海鞘类天然产物的最新研究进展

近年来,从海洋生物海鞘中分离出很多结构新颖而又有显著生理活性的天然化合物,引起了有机化学家和药学家的关注。本文综述了国内外最近几年对海鞘类天然产物的研究进展,并按化学结构类型进行了分类。     

Involvement of vasa homolog in germline recruitment from coelomic stem cells in budding tunicates

We investigated the mechanism by which germline cells are recruited in every asexual reproductive cycle of the budding tunicate Polyandrocarpa misakiensis using a vasa homolog (PmVas) as the germline-specific probe. A presumptive gonad of Polyandrocarpa arose as a loose cell aggregate in the ventral hemocoel of a 1-week-old developing zooid. It developed into a compact clump of cells and then separated into two lobes, each differentiating into the ovary and the testis. The ovarian tube that was formed at the bottom of the ovary embedded the oogonia and juvenile oocytes, forming the germinal epithelium. PmVas was expressed strongly by loose cell aggregates, compact clumps, and peripheral germ cells in the testis and germinal epithelium. No signals were detected in growing buds and less than 1-week-old zooids, indicating that germ cells arise de novo in developing zooids of P. misakiensis. Cells of the loose cell aggregates were 5–6 μm in diameter. They looked like undifferentiated hemoblasts in the hemocoel. To examine the involvement of PmVas in the germline recruitment at postembryonic stages, both growing buds and 1-week-old developing zooids were soaked with double-stranded PmVas RNA. The growing buds developed into fertile zooids expressing PmVas, whereas the 1-week-old zooids developed into sterile zooids that did not express PmVas. In controls (1-week-old zooids) soaked with double-stranded lacZ RNA, the gonad developed normally. These results strongly suggest that in P. misakiensis, PmVas plays a decisive role in switching from coelomic stem cells to germ cells.     

Muscle development in Ciona intestinalis requires the b-HLH myogenic regulatory factor gene Ci-MRF

The activity of myogenic regulatory factor (MRF) genes is essential for vertebrate muscle development, whereas invertebrate muscle development is largely independent of MRF function. This difference indicates that myogenesis is controlled by distinct regulatory mechanisms in these two groups of animals. Here we used overexpression and gene knockdown to investigate the role in embryonic myogenesis of the single MRF gene of the invertebrate chordate Ciona intestinalis (Ci-MRF). Injection of Ci-MRF mRNA into eggs resulted in increased embryonic muscle-specific gene activity and revealed the myogenic activity of Ci-MRF by inducing the expression of four muscle marker genes, Acetylcholinesterase, Actin, Troponin I, and Myosin Light Chain in non-muscle lineages. Conversely, inhibiting Ci-MRF activity with antisense morpholinos down-regulated the expression of these genes. Consistent with the effects of morpholinos on muscle gene activity, larvae resulting from morpholino injection were paralyzed and their "muscle" cells lacked myofibrils. We conclude that Ci-MRF is required for larval tail muscle development and thus that an MRF-dependent myogenic regulatory network probably existed in the ancestor of tunicates and vertebrates. This possibility raises the question of whether the earliest myogenic regulatory networks were MRF-dependent or MRF-independent.     

Telomerase maintained in self-renewing tissues during serial regeneration of the urochordate Botryllus schlosseri

Telomerase is critical for the protection of germ line and stem cell chromosomes from fatal shortening during replication. In most organisms, telomerase activity is suppressed in progressively committed cells and falls to basal rates in terminally differentiated lineages. The colonial ascidian Botryllus schlosseri propagates asexually and sexually, presumably from pools of stem cells that self-renew throughout the 2- to 5-year colony life span. Asexual budding takes place continuously from the parental body wall. When the colony reaches a critical size, sexual reproduction commences with the generation of gonads. Here, we establish the existence of 6-15 kb telomeres on the ends of Botryllus chromosomes. We develop a real-time quantitative PCR telomeric repeat amplification protocol (TRAP) assay that reliably detects 0.2-100 TPG units in cells and tissues. We find highest levels of enzymatic activity in the gonads, developing embryos, and tissues containing the earliest asexual buds. Telomerase activity appears to be suppressed in later buds during organogenesis and falls to basal rates in mature zooids. We postulate that this pattern reflects maximum telomere restoration in somatic stem cells of early buds and suppression of telomerase activity in progenitors and terminally differentiated cells, indicative of an alternate role for stem cells as repeated body regenerators in colonial life histories.     

Jak-STAT regulation of male germline stem cell establishment during Drosophila embryogenesis

Germline stem cells (GSCs) in Drosophila are descendants of primordial germ cells (PGCs) specified during embryogenesis. The precise timing of GSC establishment in the testis has not been determined, nor is it known whether mechanisms that control GSC maintenance in the adult are involved in GSC establishment. Here, we determine that PGCs in the developing male gonad first become GSCs at the embryo to larval transition. This coincides with formation of the embryonic hub; the critical signaling center that regulates adult GSC behavior within the stem cell microenvironment (niche). We find that the Jak-STAT signaling pathway is activated in a subset of PGCs that associate with the newly-formed embryonic hub. These PGCs express GSC markers and function like GSCs, while PGCs that do not associate with the hub begin to differentiate. In the absence of Jak-STAT activation, PGCs adjacent to the hub fail to exhibit the characteristics of GSCs, while ectopic activation of the Jak-STAT pathway prevents differentiation. These findings show that stem cell formation is closely linked to development of the stem cell niche, and suggest that Jak-STAT signaling is required for initial establishment of the GSC population in developing testes.     

Retinal is the essential form of retinoid for storage and transport in the adult of the ascidian Halocynthia roretzi

Retinoids in the organs (gonad [GND], body wall muscle [BWM], hepatopancreas [HP], gill, hemolymph cells and hemolymph plasma) of the adult ascidian Halocynthia roretzi were analyzed by high performance liquid chromatography. Retinal (RAL) occurred in every organ examined, and most of RAL (≥99%) was localized in the GND and BWM. None of the organs contained significant amounts of retinol (ROL) or retinyl ester (RE). Lipid droplets, which are characteristic of stellate cells (RE-storing cells of vertebrates), could not be found in the GND, BWM and HP by microscopic observations. These results indicate that this ascidian lacks the RE-storing mechanism, which is ubiquitous in adult vertebrates. The amount and localization of RAL showed the annual change in relation to the reproductive cycle. During summer, the growing season, RAL was present in both GND and BWM at a ratio of about 3:2. From summer to winter, RAL in the GND gradually increased, concomitant with the decrease of RAL in the BWM. In winter, the spawning season, most of RAL was present in the GND (ca. 98%). RAL appears to be accumulated first in the BWM and transported to oocytes accompanying yolk accumulation. ROL and RE were not implicated in the storage and transport of retinoids. The results in the present research strongly suggest that retinoic acid (RA) is produced by the two-step enzymatic reaction: carotenoid cleavage to RAL followed by RAL oxidation to RA and that the prevertebrate chordate lacks ROL-metabolizing systems.     

Cadiolides J–M,antibacterial polyphenyl butenolides from the Korean tunicate Pseudodistoma antinboja

Activity-guided fractionations of the tunicate Pseudodistoma antinboja yielded four new compounds of the cadiolide class (cadiolides J–M, 1, 3–5) along with a known one (cadiolide H, 2). The structures were defined by spectroscopic methods including X-ray crystallographic analysis. These compounds were evaluated for their antibacterial activity and exhibited potent antibacterial activity against all of the drug resistant strains tested with MICs comparable to those of marketed drugs such as vancomycin and linezolid.     

The Oikopleura coenocyst, a unique chordate germ cell permitting rapid, extensive modulation of oocyte production

The ability to adjust reproductive output to environmental conditions is important to the fitness of a species. The semelparous, chordate, Oikopleura dioica, is particularly adept in producing a highly variable number of oocytes in its short life cycle. Here we show that this entails an original reproductive strategy in which the entire female germline is contained in a single multinucleate cell, the "coenocyst". After an initial phase of syncytial nuclear proliferation half of the nuclei entered meiosis whereas the other half became highly polyploid. The inner F-actin network, with associated plasma membranes, formed a highly ramified infrastructure in which each meiotic nucleus was contained in a pseudo-compartmentalized pro-oocyte linked to the common cytoplasm via ring canals. At a set developmental time, a subset of the pro-oocytes was selected for synchronous growth and the common coenocyst cytoplasm was equally partitioned by transfer through the ring canals. Examination of related species indicated that the coenocyst arrangement is a conserved feature of Appendicularian oogenesis allowing efficient numerical adjustment of oocyte production. As Appendicularia are the second most abundant class of zooplankton, with a world-wide distribution, the coenocyst is clearly a common and successful reproductive strategy on a global scale.     

The colonial ascidian Didemnum sp. A: Current distribution, basic biology and potential threat to marine communities of the northeast and west coasts of North America

Didemnum sp. A is a colonial ascidian with rapidly expanding populations on the east and west coasts of North America. The origin of Didemum sp. A is unknown. Populations were first observed on the northeast coast of the U.S. in the late 1980s and on the west coast during the 1990s. It is currently undergoing a massive population explosion and is now a dominant member of many subtidal communities on both coasts. To determine Didemnum sp. A's current distribution, we conducted surveys from Maine to Virginia on the east coast and from British Columbia to southern California on the west coast of the U.S. between 1998 and 2005. In nearshore locations Didemnum sp. A currently ranges from Eastport, Maine to Shinnecock Bay, New York on the east coast. On the west coast it has been recorded from Humboldt Bay to Port San Luis in California, several sites in Puget Sound, Washington, including a heavily fouled mussel culture facility, and several sites in southwestern British Columbia on and adjacent to oyster and mussel farms. The species also occurs at deeper subtidal sites (up to 81 m) off New England, including Georges, Stellwagen and Tillies Banks. On Georges Bank numerous sites within a 230 km² area are 50-90% covered by Didemnum sp. A; large colonies cement the pebble gravel into nearly solid mats that may smother infaunal organisms. These observations suggest that Didemnum sp. A has the potential to alter marine communities and affect economically important activities such as fishing and aquaculture.     

Action of the Tunicate locus on maize floral development

The co-dominant Tunicate (Tu) mutation in maize causes nonreproductive structures in both the male and female inflorescences to be enlarged. This mutation also affects sex determination, permitting the development of pistils in the normally staminate tassel. In order to characterize the role of the normal tu gene product, we have analysed genetic interaction between Tu and other mutations that perturb specific stages of floral development. Synergistic interactions observed suggested that the tu product functions in at least three stages of floral development–determination of spikelet primordia, differentiation of non-reproductive organs and pistil abortion in the tassel. © 1994 Wiley-Liss, Inc.     

海鞘与蓝藻的共生关系

互利共生是指两种不同生物之间所形成的紧密互利关系。文章以海鞘与蓝藻共生关系的典型实例为代表,介绍了蓝藻与海鞘共生关系的发现过程;蓝藻在海鞘间世代传递的机制;两者在共生关系中可能存在的生理功能,如蓝藻共生体通过碳固定、参与氮循环和产生代谢物等方式为海鞘提供营养并参与海鞘的防御活动,海鞘可供给共生体生长所需的一部分氮素营养和提供抗紫外线的保护等。对海鞘与蓝藻共生关系的研究将有助于理解生物进化过程,以及为进一步开发利用海洋生物之间的共生关系提供研究基础。     

Ultrastructure of sensory receptors in ascidian tadpoles

Summary An analysis of fine structure and function was conducted on three types of receptors in the cerebral vesicle of two species of ascidian tadpoles (Ciona intestinalis andDistaplia occidentalis). Theocellus is composed of one pigmented, cup-shaped supportive cell, an estimated 15–20 sensory cells, and three lens cells, each with a large body of granules (glycogen ?). The outer segments of the photoreceptoral processes are modified cilia, one per sensory cell, consisting of many lamellae, formed by infoldings of the ciliary membranes, and axonemes of 9 + 0 doublets of microtubules. The lamellae are homologous to retinal disks of vertebrates.Hydrostatic pressure (?)receptors are modified cilia containing tubules which open to the lumen of the cerebral vesicle. These receptors closely resemble the globular, ciliary processes of coronet cells in the saccus vasculosus of fishes. Thestatocyte is a onecell gravity receptor. The part extending into the lumen of the brain contains the nucleus and a large black body which is thought to function as a float. The foot-piece of the cell is firmly anchored in the brain wall; the neck is probably the site of generation of signals. This study was supported by grant 10292 from the USPHS. The senior author held an appointment in the Miller Institute for Basic Research in Science, University of California, during part of the investigation. We appreciate the assistance of Emily Reid, artist; Jean L. Brandenburger, research associate; Professor Gerald Westheimer, consultant; Charles Spowart, collector; and the UCLA Brain Information Service, and the hospitality of Dr. Robert L. Fernald, Director, Friday Harbor Laboratories, University of Washington, U.S.A.     

Tunicate tails, stolons, and the origin of the vertebrate trunk

Tunicates are primitive chordates that develop a transient 'tail' in the larval stage that is generally interpreted as a rudimentary version of the vertebrate trunk. Not all tunicates have tails, however. The groups that lack them, salps and pyrosomes, instead have a trunk-like reproductive stolon located approximately where the tail would otherwise be. In salps, files of blastozooids are formed along the sides of the stolon. The tail and caudal trunk in more advanced chordates could have evolved from a stolon of this type, an idea referred to here as the 'stolon hypothesis'. This means the vertebrate body could be a composite structure, since there is the potential for each somite to incorporate elements originally derived from a complete functional zooid. If indeed this has occurred, it should be reflected in some fashion in gene expression patterns in the vertebrate trunk. Selected morphological and molecular data are reviewed to show that they provide some circumstantial support for the stolon hypothesis. The case would be stronger if it could be demonstrated that salps and/or pyrosomes are ancestral to other tunicates. The molecular phylogenies so far available generally support the idea of a pelagic ancestor, but offer only limited guidance as to which of the surviving pelagic groups most closely resembles it. The principal testable prediction of the stolon hypothesis is that head structures (or their homologues) should be duplicated in series in the trunk in advanced chordates, and vice versa, i.e. trunk structures should occur in the head. The distribution of both rhabdomeric photoreceptors and nephridia in amphioxus conform with this prediction. Equally striking is the involvement of the Pax2 gene in the development of both the inner ear and nephric ducts in vertebrates. The stolon hypothesis would explain this as a consequence of the common origin of otic capsules and excretory ducts from atrial rudiments: from the paired rudiments of the parent oozooid in the case of the otic capsule (these express Pax2 according to recent ascidian data), and from tubular rudiments in the stolon in the case of the excretory ducts.     

Characterization of vanadium-binding sites of the vanadium-binding protein Vanabin2 by site-directed mutagenesis

Background

Vanabins are a unique protein family of vanadium-binding proteins with nine disulfide bonds. Possible binding sites for VO²⁺ in Vanabin2 from a vanadium-rich ascidian Ascidia sydneiensis samea have been detected by nuclear magnetic resonance study, but the metal selectivity and metal-binding ability of each site was not examined.

Methods

In order to reveal functional contribution of each binding site, we prepared several mutants of Vanabin2 by in vitro site-directed mutagenesis and analyzed their metal selectivity and affinity by immobilized metal-ion affinity chromatography and Hummel Dreyer method.

Results

Mutation at K10/R60 (site 1) markedly reduced the affinity for VO²⁺. Mutation at K24/K38/R41/R42 (site 2) decreased the maximum binding number, but only slightly increased the overall affinity for VO²⁺. Secondary structure of both mutants was the same as that of the wild type as assessed by circular dichroism spectroscopy. Mutation in disulfide bonds near the site 1 did not affect its high affinity binding capacity, while those near the site 2 decreased the overall affinity for VO²⁺.

General significance

These results suggested that the site 1 is a high affinity binding site for VO²⁺, while the site 2 composes a moderate affinity site for multiple VO²⁺.