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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.
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.
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.
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.