On the evolution of protamines in bony Fish: Alternatives to the “Retroviral horizontal transmission” hypothesis

Fish protamines are highly specialized molecules which are responsible for chromatin condensation during the last stages of spermatogenesis (spermiogenesis). However, not all fish contain protamines in their sperm nuclei; rather, there seems to be a random distribution of protamines within this group. The origin of this sporadic presence of protamines in the sperm and its significance have not yet been precisely determined. In this paper we have conducted an exhaustive survey of the literature available on the different types of nuclear protein composition of the sperm of teleost fish in order to try to correlate these data with what is presently known about the taxonomy of this group. The results of this analysis have allowed us to make the following observations. The divergence between protamines and histones has occurred several times during the evolution of the bony fish. However, the relative frequency of this divergence is almost negligible during the differentiation of genera and species (intrafamily variation) and is very small during the differentiation of families (interfamily variation). Nevertheless, the divergence is very noticeable among the different orders. It is therefore possible to conclude from all this that the sporadic distribution of protamines in bony fish is not a random event as initially believed. Furthermore, such a heterogeneous distribution of protamines cannot be easily accounted for by a mechanism of horizontal retroviral transmission through repeated and independent acquisition of a prot amine gene as has been recently proposed (Jankowski, Stater, Dixon (1986) J Mol Evol 23:1–10). Rather, it could possibly be explained by a repeated and independent loss of the expression of the protamine gene (or loss of the gene itself) which mainly occurred during the diversification of the orders of this group.Correspondence to: J. Ausio     

New species and a new combination in Myrtaceae from northeastern South America

Three new species of Myrtaceae (Calyptranthes bracteata, Eugenia gonglycocarpa, andMyrcia rupta) from northeastern South America are described and illustrated, and a new combination (Eugenia tetramera) is proposed. The closed-calyx and the completely or partially fused cotyledons ofMyrcia rupta, unusual features for the genus, are discussed and compared with related species inMyrcia andMarlierea.     

Crop/Weed gene flow:Cucurbita argyrosperma Huber andC. fraterna L. H. Bailey (Cucurbitaceae)

A mixed population of Cucurbita at Vado El Mow in northern Tamaulipas, Mexico showed an anomalous pattern of fruit bitterness. Some domesticated plants (C. argyrosperma andC. moschata) expressed cucurbitacin bitterness whereas some sympatric free-living plants produced non-bitter fruits. This reversal of typical cucurbitacin expression suggested gene flow between crop and weed at the site. Isozyme analysis provided little insight as to taxa involved in gene exchange, although progeny from a single free-living plant carried IDH allozymes that are associated with Mexican landraces ofC. pepo. Synthetic hybridization revealed that fertile F, hybrids are produced from crosses involvingC. fraterna as the pistillate parent andC. argyrosperma as the staminate parent. Interspecific crop/weed hybrids can produce viable progeny upon self-pollination or backcrossingto either parent, andF₂ families display normal allozyme segregation. Hybrid fertility, as indicated by pollen stainability, increases in progeny produced by backcrossingfrom theC. argyrosperma parent. Interspecific hybridfertility represents a potential for crop/weed gene flow that would be realized under natural conditions if pollen flow occurs betweenC. fraterna andC. argyrosperma in the fields of Tamaulipas. Oligolectic “squash bees” (Teponapis), efficientCucurbita pollen vectors, are present at the site. Thus, it is likely that natural interspecific crop/weed hybridization has occurred at Vado El Moro and this might at least partially explain the anomalous distribution of fruit bitterness among extant populations at the site.     

Stabilité du polymorphisme enzymatique dans les populations d''un Lépidoptère migrant,Agrotis ipsilon

Homogeneous population structure in a migrant Lepidoptera, Agrotis ipsilon. Light trapping of Agrotis ipsilon (Lepidoptera, Noctuidae) on various passes of the Alps and Pyrénées exhibited wide range movements between overwintering and aestivation areas. Electrophoretic analysis of samples taken in the Cantons of Vaud and Tessin (Switzerland), in the Rhône Delta (Southern France), and on passes of the Alps and Pyrénées, showed a great temporal and spatial homogeneity of allele frequencies (Fst values ranging from 0.002 to 0.013, and genetic distances from 0 to 0.004). These results support the hypothesis of a high level of gene flow. However, the occurrence during some years of high Fis values, might be explained by mixtures of populations that had undergone selection or went through a bottle-neck.     

Compositional heterogeneity of the Escherichia coli genome: A role for VSP repair?

E. coli genes that contain a high frequency of the tetranucleotide CTAG are also rich in the tetramers CTTG, CCTA, CCAA, TTGG, TAGG, and CAAG (group-I tetramers). Conversely, E. coli genes lacking CTAG are rich in the tetranucleotides CCTG, CCAG, CTGG, and CAGG (group-II tetramers). These two gene samples differ also in codon usage, amino acid composition, frequency of Dcm sites, and contrast vocabularies. Group-I tetramers have in common that they are depleted by very-short-patch repair (VSP), while group-II tetramers are favored by VSP activity. The VSP system repairs G:T mismatches to G:C, thereby increasing the overall G+C content of the genome; for this reason the CTAG-rich sample has a lower G+C content than the CTAG-poor sample. This compositional heterogeneity can be tentatively explained by a low level of VSP activity on the CTAG-rich sample. A negative correlation is found between the frequency of group-I tetramers and the level of gene expression, as measured by the Codon Adaptation Index (CAI). A possible link between the rate of VSP activity and the level of gene expression is considered.Correspondence to: A. Marine     

Brain ATP Depletion Induced by Acute Ammonia Intoxication in Rats Is Mediated by Activation of the NMDA Receptor and Na+, K+-ATPase

Abstract: Injection of large doses of ammonia into rats leads to depletion of brain ATP. However, the molecular mechanism leading to ATP depletion is not clear. The aim of the present work was to assess whether ammonium-induced depletion of ATP is mediated by activation of the NMDA receptor. It is shown that injection of MK-801, an antagonist of the NMDA receptor, prevented ammonia-induced ATP depletion but did not prevent changes in glutamine, glutamate, glycogen, glucose, and ketone bodies. Ammonia injection increased Na⁺,K⁺-ATPase activity by 76%. This increase was also prevented by previous injection of MK-801. The molecular mechanism leading to activation of the ATPase was further studied. Na⁺,K⁺-ATPase activity in samples from ammonia-injected rats was normalized by "in vitro" incubation with phorbol 12-myristate 13-acetate, an activator of protein kinase C. The results obtained suggest that ammonia-induced ATP depletion is mediated by activation of the NMDA receptor, which results in decreased protein kinase C-mediated phosphorylation of Na⁺,K⁺-ATPase and, therefore, increased activity of the ATPase and increased consumption of ATP.     

Asymmetric somatic hybridization between tall fescue (Festuca arundinacea Schreb.) and irradiated Italian ryegrass (Lolium multiflorum Lam.) protoplasts

Intergeneric asymmetric somatic hybrids have been obtained by the fusion of metabolically inactivated protoplasts from embryogenic suspension cultures ofFestuca arundinacea (recipient) and protoplasts from a non-morphogenic cell suspension ofLolium multiflorum (donor) irradiated with 10, 25, 50, 100, 250 and 500 Gy of X-rays. Regenerating calli led to the recovery of genotypically and phenotypically different asymmetric somatic hybridFestulolium plants. The genome composition of the asymmetric somatic hybrid clones was characterized by quantitative dot-blot hybridizations using dispersed repetitive DNA sequences specific to tall fescue and Italian ryegrass. Data from dot-blot hybridizations using two cloned Italian ryegrass-specific sequences as probes showed that irradiation favoured a unidirectional elimination of most or part of the donor chromosomes in asymmetric somatic hybrid clones obtained from fusion experiments using donor protoplasts irradiated at doses 250 Gy. Irradiation of cells of the donor parent with 500 Gy prior to protoplast fusion produced highly asymmetric nuclear hybrids with over 80% elimination of the donor genome as well as clones showing a complete loss of donor chromosomes. Further information on the degree of asymmetry in regenerated hybrid plants was obtained from chromosomal analysis including in situ hybridizations withL. multiflorum-specific repetitive sequences. A Southern blot hybridization analysis using one chloroplast and six mitochondrial-specific probes revealed preferentially recipient-type organelles in asymmetric somatic hybrid clones obtained from fusion experiments with donor protoplasts irradiated with doses higher than 100 Gy. It is concluded that the irradiation of donor cells before fusion at different doses can be used for producing both nuclear hybrids with limited donor DNA elimination or highly asymmetric nuclear hybrid plants in an intergeneric graminaceous combination. For a wide range of radiation doses tested (25–250Gy), the degree of the species-specific genome elimination from the irradiated partner seems not to be dose dependent. A bias towards recipient-type organelles was apparent when extensive donor nuclear genome elimination occurred.Abbreviations cpDNA Chloroplast DNA - 2, 4-D 2,4-dichlorophenoxyacetic acid - FDA fluorescein diacetate - IOA iodoacetamide - mtDNA mitochondrial DNA - RFLP restriction fragment length polymorphism     

A population analysis of Robertsonian and Ag-NOR polymorphisms in brown trout (Salmo trutta)

An analysis of Robertsonian polymorphism and variation in the number of active NORs has been carried out in several populations of brown trout (Salmo trutta) from Northwestern Spain. The karyotype of this species appears to be soundly established, and essentially no variation has been found in chromosome number. Interindividual and interpopulation variation in arm number was detected, with figures ranging between 100 and 102 among individuals, and between 100.10 and 100.80 among populations. This variation in arm number is solely attributable to the polymorphism of the short arm of the main NOR-bearing pair 11, which can appear from acrocentric to metacentric in different individuals. Most populations analyzed showed the standard distribution of active NORs previously observed in this species. The Miño drainage basin, and specially the Chamoso population, showed a multi-chromosomal distribution of active NORs, with several new locations, always telomeric. In most cases no concordance was observed between previously detected rDNA sites in S. trutta and the new Ag-NOR locations. This fact suggests a transposition mechanism rather than an activation of silent rDNA sites to explain this multichromosomal NOR pattern.     

Rye chromosome arm 3RS encodes a homodimeric inhibitor of insect α-amylase

A new inhibitor of insect -amylase, designated RDAI-1, has been purified from rye (Secale cereale L.) endosperm. RDAI-1 is homologous to wheat homodimeric inhibitors. This homology is supported by their similar N-terminal amino-acid sequences, inhibitory activities towards amylases from Tenebrio molitor (Coleoptera) and human saliva, and aggregative properties in gel-filtration chromatography. The gene encoding RDAI-1, IdhaR1, is located on the short arm of chromosome 3R, which is homoeologous with wheat chromosome arms 3BS and 3DS, where the genes for homodimeric inhibitors have been previously mapped.     

Comparison of superovulatory response of mature outbred mice treated with FSH or PMSG and developmental potential of embryos produced

A superovulatory treatment for mice based on FSH administration was compared with a standard one based on PMSG. Our aim was to determine if a mean number of embryos recovered per donor could be increased and if in vitro or in vivo viability was affected by the hormonal treatment used. Thus, female Swiss mice were subjected to 2 superovulatory treatments, and the 1-cell and 2-cell stage embryos were cultured in 2 different media to the blastocyst stage or were transferred to pseudopregnant recipients. The data show that despite a lower mating percentage (52% with FSH vs 66% with PMSG), the FSH-treated mice provided twice the number of total embryos (53.4 vs 24.5) with a similar percentage of morphologically normal embryos (74% for FSH vs 69% for PMSG). We also found that in vitro culture results can be influenced by the source of gonadotropins depending on the culture medium used. A culture medium such as CZB which prevents the 2-cell block, provided the same developmental rates regardless of hormonal treatment used. However, with M-16 medium, which does not prevent this blockage, only 39% of the 2-cell FSH-derived embryos and 49% of the PMSG-derived 2-cell embryos developed into blastocysts (P<0.05). FSH-derived embryos resulted in a higher percentage of pregnant recipients (73 vs 56%) than PMSG-derived embryos, but the number of alive fetuses and the number of implantations per pregnant recipient was affected only by the kind of culture system used before transfer. The results show that FSH can provide very good superovulatory response in mice, thus reducing the number of donors needed for a given experiment and providing embryos of at least the same quality as those derived from the standard PMSG treatment.