units and (ii) strands with intra-axial heterogeneity of rDNA subunits. " Preludecomplexes ", i.e. groups of transcriptional complexes in apparent spacer intercepts, were not infrequently noted. The data are compared with the measurements of lengths of repeating units in fragments of rDNA obtained by digestion with EcoRI endonuclease as described by Morrow et al. (1974) and Wellauer et al. (1974, 1976a, b). The results are discussed in relation to problems of variations in the modes of arrangement of the pre-rRNA genes, the state of packing of rDNA during transcription, and possible mechanisms of the amplification process.
The Ringgold Knoll pegmatite, a late-stage member of the Granite Harbour Intrusives, crosscuts high-grade Wilson gneisses of the Oates Coast, which forms the westernmost part of the Wilson Terrane at the Pacific end of the Cambro-Ordovician Ross orogenic belt in West Antarctica. The pegmatite mineral assemblage consists of K-feldspar, plagioclase, quartz, garnet (almandinespessartine-pyrope), dark tourmaline (schorl-dravite), muscovite, apatite, monazite, zircon, blue AI-rich tourmaline and dumortierite in order of decreasing abundances. Major, minor and rare earth elements are reported for the greater part of the mineral assemblage. The time of pegmatite emplacement is constrained by Rb-Sr and Sm-Nd isochron ages of 492 ± 8 (2a) Ma and 500 ± 40 (2a) Ma, respectively. High initial 87Sr/86Sr of 0.7315 ± 0.0003 and low E Nd,t of -8.7 ± 1.2 strongly support an origin of the magma from highly evolved crustal source rocks. K-Ar and Ar-Ar model ages of about 470 to 475 Ma for igneous muscovite indicate that the pegmatite together with its wall rocks spent a prolonged period at elevated temperatures before final cooling below about 350 °C. The muscovite dates may give an estimate for the time of exhumation of the Oates Coast crystalline basement along two major late Ross orogenic detachment zones within the Wilson Terrane i.e. the Wilson and the Exiles thrusts (c.f. FLÖTTMANN and KLEINSCHMIDT, 1991).
The paleotropical tree genus Macaranga (Euphorbiaceae) comprises all stages of interaction with ants, from facultative associations to obligate myrmecophytes. In SE.-Asia food availability does not seem to be the limiting factor for the development of a close relationship since all species provide food for ants in form of extrafloral nectar and/or food bodies. Only myrmecophytic Macaranga species offer nesting space for ants (domatia) inside intern odes which become hollow due to degeneration of the pith. Non-myrmecophytic species have a solid stem with a compact and wet pith and many resin ducts. The stem interior of some transitional species remains solid, but the soft pith can be excavated. The role of different ant-attracting attributes for the development of obligate ant-plant interactions is discussed. In the genus Macaranga, the provision of nesting space seems to be the most important factor for the evolution of obligate myrmecophytism.
Nuclear envelopes of maturing oocytes of various amphibia contain an unusually high number of pore complexes in very close packing. Consequently, nuclear envelopes , which can be manually isolated in great purity, provide a remarkable enrichment of nuclear pore complex material, relative to membranous and other interporous structures. When the polypeptides of nuclear envelopes isolated from oocytes of Xenopl/s la evis and Triturus alpestris are examined by gel electrophoresis, visualized either by staining with Coomassie blue or by radiotluorography after in vitro reaction with [3H]dansyl chloride , a characteristic pattern is obtained (10 major and 15 minor bands). This polypeptide pattern is radically different from that of the nuclear contents isolated from the same cell. Extraction of the nuclear envelope with high salt concentrations and moderateIy ac tive detergents such as Triton X- 100 results in the removal of membrane material but leaves most of the non-membranous structure of the pore complexes. The dry weight of the pore complex (about 0.2 femtograms) remains essentially unchanged during such extractions as measured by quantitative electron microscopy . The extracted preparations which are highly enriched in nuclear pore complex material contain only two major polypeptide components with apparent molecular weights of 150000 and 73000. Components of such an electrophoretic mobility are not present as major bands , if at all , in nuclear contents extracted in the same way. lt is concluded that these two polypeptides are the major constituent protein(s) of the oocyte nuclear pore complex and are specific for this structure. When nuclear envelopes are isolated from rat liver and extracted with high salt buffers and Triton X- 100 similar bands are predominant, but two additional major components of molecular weights of 78000 and 66000 are also recognized. When the rat liver nuclear membranes are further subfractionated material enriched in the 66000 molecular weight component can be separated from the membrane material, indicating that this is relatively loosely associated material , probably a part of the nuclear matrix . The results suggest that the nuclear pore complex is not only a characteristic ubiquitous structure but also contains similar, if not identical , skeletal proteins that are remarkably re sistant to drastic changes of ionic strength as weil as to treatments with detergents and thiol reagents.
Sizes of chromosome loops and hnRNA molecules in oocytes of amphibia of different genome sizes
(1982)
The lengths of lampbrush chromosome loops and their tran scription units show a positive correlation with genome size in oocytes of amphibia with different C values. However, there is no such correlation with contour lengths of hnRNA molecu les isolated from these oocytes. These results indi cate th at more ON A sequences are transcribed in amphibia of higher C value , but that processing of RNA transc ripts occurs while they are still attached to the chromosomes as nascent ribonucleoprotein fibrils.
Under the intluence of 5-tluoro-uridine, the ultrastructure of the rDNA transcription units in Xenopus oocytes is altered. Whereas part of the matrix units maintains anormal aspect or shows various degrees of inhibition, in a strong proportion of the transcription units the alternating pattern of matrix units and fibril-free spacer regions is no longer recognized. Transcriptional complexes are found along the entire DNP axis, including the regions of the spacers. These observations support biochemical data on transcription in rDNA spacer region.
Transcriptionally inactive chick erythrocyte nudei were reactivated by Sendai virusinduced fusion of erythrocytes with rat L6j1 myoblasts. We used antibodies to trace the appearance of a specific protein engaged in transcription of a defined dass of genes, those coding for rRNA, during reactivation. Using immunofluorescence microscopy, we found increasing amounts of rat RNA polymerase I to appear, during a certain period of time after fusion, in the reforming nudeoli of the chick nudei. Amounts of rat RNA polymerase I sufficient to be detected by immunofluorescence microscopy had accumulated in the newly developed chick nudeoli 72- 190 h after fusion was initiated. This time interval coincides with the time when chick rRNA synthesis can first be detected. The results raise the possibility that during these stages of the reactivation process chick rRNA genes are transcribed by heterologous RNA polymerase I moleeules of rat origin.
Upon incubation of cultured rat cells with the adenosine analogue 5,6-dichloro-l-β- D-ribofuranosylbenzimidazole (DRB), nucleoli reversibly dissociate into their substructures, disperse throughout the nuclear interior, and form nucleolar "necklaces". We have used this experimental system, which does not inhibit transcription of the rRNA genes, to study by immunocytochemistry the distribution of active rRNA genes and their transcriptional products during nucleolar dispersal and recovery to normal morphology. Antibodies to RNA polymerase I allow detection of template-engaged polymerase, and monoclonal antibodies to a ribosomal protein (S 1) of the small ribosomal subunit permit localization of nucleolar preribosomal particles. The results show that, under the action of DRB transcribed rRNA, genes spread throughout the nucleoplasm and finally appear in the form of several rows, each containing several (up to 30) granules positive for RNA polymerase land argyrophilic proteins. Nucleolar material containing preribosomal particles also appears in granular structures spread over the nucleoplasm but its distribution is distinct from that of rRNA gene-containing granules. We conclude that, although transcriptional units and preribosomal particles are both redistributed in response to DRB, these entities retain their individuality as functionally defined subunits. We further propose that each RNA polymerase-positive granular unit represents a single transcription unit and that each continuous array of granules ("string of nucleolar beads") reflects the linear distribution of rRNA genes along a nucleolar organizer region. Based on the total number of polymerase I-positive granules we estimate that a minimum of 60 rRNA genes are active during interphase of DRB-treated rat cells.
Purified mitochondrial DNA (mitDNA) from ovaries ofXenopus lae vis was injected into the nuclei (germinal vesicles) of large viteUogenic oocytes of the same organism and examined by electron microscopy ofthe spread nuclear contents. Normally located nuclei of untreated oocytes as weil as peripherally translocated nuclei of centrifuged oocytes were used. In addition, oocyte nuclei isolated and incubated under liquid paraffin oil were injected with DNA. The integrity oftranscriptional structures of endogenous chromosomal (Iampbrush chromosomes) and extrachromosomal (nucleoli) genes of the injected nuclei was demonstrated. Microinjected mitDN A was identified as circles of chromatin exhibiting polynucleosome-like organization and a me an contour length of 2.6 J.Lm, corresponding to a compaction ratio of the mitDN A of about 2 : I. This DNA packing ratio is similar to that observed after preparation of various kinds of native chromatin in low salt buffers. The chromatin circles formed from injected mitDNA only very rarely exhibited lateral fibrils suggestive of transcriptional activity. These results suggest that purified mitDNA can be transformed to normally structured chromatin when exposed to oocyte nuclear contents but is rarely , if at all , transcribed in this form and in this environment.
The structure of the rat ciliary neurotrophic factor (CNTF) gene and the regulation ofCNTF mRNA levels in cultured glial cells were investigated. The rat mRNA is encoded by a simple two-exon transcription unit. Sequence analysis of the region upstream of the transcription start-site did not reveal a typical TATA-box consensus sequence. Low levels of CNTF mRNA were detected in cultured Schwann cells, and CNTF mRNA was not increased by a variety of treatments. Three-week-old astrocyteenriched cell cultures from new-born rat brain contained easily detectable CNTF mRNA. In astrocyte-enriched cultures, upregulation of CNTF mRNA levels was observed after treatment with IFN-gamma. CNTF mRNA levels were down-regulated in these cells by treatments that elevate intracellular cyclic AMP and by members of the fibroblast growth factor (FGF) family. The implications of these results for potential in vivo functions of CNTF are discussed.
Ciliary neurotrophic factor (CNTF) is expressed in high quantities in Schwann cells of peripheral nerves during postnatal development of the rat. The absence of a hydrophobic leader sequence and the immunohistochemical localization of CNTF within the cytoplasm of these cells indicate that the factor might not be available to responsive neurons under physiological conditions. However, CNTF supports the survival of a variety of embryonic neurons, including spinal motoneurons in culture. Moreover we have recently demonstrated that the exogenous application of CNTF protein to the lesioned facial nerve of the newborn rat rescued these motoneurons from cell death. These results indicate that CNTF might indeed play a major role in assisting the survival of lesioned neurons in the adult peripheral nervous system. Here we demonstrate that the CNTF mRNA and protein levels and the manner in which they are regulated are compatible with such a function in lesioned peripheral neurons. In particular, immunohistochemical analysis showed significant quantities of CNTF at extracellular sites after sciatic nerve lesion. Western blots and determination of CNTF biological activity of the same nerve segments indicate that extracellular CNTF seems to be biologically active. After nerve lesion CNTF mRNA levels were reduced to <5 % in distal regions of the sciatic nerve whereas CNTF bioactivity decreased to only one third of the original before-lesion levels. A gradual reincrease in Schwann cells occurred concomitant with regeneration.