Oocyte activation and phospholipase C zeta (PLCζ): diagnostic and therapeutic implications for assisted reproductive technology

ABSTRACT: Infertility affects one in seven couples globally and has recently been classified as a disease by the World Health Organisation (WHO). While in-vitro fertilisation (IVF) offers effective treatment for many infertile couples, cases exhibiting severe male infertility (19-57%) often remain difficult, if not impossible to treat. In such cases, intracytoplasmic sperm injection (ICSI), a technique in which a single sperm is microinjected into the oocyte, is implemented. However, 1-5% of ICSI cycles still fail to fertilise, affecting over 1000 couples per year in the UK alone. Pregnancy and delivery rates for IVF and ICSI rarely exceed 30% and 23% respectively. It is therefore imperative that Assisted Reproductive Technology (ART) protocols are constantly modified by associated research programmes, in order to provide patients with the best chances of conception. Prior to fertilisation, mature oocytes are arrested in the metaphase stage of the second meiotic division (MII), which must be alleviated to allow the cell cycle, and subsequent embryogenesis, to proceed. Alleviation occurs through a series of concurrent events, collectively termed 'oocyte activation'. In mammals, oocytes are activated by a series of intracellular calcium (Ca2+) oscillations following gamete fusion. Recent evidence implicates a sperm-specific phospholipase C, PLCzeta (PLCζ), introduced into the oocyte following membrane fusion as the factor responsible. This review summarises our current understanding of oocyte activation failure in human males, and describes recent advances in our knowledge linking certain cases of male infertility with defects in PLCζ expression and activity. Systematic literature searches were performed using PubMed and the ISI-Web of Knowledge. Databases compiled by the United Nations and World Health Organisation databases (UNWHO), and the Human Fertilization and Embryology Authority (HFEA) were also scrutinised. It is clear that PLCζ plays a fundamental role in the activation of mammalian oocytes, and that genetic, molecular, or biochemical perturbation of this key enzyme is strongly linked to human infertility where oocyte activation is deficient. Consequently, there is significant scope for our understanding of PLCζ to be translated to the ART clinic, both as a novel therapeutic agent with which to rescue oocyte activation deficiency (OAD), or as a prognostic/diagnostic biomarker of oocyte activation ability in target sperm samples.     

The post-embryonic development of Remipedia (Crustacea)—additional results and new insights

The post-embryonic development of a species of the enigmatic crustacean group Remipedia is described in detail for the first time under various aspects. Applying a molecular approach, we can clearly prove the species identity of the larvae as belonging to Pleomothra apletocheles. We document the cellular level of several larval stages and the differentiation of segments, limbs, and the general body morphology applying the techniques of confocal laser scanning microscopy and scanning electron microscopy. In addition, we document the swimming behavior and the peculiar movements of the naupliar appendages. A comparison of our results with published data on other Crustacea and their larval development tentatively supports ideas about phylogenetic affinities of the Remipedia to the Malacostraca. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. An erratum to this article can be found at     

The generalist Inga subnuda subsp. luschnathiana (Fabaceae): negative effect of floral visitors on reproductive success?

Inga species are characterised by generalist or mixed pollination system. However, this feature does not enhance reproductive rates in species with very low fruit set under natural conditions. Some ecological and genetic factors are associated with this feature, and to test the effect of massive visits on pollination success in Inga subnuda subsp. luschnathiana, we studied the efficacy of polyads deposited on stigmas of flowers isolated from visitors and polyads exposed to visitors. The proportion of polyads fixed in stigmas decreased after exposure to visitors (24 h) in comparison to stigmas isolated from visitors (hummingbirds, bees, wasps, hawkmoths and bats), and fruit set was very low. Furthermore, nectar production, sugar composition and other floral biology traits were evaluated. Increased nectar production, sugar availability and sucrose dominance during the night indicates adaptation to nocturnal visitors and supports their role as main pollinators; although the brush‐flower morphology, time of anthesis, nectar dynamics and chemical composition also allow daytime visitors. Thus the species is an important resource for a diverse group of floral visitors. We conclude that excess visits (diurnal and nocturnal) are responsible for the decrease in fixed polyads in stigmas of I. subnuda subsp. luschnathiana flowers, thus contributing, with others factors, to its low fruit set. Therefore, the generalist pollination system does not result in reproductive advantages because the low fruit set in natural conditions could be the result of a negative effect of visitors/pollinators.     

Philibertia (Apocynaceae: Asclepiadoideae) — additional notes and three new species for Bolivia

Summary  Three new species of Philibertia (Apocynaceae: Asclepiadoideae), P. alba Goyder, P. urceolata Goyder and P. zongoensis Goyder are described from Bolivia and illustrated. Preliminary conservation assessments are made for these species. New records are reported for two further species.     

Systematic implications of the flavonoids and chromosomes ofFlyriella (Compositae—Eupatorieae)

The leaves of five species ofFlyriella were found to contain from one to four glycosides of quercetin and its 4- and 7,4-methyl ethers. These patterns are distinct from those observed for more than seventy species ofBrickellia and support morphological and chromosomal data which indicate thatFlyriella should not be treated as an element ofBrickellia. Alternative treatments are briefly considered.     

The influence of mist and rain on the reproductive success of the Blue Swallow Hirundo atrocaerulea (1995–96)

Evans, S.W. & Bouwman, H. 2000. The influence of mist and rain on the reproductive success of the Blue Swallow Hirundo atrocaerulea. Ostrich 71 (1 & 2): 83–86.

Due to the locality of the Blue Swallow Natural Heritage Site it appears that the birds have no alternative but to forage in the mist. The birds cease foraging as soon as it starts raining. Eight eggs failed to hatch after a week in which rainfall occurred for 60% and mist for 85% of the time. Later in the season a further six eggs failed to hatch after a week in which rainfall occurred for 18% and mist occurred for 65% of the time. The only mortality of two nestling Blue Swallows occurred at the end of a week in which rainfall occurred for 65% and mist was recorded for 85% of the time. At a nest containing three eggs, during the presence of mist (16°C wet/19°C dry) the female Blue Swallow spent a mean of 52 min h^?1 foraging and therefore only a mean of 8 min h^?1 incubating. In the absence of mist (21°C wet/26°C dry) the female Blue Swallow spent a mean of 25 min h^?1 foraging and therefore 35 min h^?1 incubating. Rainfall did not occur during the monitoring of this nest. It appears that the mist greatly reduces the foraging efficiency of the birds and that rainfall duration impacts on the birds by limiting the available foraging time.     

Storage and reproductive strategy of the carpet-shell clam,Ruditapes decussatus in the Gulf of Gabès (Tunisia)

In this study, the variations of glycogen concentrations in Ruditapes decussatus from Sfax coasts (Tunisia) were described in relation to the reproductive cycle. Separate analyses were made of gonad, adductor muscle and ‘remainder’. The timing of gametogenic development and spawning was evaluated using qualitative histology associated with image analysis including (1) the estimation of the gonadal occupation index (GOI), (2) surface area occupied by reserve tissues and (3) variation in oocyte diameter. The reproductive cycle of R. decussatus exhibited partial asynchronization between sexes, the major difference being observed in the duration of the spawning period. Contrary to previous studies, we observed continuous partial spawning (e.g. 50% of ripe oocytes still subsisted at the partial spawning stage). During the gametogenic cycle, GOI varied significantly in males and in females. Most oocytes were ripe and ready to spawn when their diameter reached or exceeded 45?µm. The expansion of gonad was inversely proportional to the development of the foot tissue. Glycogen concentration showed significant temporal variations between tissues, indicating the clear differentiation in energy storage between the clam organs.     

Flowering dynamics and pollination system of the sedge Rhynchospora ciliata (Vahl) Kükenth (Cyperaceae): does ambophily enhance its reproductive success?

Cyperaceae are characteristically anemophilous, but there are some reports of species re‐adapted to entomophily, such as Rhynchospora ciliata. Our objective was to investigate: (1) the distribution pattern of flowers in inflorescences of Rhynchospora ciliata; (2) the dynamics of its anthesis; and (3) whether R. ciliata is pollinated by bees, by wind or by both. Additionally, we tested the hypotheses: (i) the hypsophylls and/or anthers attract pollinators, and (ii) biotic vectors enhance the reproductive success of R. ciliata. We analysed floral biology, dynamics of anthesis, frequency and behaviour of insects visiting flowers; we also carried out experiments on flower attractiveness, pollination by wind and reproductive success. Rhynchospora ciliata has flowers with anemophilous attributes, including anthers exposed during anthesis; however, the anthers (here considered a mixed trait) together with the white hypsophylls can be considered as attributes that favour entomophily. Both wind and four species of bee were considered as pollen vectors of R. ciliata. Through flower attractiveness tests, we observed that the hypsophylls do not affect the frequency of pollinating bees and that the absence of exposed anthers affects the average number of visits, probably because pollen is the only floral resource. Reproductive tests indicate that R. ciliata is self‐incompatible and that ambophily enhances its reproductive success.     

A longstanding entomological problem finally solved? Head morphology of Nannochorista (Mecoptera,Insecta) and possible phylogenetic implications

External and internal head structures of Nannochorista species were examined and described in detail. The characters are discussed with regard to their functional and phylogenetic implications. The structure of the mouthparts indicates that adults of Nannochorista feed on fluids. The loss of the mandibular muscles and the precerebral pharyngeal dilators are presumptive autapomorphies of the genus. A possible clade comprising Nannomecoptera, Siphonaptera and Diptera is supported by the presence of a labral food channel, the absence of the galea, a sheath for the paired mouthparts formed by the labium, very strongly developed labial palp muscles and cibarial dilators, and the presence of a well‐defined postcerebral pharyngeal pumping chamber. Closer affinities of Nannomecoptera with Diptera are suggested by the presence of a unique sensorial groove on the third maxillary palpomere. Further potential synapomorphies are the presence of a frontal apodeme and a primarily lamelliform mandible without teeth. The presence of a salivary channel on the laciniae and a subdivided labrum are shared derived features of Nannochorista and Siphonaptera. A derived condition present in Mecoptera including Boreidae but excluding Nannochoristidae is the secretion with a strongly developed intrinsic muscle of the salivary duct. The loss of the lateral labral retractor, the cranial muscle of the cardo, and of two of the three premental retractors, and the absence of transverse epipharyngeal muscles are potential autapomorphies of Antliophora. The formation of a maxillolabial complex is a possible synapomorphy of Hymenoptera and Mecopterida.     

Effect of liquid culture on the growth and development of miniature rose (Rosa chinensis Jacq. ‘Minima’)

The growth of miniature rose (Rosa chinensis Jacq. Minima) shoots cultured on liquid medium was greater relative to those cultured on two-phase (solid + liquid) medium or solid medium alone. Shoot multiplication ratio (number of multiple shoots per explant per subculture) on liquid medium was higher with 17.8–26.6 M 6-benzyladenine at compared to that at 0–8.9 M. Shoots grown on 30 ml or more of liquid medium had a higher multiplication ratio than those grown on 10 or 20 ml. The growth and multiplication ratio increased when the culture period was extended from 3 to 6 weeks, although plantlets began to exhibit some chlorosis by the 6^th week. These conditions were maintained over four subcultures for cultivars Baby Katie, Lavender Jewel, Red Sunblaze and Royal Sunblaze, with no significant change in multiplication ratio over time.Abbreviations BA 6-benzyladenine - NAA 1-naphthaleneacetic acid     

The Bainbridgia — Dualipora association (Cnidaria,Tabulata): palaeogeographical and palaeoecological implications

The genera Bainbridgia and Dualipora (Cnidaria, Tabulata) have been found together in 4 localities of Early Devonian age (Missouri, USA; Tafilalt, Morocco; Barrandian, Czechoslovakia; Hörre, Germany). Several isolated occurrences of each are also reported. The palaeogeographical and palaeoecological implications of this association are discussed.     

Localized hybridization betweenSaxifraga aizoides andS. mutata (Saxifragaceae): Reproductive ecology ofS. ×hausmannii and implications for conservation

The roles of hybridization and of introgression as potential threats to small native populations are current topics of interest both for evolutionary and conservation biologists. Two factors are mainly involved in the threat to natural populations through hybridization: demographic swamping and genetic assimilation. At prealpine locations, twoSaxifraga species with very different growth habits,S. aizoides andS. mutata, form a hybrid,S. ×hausmannii. Investigation of meiosis and crossing experiments revealed regular pairing of the 26 chromosomes and development of viable seeds in the hybrid. Analysis of hybrid indices, GPI allozymes, germination experiments and seed sizes showed an intermediate position ofS. ×hausmannii between its parental species.Saxifraga aizoides andS. mutata were separated by only weak hybridization barriers. Introgression occurred mostly in direction ofS. aizoides, although to a small amount. There is no actual threat to small, prealpine populations ofS. aizoides andS. mutata through interspecific gene flow because hybridization is localized, resulting from different flowering phenologies of the parental species and from the narrow habitat requirements ofS. ×hausmannii, which are similar to those ofS. aizoides. The major threat to local, prealpine populations ofS. aizoides andS. mutata is the loss of landscape dynamics. Both species need open erosion slopes for successful recruitment.     

Better common than rare? Effects of low reproductive success,scarce pollinator visits and interspecific gene flow in threatened and common species of Tibouchina (Melastomataceae)

The reasons for plant rarity have been the focus of many studies, especially because rare species are more prone to extinction than common species. Under the same habitat conditions, rare plants are expected to attract fewer flower visitors and to show some limitation in their reproductive success. Here, using one of the most emblematic Neotropical plant genus (Tibouchina) we tested whether narrow endemic and threatened species in Ecuador have a lower reproductive success or are visited by fewer pollinators than common species, in 13 populations monitored from 2011 to 2013. We also assessed whether interspecific gene flow could be considered a threat to the rare species. However, contrary to expectations, we found that few pollinators visited the flowers, independently of species rarity. Natural outcross pollinations were always very low in all small‐size populations, leading to high levels of pollen limitation. Interspecific crossing experiments also revealed weak reproductive barriers in some species. This study reveals that both narrow and common species of Tibouchina have similar reproductive and pollinator patterns in Ecuador and, therefore, other causes of the rarity of these species should be considered.