20 research outputs found
Time-course analysis of nuclear events during conjugation in the marine ciliate Euplotes vannus and comparison with other ciliates (Protozoa, Ciliophora)
No full textCiliates represent a morphologically and genetically distinct group of single-celled eukaryotes that segregate germline and somatic functions into two types of nuclei and exhibit complex cytogenetic events during the sexual process of conjugation, which is under the control of the so-called ‘mating type systems’. Studying conjugation in ciliates may provide insight lead to major advances into our understanding of the origins and evolution of sex and fertilization. In the present work, we studied in detail the sexual process of conjugation using the model species Euplotes vannus, and compared these nuclear events with those occurring in other ciliates. Our results indicate that in E. vannus: 1) conjugation requires about 75 hours to complete: the longest step is the development of the new macronucleus (ca. 64h), followed by the nuclear division of meiosis I (5h); the mitotic divisions usually take only 2h; 2) there are three prezygotic divisions (mitosis and meiosis I and II), and two of the eight resulting nuclei become pronuclei; 3) after the exchange and fusion of the pronuclei, two postzygotic divisions occur; two of the four products differentiate into the new micronucleus and macronucleus, respectively, and the parental macronucleus degenerates completely; 4) comparison of the nuclear events during conjugation in different ciliates reveals that there are generally three prezygotic divisions while the number of postzygotic divisions is highly variable. These results can serve as reference to investigate the mating type system operating in this species and to analyze genes involved in the different steps of the sexual process
Homo- and hetero-oligomeric protein–protein associations explain autocrine and heterologous pheromone-cell interactions in Euplotes
No full textIn Euplotes, protein pheromones regulate cell reproduction and mating by binding cells in autocrine or heterologous fashion, respectively. Pheromone binding sites (receptors) are identified with membrane-bound pheromone isoforms determined by the same genes specifying the soluble forms, establishing a structural equivalence in each cell type between the two twin proteins. Based on this equivalence, autocrine and heterologous pheromone/receptor interactions were investigated analyzing how native molecules of pheromones Er-1 and Er-13, distinctive of mating compatible E. raikovi cell types, associate into crystals. Er-1 and Er-13 crystals are equally ormed by molecules that associate cooperatively into oligomeric chains rigorously taking a mutually opposite orientation, and each burying two interfaces. A minor interface is pheromone-specific, while a major one is common in Er-1 and Er-13 crystals. A close structural inspection of this interface suggests that it may be used by Er-1 and Er-13 to associate into heterodimers, yet inapt to further associate into higher complexes. Pheromonemolecule homo-oligomerization into chains accounts for clustering and internalization of autocrine pheromone/receptor complexes in growing cells, while the heterodimer unsuitability to oligomerize may explain why heterologous pheromone/receptor complexes fail clustering and internalization. Remaining on the cell surface, they are credited with a key role in cell–cell mating adhesion
Morphology and molecular phylogeny of two new Aspidisca species (Ciliophora, Spirotrichea, Euplotida) collected from subtropical coastal waters in China
No full textCiliates are morphologically diverse and highly specialized unicellular eukaryotes that constitute an important component of the microbial food web. Aspidisca Ehrenberg, 1830, is a highly speciose genus that plays key ecological roles in a wide range of environments, and its species diversity has always been a hot spot in protozoan research. In this study, we investigate the living morphology, infraciliature, silverline system, and ribosomal small subunit (SSU rRNA) gene sequences of two new Aspidisca species collected from subtropical coastal waters of China using standard methods. Aspidisca spina sp. n. is characterized by having an obvious peristomial spur, two transparent posterior protrusions, seven frontoventral cirri in “polystyla-arrangement”, and six dorsal ridges. It can be distinguished from the most similar congener, A. magna Kahl, 1932, by the presence of posterior protrusions, more membranelles in the posterior part of adoral zone (AZM2), and different SSU rRNA gene sequences. A. shini sp. n. is smaller in body size, only 35–40 × 25–30 μm in vivo, with four prominent ridges on the arched dorsal side, very similar with the “well-known” species A. steini Buddenbrock, 1920, in morphological characteristics, but it can be distinguished by the arrangement of frontoventral cirri and 121 nucleotide difference in the SSU rRNA gene sequences. Phylogenetic analyses based on the SSU rRNA gene sequences revealed the systematic positions of two new taxa and supported the validity of them as distinct species
Conjugation in Euplotes raikovi (Protista, Ciliophora): New Insights into Nuclear Events and Macronuclear Development from Micronucleate and Amicronucleate Cells
No full textCiliates form a distinct group of single‐celled eukaryotes that host two types of nuclei (micro and macronucleus) in the same cytoplasm and have a special sexual process known as conjugation, which involves mitosis, meiosis, fertilization, nuclear differentiation, and development. Due to their high species diversity, ciliates have evolved different patterns of nuclear events during conjugation. In the present study, we investigate these events in detail in the marine species Euplotes raikovi. Our results indicate that: (i) conjugation lasts for about 50 hours, the longest stage being the development of the new macronucleus (ca. 36 hours); (ii) there are three prezygotic micronuclear divisions (mitosis and meiosis I and II) and two postzygotic synkaryon divisions; and (iii) a fragment of the parental macronucleus fuses with the new developing macronucleus. In addition, we describe for the first time conjugation in amicronucleate E. raikovi cells. When two amicronucleate cells mate, they separate after about 4 hours without evident nuclear changes; when one amicronucleate cell mates with a micronucleate cell, the micronucleus undergoes regular prezygotic divisions to form migratory and stationary pronuclei, but the two pronuclei fuse in the same cell. In the amicronucleate cell, the parental macronucleus breaks into fragments, which are then recovered to form a new functional macronucleus. These results add new information on the process of conjugation in both micronucleate and amicronucleate Euplotes cells
Characterization of the macronuclear and micronuclear pheromone genes of Euplotes raikovi reveals the origin of the mating type genetic diversity
No full textCiliates produce diffusible, cell-type-specific pheromones to regulate growth and mating. In Euplotes, these signaling molecules belong to species-specific families of disulfide-rich and structurally homologous proteins. Pheromones are co-dominantly expressed by genes in the somatic macronucleus (MAC), whereas their allelic diversity originates from the mating type locus in the germline micronucleus (MIC). During MAC development in sexual process, the MIC-derived diversity of specific alleles is rearranged via macronucleus-destined sequences (MDSs) assembly. While many MAC pheromones are well characterized, their MIC precursors and rearrangement process remain unknown. Here, we identified two MAC pheromone genes (mac-er-13/14) of E. raikovi, and two MIC regions (19 kb in total) containing 10 MDSs that assemble into mac-er-13. These MDSs are separated by internal eliminated sequences (234-3345 bp). The shortest MDSs (9-36 bp) encode the secreted region of pheromone, while longer MDSs (44-419 bp) encode other regions. Considering that the secreted regions show a higher sequence variation and the shorter MDSs have higher probability of alternative processing or imprecise assembly, we hypothesize that the high sequence variability of the macronuclear pheromone genes, which underlies the large number of mating types in E. raikovi, may result from alternative processing or imprecise assembly of these short MDSs
Timing and characteristics of nuclear events during conjugation and genomic exclusion in Paramecium multimicronucleatum
No full textCiliated protists are ideal material for studying the origin and evolution of sex, because of their nuclear dimorphism (containing both germline micronucleus and somatic macronucleus in the same cytoplasm), special sexual processes (conjugation and autogamy), and high diversity of mating-type systems. However, the study of sexual process is limited to only a few species, due to the difficulties in inducing or observing conjugation. In the present study, we investigate the conjugation process in Paramecium multimicronucleatum: (1) of the three prezygotic divisions, all micronuclei undergo the first two divisions (meiosis I, II), while a variable number of nuclei undergo the third division (mitosis); (2) the synkaryon divides three times after fertilization, giving rise to eight products that differentiate into four macronuclear anlagen and four micronuclei; (3) cells restore the vegetative stage after two successive cell fissions during which the macronuclear anlagen are distributed into daughter cells without division, while micronuclei divide mitotically; (4) the parental macronucleus begins to fragment following the first meiotic division and finally degenerates completely; (5) the entire process takes about 110 h, of which about 85 h are required for macronuclear development. In addition, we describe for the first time the process of genomic exclusion occurring between amicronucleate and micronucleate cells of P. multimicronucleatum, during which the micronucleate cell contributes a pronucleus to the amicronucleate cell, resulting in both exconjugants being homozygotes. These results provide new insights into the diversity of sexual processes and lay an important cytological basis for future in-depth studies of mating systems in ciliates
Primary Structure and Coding Genes of Two Pheromones from the Antarctic Psychrophilic Ciliate, Euplotes focardii
No full textIn ciliates, diffusible cell type-specific pheromones regulate cell growth and mating phenomena acting competitively in both autocrine and heterologous fashion. In Euplotes species, these signaling molecules are represented by species-specific families of structurally homologous small, disulfide-rich proteins, each specified by one of a series of multiple alleles that are inherited without relationships of dominance at the mat-genetic locus of the germinal micronuclear genome, and expressed as individual gene-sized molecules in the somatic macronuclear genome. Here we report the 85-amino acid sequences and the full-length macronuclear nucleotide coding sequences of two pheromones, designated Ef-1 and Ef-2, isolated from the supernatant of a wild-type strain of a psychrophilic species of Euplotes, E. focardii, endemic to Antarctic coastal waters. An overall comparison of the determined E. focardii pheromone and pheromone-gene structures with their homologs from congeneric species provides an initial picture of how an evolutionary increase in the complexity of these structures accompanies Euplotes speciation
Fluorescent sensing of anions based on excited state intramolecular proton transfer in N-(3-hydroxy-2-naphthamido)-N'-phenylthiourea
No full textA neutral N-amidothiourea-based excited state intramolecular proton transfer (ESIPT) anion receptor bearing an o-hydroxynaphthamide fluorophore and a thiourea binding site, N-(3-hydroxy-2-naphthamide)-N'-phenylthiourea (1a), was designed and synthesized. Fluorescence and absorption response of 1a toward anions were assessed in acetonitrile. IR and NMR experiments indicated that the "OHa <-O=C" intramolecular hydrogen bond (IHB) in 1a was weak so that it only exhibited the short-wavelength normal emission other than ESIPT fluorescence. Due to the high anion binding affinity of the N-amidothiourea binding site and the formation of a hydrogen binding network in the 1a-anion complex, 1a underwent structural change upon anion binding that strengthens the "OHa <-O=C" IHB, leading to the ESIPT and the observation of the long-wavelength ESIPT emission whereas the normal fluorescence is quenched. On the basis of NMR and fluorescence titrations and control experiments with model compounds, a sensing mechanism of the anion-binding-induced ESIPT was proposed
Time-course analysis of nuclear events during conjugation in the marine ciliate Euplotes vannus and comparison with other ciliates (Protozoa, Ciliophora)
No full textThe widely reported but poorly studied ciliate family Folliculinidae (Protozoa, Ciliophora, Heterotrichea): a revision with notes on its taxonomy, morphology and phylogenetic relationships
No full textCiliates of the heterotrich family Folliculinidae are widespread in various habitats and are distinguished by their transparent loricae of various shapes, conspicuous peristomial lobes, and dimorphic life cycles. They usually attach firmly to the surface of substrates, feed on bacteria and microalgae, and play a significant role in energy flow and material cycling in the microbial food web. However, little is known regarding their biodiversity and systematics. In this work, we establish the terminology of the family Folliculinidae and select six crucial features for genus recognition. Based on previous studies, we revise the classification of Folliculinidae, supply improved diagnoses for each of the 33 folliculinid genera, and provide a key to their identification. Moreover, phylogenetic analyses based on small subunit ribosomal DNA (SSU rDNA) sequences revealed that the family is monophyletic and comprises two subclades (subclades I II) which can be identified by the flexibility of their peristomial lobes and the sculpturing of their necks. Furthermore, we investigate the evolutionary relationships of folliculinids using the six chosen generic features
