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PubMed1985

Synthesis and possible role of proteoglycans during Volvox development.

During the embryonic development of Volvox the cells synthesize proteoglycans of relatively small molecular size up to 14 h after inversion. In the following stages there is a gradual transition towards larger forms, due to an increase in sulfated sugar moieties. The presence of sulfate in the medium was obligatory for this transition. None of these molecules could be dissociated in 4.0 M GuHCl, neither could they be digested by glycosidases. Of all proteases examined subtilisin, a bacterial protease, digested the proteoglycans. The possible role of proteoglycans during Volvox morphogenesis has been examined. Somatic proteoglycans caused spheroids to start their swimming earlier, although they inhibited their expansion. Spheroids cultured in sulfate-free medium start their swimming 24 h later than controls, but the addition of somatic proteoglycans to the sulfate-free medium could induce motility almost as well as in the normal forms. Embryonic weight proteoglycans caused aggregation of the spheroids, an effect that also appeared in the sulfate-free medium.

Animals↗

PubMed1979

Morphogenesis in Volvox: analysis of critical variables.

Inversion, the process by which Volvox embryos turn inside out, was analyzed by a combination of geometrical and experimental techniques. It was shown that simple geometric figures are adequate to represent cell shapes during inversion and that cell volumes remain constant as cell shapes change and the embryo inverts. The first stage of inversion, phialopore opening, results from the release of compressive forces as the embryo withdraws from its surrounding vesicle during a two-stage contraction of each cell around its radial axis. Premature phialopore opening occurs when withdrawal of the embryo from the vesicle is elicited artificially by exposure to either calcium ionophore or hypertonic solutions. The major event of inversion, generation of negative curvature, requires both microtubule-driven elongation of cells (to produce a classical "flask" shape) and cytochalasin-sensitive active migration of cytoplasmic bridges to the outermost ends of flask cells. Colchicine, cyclic GMP and isobutyl methyl xanthine (individually) block both normal elongation and bridge migration; cytochalasin D blocks bridge migration selectively. Flask cell formation and bridge migration are adequate to account for the negative curvature observed. An asymmetric bending of flask cell stalks along the ring of maximum curvature accounts for the fact that the embryo is not constricted in a "purse-string" fashion as negative curvature is generated. Inversion of the posterior hemisphere involves an elastic snap-through resulting from a combination of compressive stresses generated by inversion of the anterior hemisphere and the circumferential restraint imposed by cells at the equator. We conclude that the observed changes in cell shape and the migration of cytoplasmic bridges are the result of an ordered process of membrane-cytoskeletal interactions, and both necessary and sufficient to account for the morphogenetic process of inversion in Volvox.

Animals↗

PubMed1999

Two light-activated conductances in the eye of the green alga Volvox carteri.

Photoreceptor currents of the multicellular green alga Volvox carteri were analyzed using a dissolver mutant. The photocurrents are restricted to the eyespot region of somatic cells. Photocurrents are detectable from intact cells and excised eyes. The rhodopsin action spectrum suggests that the currents are induced by Volvox rhodopsin. Flash-induced photocurrents are a composition of a fast Ca2+-carried current (PF) and a slower current (PS), which is carried by H+. PF is a high-intensity response that appears with a delay of less than 50 micros after flash. The stimulus-response curve of its initial rise is fit by a single exponential and parallels the rhodopsin bleaching. These two observations suggest that the responsible channel is closely connected to the rhodopsin, both forming a tight complex. At low flash energies PS is dominating. The current delay increases up to 10 ms, and the PS amplitude saturates when only a few percent of the rhodopsin is bleached. The data are in favor of a second signaling system, which includes a signal transducer mediating between rhodopsin and the channel. We present a model of how different modes of signal transduction are accomplished in this alga under different light conditions.

Biophysical Phenomena↗

PubMed2002

Palindromic repetitive elements in the mitochondrial genome of Volvox.

Group I introns were found in the cob and cox I genes of Volvox carteri. These introns contain tandem arrays of short palindromic sequences that are related to each other. Inspection of other regions in the mtDNA revealed that similar palindromic repetitive sequences are dispersed in the non-protein coding regions of the mitochondrial genome. Analysis of the group I intron in the cob gene of another member of Volvocaceae, Volvox aureus, has shown that its sequence is highly homologous to its counterpart in V. carteri with the exception of a cluster of palindromic sequences not found in V. carteri. This indicates that the palindromic clusters were inserted into the introns after divergence of the two species, presumably due to frequent insertions of the palindromic elements during evolution of the Volvocaceae. Possible involvement of the palindromic repetitive elements in the molecular evolution of functional RNAs is discussed.

Base Sequence↗

PubMed1973

Genetic control of development in Volvox: isolation and characterization of morphogenetic mutants.

Morphogenetic mutants of the colonial green alga, Volvox carteri f. nagariensis, were induced by chemical mutagenesis. The 68 independent mutants are classified into 12 readily identifiable phenotypes affecting various stages of asexual development. Nine of the mutants are temperature sensitive with normal development at 25 degrees and mutant development occurring at 35 degrees . Some mutant genes appear to be involved in the regulation of differentiation or the stability of the differentiated state. Other mutations occur in genes apparently responsible for structural components of dividing cells or adult colonies. Two mutants affect different aspects of the posterior-anterior polarization of the mature colony. One mutation affects a gene which acts very early in colonial development, but affects the appearance of the mature colony. The mutants isolated demonstrate the feasibility of using Volvox to study the genetic control of early steps in embryogenesis.

Alkanesulfonates↗

PubMed1981

Sulfation of a cell surface glycoprotein correlates with the developmental program during embryogenesis of Volvox carteri.

A sulfated cell surface glycoprotein with an apparent molecular weight of 185,000 is synthesized in the multicellular organism Volvox only during the limited period of embryogenesis. The lifetime of sulfate residues on this glycoprotein is very short (half-life about 20 min). Production of this sulfated glycoprotein sharply decreases to a minimum shortly before the onset of the differentiating cell cleavage--e.g., in asexual development, before the 32-cell embryo divides. It is demonstrated that the sulfated glycoprotein behaves in many respects as would the hypothetical cell surface component postulated by a recently published model [Sumper, M. (1979) FEBS Lett. 107, 241-246], which proposes an explanation for the cell-counting mechanism and the spatial control of differentiation that is operative in Volvox embryogenesis.

Cell Adhesion↗

PubMed1999

Response to the sexual pheromone and wounding in the green alga volvox: induction of an extracellular glycoprotein consisting almost exclusively of hydroxyproline.

The extracellular matrix (ECM) of Volvox is modified during development or in response to external stimuli, like the sex-inducing pheromone. It has recently been demonstrated that a number of genes triggered by the sex-inducing pheromone are also inducible by wounding. By differential screening of a cDNA library, a novel gene was identified that is transcribed in response to the pheromone. Its gene product was characterized as an ECM glycoprotein with a striking feature: it exhibits a hydroxyproline content of 68% and therefore is an extreme member of the family of hydroxyproline-rich glycoproteins (HRGPs). HRGPs are known as constituents of higher plant ECMs and seem to function as structural barriers in defense responses. The Volvox HRGP is also found to be inducible by wounding. This indicates that the wound response scenarios of higher plants and multicellular green algae may be evolutionary related.

Algal Proteins↗

PubMed1987

Nucleated assembly of Chlamydomonas and Volvox cell walls.

The Chlamydomonas reinhardtii cell wall is made up of hydroxyproline-rich glycoproteins, arranged in five distinct layers. The W6 (crystalline) layer contains three major glycoproteins (GP1, GP2, GP3), selectively extractable with chaotropic agents, that self-assemble into crystals in vitro. A system to study W6 assembly in a quantitative fashion was developed that employs perchlorate-extracted Chlamydomonas cells as nucleating agents. Wall reconstitution by biotinylated W6 monomers was monitored by FITC-streptavidin fluorescence and quick-freeze/deep-etch electron microscopy. Optimal reconstitution was obtained at monomer concentrations (0.2-0.3 mg/ml) well below those required for nonnucleated assembly. Assembly occurred from multiple nucleation sites, and faithfully reflected the structure of the intact W6 layer. Specificity of nucleated assembly was demonstrated using two cell-wall mutants (cw-2 and cw-15); neither served as a substrate for assembly of wild-type monomers. In addition, W6 sublayers were assembled from purified components: GP2 and GP3 coassembled to form the inner (W6A) sublayer; this then served as a substrate for self-assembly of GP1 into the outer (W6B) sublayer. Finally, evolutionary relationships between C. reinhardtii and two additional members of the Volvocales (Chlamydomonas eugametos and Volvox carteri) were explored by performing interspecific reconstitutions. Hybrid walls were obtained between C. reinhardtii and Volvox but not with C. eugametos, confirming taxonomic assignments based on structural criteria.

Cell Nucleus↗

PubMed1977

Cell shape changes and the mechanism of inversion in Volvox.

Inversion is a dominant aspect of morphogenesis in Volvox. In this process, the hollow, spheroidal Volvox embryo turns inside-out through a small opening called the phialopore to bring flagella from its inner to its outer surface. Analyses of intact, sectioned, and fragmented embryos by light, scanning electron, and transmission electron microscopy, suggest that shape changes preprogrammed into the cells cause inversion. First, cells throughout the embryo change from pear to spindle shape, which causes the embryo to contract and the phialopore to open. Then cells adjacent to the phialopore become flask-shaped, with long, thin stalks at their outer ends. Simultaneously, the cytoplasmic bridges joining all adjacent cells migrate from the midpoint of the cells to the stalk tips. Together, these changes cause the lips of cells at the phialopore margin to curl outward. Now cells progressively more distal to the phialopore become flask-shaped while the more proximal cells become columnar, causing the lips to curl progressively further over the surface of the embryo until the latter has turned completely inside-out. Fine structural analysis reveals a peripheral cytoskeleton of microtubules that is apparently involved in cellular elongation. Cell clusters isolated before inversion undergo a similar program of shape changes; this suggests that the changes in cellular shape are the cause rather than an effect of the inversion process.

Cell Division↗

PubMed1997

Differential targeting of closely related ECM glycoproteins: the pherophorin family from Volvox.

The alga Volvox carteri represents one of the simplest multicellular organisms. Its extracellular matrix (ECM) is modified under developmental control, e.g. under the influence of the sex-inducing pheromone that triggers development of males and females at a concentration below 10(-16) M. A novel ECM glycoprotein (pherophorin-S) synthesized in response to this pheromone was identified and characterized. Although being a typical member of the pherophorins, which are identified by a C-terminal domain with sequence homology to the sex-inducing pheromone, pherophorin-S exhibits a completely novel set of properties. In contrast to the other members of the family, which are found as part of the insoluble ECM structures of the cellular zone, pherophorin-S is targeted to the cell-free interior of the spherical organism and remains in a soluble state. A main structural difference is the presence of a polyhydroxyproline spacer in pherophorin-S that is linked to a saccharide containing a phosphodiester bridge between two arabinose residues. Sequence comparisons indicate that the self-assembling proteins that create the main parts of the complex Volvox ECM have evolved from a common ancestral gene.

Algal Proteins↗

PubMed2001

A link between DNA methylation and epigenetic silencing in transgenic Volvox carteri.

Epigenetic silencing of foreign genes introduced into plants poses an unsolved problem for transgenic technology. Here we have used the simple multicellular green alga VOLVOX: carteri as a model to analyse the relation of DNA methylation to transgenic silencing. VOLVOX: DNA contains on average 1.1% 5-methylcytosine and 0.3% N6-methyladenine, as revealed by electrospray mass spectrometry and phosphoimaging of chromatographically separated (32)P-labelled nucleotides. In two nuclear transformants of V.carteri, produced in 1993 by biolistic bombardment with a foreign arylsulphatase gene (C-ars), the transgene is still expressed in one (Hill 181), but not in the other (Hill 183), after an estimated 500-1000 generations. Each transformant clone contains multiple intact copies of C-ars, most of them integrated into the genome as tandem repeats. When the bisulphite genomic sequencing protocol was applied to examine two select regions of transgenic C-ars, we found that the inactivated copies (Hill 183) exhibited a high-level methylation (40%) of CpG dinucleotides, whereas the active copies (Hill 181) displayed low-level (7%) CpG methylation. These are average values from 40 PCR clones sequenced from each DNA strand in the two portions of C-ars. The observed correlation of CpG methylation and transgene inactivation in a green alga will be discussed in the light of transcriptional silencing.

5-Methylcytosine↗

PubMed1980

Lipid Composition and Metabolism of Volvox carteri.

The membrane structural lipids of somatic cells and gonidia isolated from Volvox carteri f. nagariensis spheroids have been characterized. The principal polar lipid components of both cell types are sulfoquinovosyl diglyceride, mono- and digalactosyl diglyceride, phosphatidylglycerol, phosphatidylethanolamine, and 1(3), 2-diacylglyceryl-(3)-O-4'-(N,N,N,-trimethyl)homoserine. Light-synchronized cultures of spheroids were shown to incorporate [(14)C]bicarbonate, [(35)S]sulfate, [(14)C]palmitic acid, and [(14)C]lauric acid into complex lipids. [(14)C]Palmitic acid was incorporated mainly into diacylglyceryltrimethylhomoserine and was not significantly modified by elongation or desaturation. In contrast, [(14)C]lauric acid was incorporated into a wider variety of complex lipids and was also converted into longer chain saturated and unsaturated fatty acids. Volvox is a promising system for studying the role of membranes in algal cellular differentiation.

Journal Article↗

PubMed2002

Evidence for autocatalytic cross-linking of hydroxyproline-rich glycoproteins during extracellular matrix assembly in Volvox.

The alga Volvox carteri is one of the simplest multicellular organisms, yet it has a surprisingly complex extracellular matrix (ECM), making Volvox suitable as a model system in which to study ECM self-assembly. Here, we analyze the primary structures and post-translational modifications of two main ECM components synthesized in response to sexual induction as well as wounding. These proteins are members of the pherophorin family with as yet unknown properties. They contain polyhydroxyproline spacers as long as 500 and 2750 residues. Even the highly purified proteins retain the capacity to self-assemble and cross-link, producing an insoluble fibrous network in an apparently autocatalytic reaction. This pherophorin-based network is located within the deep zone of the ECM. A molecular genetic search for additional members of the pherophorin family indicates that at least nine different pherophorin species can be expected to serve as precursors for ECM substructures. Therefore, the highly diversified members of the pherophorin family represent region-specific morphological building blocks for ECM assembly and cross-linking.

Adaptation, Physiological↗

PubMed1986

Heat shock elicits production of sexual inducer in Volvox.

In the green alga Volvox carteri, heat shock had an unusual and adaptive effect mediated by induced production of a well-defined effector molecule. Females of this species normally reproduce asexually in the absence of a potent sexual inducer produced by mature sexual males, but they generated egg-bearing sexual daughters after a brief exposure to elevated temperatures. This response involved an "autoinduction" of sexuality, in which heat-shocked somatic cells made and released the sexual inducer, which then redirected development of the reproductive cells. Males, including a sterile mutant incapable of producing inducer in the usual manner, also produced the inducer in response to heat shock. The phenomenon probably is of significance in the wild, where Volvox reproduces asexually in temporary ponds in spring but becomes sexual and produces dormant, overwintering zygotes before the ponds dry up in the summer heat.

Adaptation, Physiological↗

PubMed1999

regA, a Volvox gene that plays a central role in germ-soma differentiation, encodes a novel regulatory protein.

Volvox has two cell types: mortal somatic cells and immortal germ cells. Here we describe the transposon-tagging, cloning and characterization of regA, which plays a central role as a master regulatory gene in Volvox germ-soma differentiation by suppressing reproductive activities in somatic cells. The 12.5 kb regA transcription unit generates a 6,725 nucleotide mRNA that appears at the beginning of somatic cell differentiation, and that encodes a 111 kDa RegA protein that localizes to the nucleus, and has an unusual abundance of alanine, glutamine and proline. This is a compositional feature shared by functional domains of many 'active' repressors. These findings are consistent with the hypothesis that RegA acts in somatic cells to repress transcription of genes required for growth and reproduction, including 13 genes whose products are required for chloroplast biogenesis.

Algal Proteins↗

PubMed1986

The extracellular matrix of Volvox: a comparative study and proposed system of nomenclature.

The structure of the extracellular matrix (ECM) of representatives of all four sections of the genus Volvox was examined by a combination of light- and electron-microscopic methods. On the basis of these observations, plus published descriptions of aspects of ECM organization in other members of the order Volvocales, a system of nomenclature is proposed, to facilitate discussion of comparative morphology and phylogeny of the ECM in the order. In this system the ECM is divided into four main zones: the flagellar zone (FZ), which consists of attachments to and specializations of the ECM around the flagella; the boundary zone (BZ), which consists of portions of the ECM that (except in periflagellar regions) are continuous over the surface of the organism and are not structurally continuous with deeper layers; the cellular zone (CZ), which consists of specializations, other than those of the FZ, around individual cells; and the deep zone (DZ), which consists of components that fill the central region of the organism, internal to CZ. An empirically based set of hierarchical subdivisions of these zones is then proposed that permits specific identification of most morphologically distinct ECM components. The fact that not all zones and subzones are present in all members of the order means that this system permits identification of those ECM structures that have been gained or lost during Volvocalean evolution. Species-specific differences in the structure of virtually all aspects of the ECM were seen among the Volvox species examined in this study. However, the fact that such differences cannot always be used as diagnostic characters for the four divisions of the genus was demonstrated by the observation that in certain ECM features two members of the same division (V. carteri f. nagariensis and V. carteri f. weismannia) differ markedly in structure from one another, with one member of the pair resembling a member of another division. Thus many details of ECM organization appear to be under separate control, and capable of independent evolution.

Extracellular Matrix↗

PubMed1976

The DNA of Volvox carteri: a biophysical and biosynthetic characterization.

Various biophysical and biosynthetic characteristics of deoxyribonucleic acid (DNA) from Volvox carteri are examined. The DNA from three strains (HK-10,NB-7 and KA-1) is compared, and all strains are shown to contain at least two distinct DNA species which band at densities of 1.714-1.715 and 1.704-1.705 g/cm3 in neutral CsCl and correspond to nuclear and "cytoplasmic" DNA, respectively. Base compositions calculated from these densities, 55-56% G+C for nuclear DNA, and 45-46% G+C for cytoplasmic DNA, are in close agreement with % G+C values estimated from thermal denaturation data. DNA from strain KA-1 has a third component with a buoyant density of 1.693 g/cm3. DNA synthesis is analysed using radioactively labelled heterogeneously grown strains of Volvox carteri and profiles obtained following preparative CsCl density gradient centrifugation are presented. In addition, dissimilarities in patterns of DNA synthesis at various periods in the asexual life cycle are reported for synchronous cultures of strain HK-10. These differences in temporal patterns of DNA synthesis clearly indicate that while nuclear DNA is make to some degree throughout the life cycle, cytoplasmic DNA synthesis appears to occur only at discrete intervals.

Animals↗

PubMedDate not supplied

[Dormant stages of the green flagellate Volvox in a natural habitat].

Reproduction and formation of resistant dormant spores in Volvox aureus (Ehr.) and V. tertius (Meyer) were studied in a biocoenosis of a transient pool in July-August 1996 and 1997. Under these conditions, the populations of two Volvox species had species-specific reproductive features. In the V. tertius population, a relatively small amount of male individuals and dormant spores (zygospores) appeared sporadically. On the contrary, in the V. aureus population, dormant parthenospores were formed, but male individuals were never observed.

Chlorophyta↗