Barriers to cross-fertilization between populations of a widely dispersed Polychaete species are unlikely to have arisen through gametic compatibility arms-races

Although there are theoretical reasons to suspect that gametic incompatibility may develop readily among populations of broadcast spawning marine invertebrates, there have been very few studies documenting geographic patterns of interpopulation incompatibility for any species. To address this we determined how successfully individuals of the intertidal serpulid polychaete, Galeolaria caespitosa, can cross-fertilize within and among populations from across temperate Australia. Fertilization assays revealed asymmetrical differences between very distantly located populations from different coasts, with near-complete incompatibility between eggs from Sydney with sperm from Adelaide, but the reverse cross (Adelaide eggs, Sydney sperm) was reasonably compatible. Although that pattern was congruent with a clear difference in Cytochrome B sequences between worms on the south and east coasts of Australia, we also detected some indication of interpopulation incompatibility within the genetic grouping on east coast, between two populations separated by only 220 km. We then assessed whether commonly proposed gametic compatibility arms-races could account for these patterns. Our results suggest reduced gametic compatibility may reduce a female's maximum fertilization potential, resulting in a cost to this potential mechanism for reducing polyspermy. Consequently, the apparently rapid development of reproductive barriers here seems unlikely to have been driven by arms-races involving sexual conflict over fertilization rate.

Male and female effects on fertilization success and offspring viability in the Peron's tree frog, Litoria peronii

There is increasing theoretical and empirical evidence that genetic compatibility among partners is an important determinant of fertilization success and offspring viability. In amphibians, females often actively choose partners from among a variety of males and polyandry is common. Genetic compatibility among partners may therefore be an important determinant of fertilization success and offspring viability in some amphibians. Amphibians also show some of the highest levels of genetic differentiation among neighbouring populations known in vertebrates, and as such, populations may have evolved different co-adapted gene complexes. This means that offspring from among-population crosses may have reduced fitness. It is therefore essential to understand to what extent crossings between and within populations may interfere with successful fertilization and offspring viability. Here, we test whether crossing individuals within and between two different populations of the Australian Peron's tree frog (Litoria peronii) using artificial fertilizations affect fertilization success and offspring viability. Fertilization success per se is strongly influenced by male identity, which is likely to depend at least to some extent on the experimental procedure (e.g. resulting in variation in sperm number per ejaculate), whereas there was no fertilization effect of female identity. More importantly, male and female identity, independently of each other, explained significant variation in offspring viability, whereas no such effect could be linked to population of origin. Thus, our experiments suggest that crossing populations may not always be the most significant factor affecting fertilization success or offspring viability, but may be more influenced by the genetic quality or the genetic compatibility of partners.

Female choice for males with greater fertilization success in the Swedish Moor Frog, Rana arvalis

Background: Studies of mate choice in anuran amphibians have shown female preference for a wide range of male traits despite females gaining no direct resources from males (i.e. non-resource based mating system). Nevertheless, theoretical and empirical studies have shown that females may still gain indirect genetic benefits from choosing males of higher genetic quality and thereby increase their reproductive success.
Methodology/Principal Findings: We investigated two components of sexual selection in the Moor frog (Rana arvalis), precopulatory female choice between two males of different size (‘large’ vs. ‘small’), and their fertilization success in sperm competition and in isolation. Females’ showed no significant preference for male size (13 small and six large male preferences) but associated preferentially with the male that subsequently was the most successful at fertilizing her eggs in isolation. Siring success of males in competitive fertilizations was unrelated to genetic similarity with the female and we detected no effect of sperm viability on fertilization success. There was, however, a strong positive association between a male’s innate fertilization ability with a female and his siring success in sperm competition. We also detected a strong negative effect of a male’s thumb length on his competitive siring success.
Conclusions/Significance: Our results show that females show no preference for male size but are still able to choose males which have greater fertilization success. Genetic similarity and differences in the proportion of viable sperm within a males ejaculate do not appear to affect siring success. These results could be explained through pre- and/or postcopulatory choice for genetic benefits and suggest that females are able to perceive the genetic quality of males, possibly basing their choice on multiple phenotypic male traits.

Isolation of sperms from the pollen tube of flowering plants during fertilization

Sperm cells have been isolated from pollen tubes growing in style segments of the dicotlyledon Rhododendron macgregoriae and the monocotyledon Gladiolus gandavensis by the in vivo/in vitro method at various stages of fertilization. Pollen tubes emerged from the cut end of the style into agar medium, and more than 95% contained sperm cells. Sperm cells were released from the pollen tubes by osmotic shock or by placing styles in wall-degrading enzymes: 0.5% macerozyme and 1% cellulase. The isolated sperms were ellipsoidal protoplasts of diameter about 2 × 3 micrometers in Gladiolus and about 3 × 4 micrometers in Rhododendron. After isolation, a proportion of the sperm cells occurred in pairs linked at one end by finger-like connections. The pairs of isolated sperms were dimorphic in terms of surface area and volume. By cutting the styles at various positions and times after pollination, the potential exists to detect changes in sperm gene expression associated with fertilization.

Use of pineapple juice for elimination of egg stickiness of common carp (Cyprinus carpio L.)

Four treatments including three different concentrations of pineapple solution (I %, 3%, 5%) and the traditional method (salt/urea/tannin solutions) were used to eliminate egg stickiness of common carp. With the traditional method, fertilization and hatching rates were 74.5% &plusmn; 1.2 and 70.2% &plusmn; 1.1, respectively. The highest fertilization rate (89.3% &plusmn; 0.7) and hatching rate (86.6% &plusmn; 1.4) were found with treatment of 1% pineapple juice solution and was significantly different from those obtained with other treatments (P<0.05). Using pineapple juice for desticking common carp eggs also reduced the time of egg handling from almost one hour required with the traditional method, to about three minutes, as well as increasing fertilization and hatching rate under hatchery conditions.

Scallop size does not predict amount or rate of induced sperm release

Within populations of broadcast spawning marine invertebrates such as scallops, larger animals typically have larger gonads. Presumably, this means those larger males have more sperm to release than small males. However, there has never been a direct test of whether larger males actually release more sperm, at a higher rate, during spawning. To address this, we compared the allometry of induced sperm release with that of reproductive investment (gonad weight) in ripe males of 2 species of scallops, Chlamys bifrons and Chlamys asperrima. We did not find that larger scallops released more sperm or released it faster than small scallops, and were able to reject the hypothesis that instantaneous sperm release was related to body size in the same way as gonad weight. Consequently, we speculate that if larger broadcast spawning males do release more sperm, they may do so by spawning on more occasions within a reproductive season.

Sexual conflict and polyspermy under sperm-limited conditions : in situ evidence from field simulations with the free-spawning marine echinoid Evechinus chloroticus

For free‐spawning organisms that release gametes into the sea, sperm limitation (too few sperm to fertilize all eggs) is a major factor limiting reproductive success. Given such circumstances, the presence of several mechanisms to prevent polyspermy (too many sperm) may seem paradoxical; however, a growing body of data suggests that natural fertilization levels, though variable, can routinely be high. Under such conditions, polyspermy is much more likely. The tension between sperm limitation and polyspermy represents sexual conflict because males, in competing to fertilize as many eggs as possible, can impose lethal costs on eggs if multiple sperm gain entry. Here we present data for a marine invertebrate indicating high levels of polyspermy under sperm‐limited conditions. When the sea urchin Evechinus chloroticus was induced to spawn in situ, mean rates of polyspermy were 17.3% &plusmn;3.4%, and polyspermy was recorded at rates as high as 62.7%. Polyspermy was nearly always present, even when fertilization rates were <50%, confirming predictions that it should be present under sperm‐limited conditions. Both sperm limitation and polyspermy imposed substantial reproductive costs, and we conclude that both sexual conflict related to polyspermy and sperm limitation have been simultaneous strong selective forces shaping the evolution of reproductive traits in the sea.

Sperm environment affects offspring quality in broadcast spawning marine invertebrates

The provisioning of offspring can have far-reaching consequences for later life in a wide range of organisms and generally this provisioning is thought to be under maternal influence or control. In experiments with a broadcast-spawning ascidian, we found that the size of offspring was determined by egg size and the abundance of sperm present during fertilization. Larger eggs were fertilized at low sperm concentrations, whilst smaller eggs were successfully fertilized at high sperm concentrations. These differences in fertilized egg size resulted in differences in the development rate, hatching success and mean size of the subsequent larvae. Our results suggest that, in contrast to females that reproduce by other mating systems, free-spawning mothers lack some control over the provisioning of offspring. Furthermore, because males can alter the sperm environment, they can exert paternal (non-genetic) control over key offspring characteristics.

The role of jak2a in zebrafish hematopoiesis

Janus kinase 2 (Jak2) transduces signals from hematopoietic cytokines, and a gain-of-function mutation (Jak2617V>F) is associated with myeloproliferative diseases, particularly polycythemia vera. In this study, we examined the role of jak2a in zebrafish embryos in knock-down and overexpression studies using morpholinos (MOs) targeting the 5' untranslated region (UTR) (jak2aUTR-MO) and splice-site junction (jak2aSS-MO) of jak2a, a Jak inhibitor AG490 and a constitutive-active form of jak2a (jak2aca). At 18 and 24 hours after fertilization (hpf), jak2a is expressed predominantly in the intermediate cell mass (ICM; site of primitive hematopoiesis) of wild-type and chordin morphant embryos (characterized by expansion of ICM). Both jak2a MOs and AG490 reduced gata1+ (erythroid) cells in Tg(gata1:GFP) embryos, signal transducer and activation of transcription 5 (stat5) phosphorylation, and gene expression associated with early progenitors (scl and lmo2) and erythroid (gata1, he1 and ßhe1) and myeloid (spi1 [early] and mpo [late]) lineages. The chordin morphant is associated with increased stat5 phosphorylation, and both jak2a MOs and treatment with AG490 significantly ameliorated ICM expansion and hematopoietic gene up-regulation in these embryos. Injection of plasmid encoding jak2aca significantly increased erythropoiesis and expression of gata1, he1 and ßhe1, spi1, mpo, and l-plastin. In conclusion, zebrafish jak2a is involved in primitive hematopoiesis under normal and deregulated conditions.

Characterization of the zebrafish matrix metalloproteinase 9 gene and its developmental expression pattern

Members of the matrix metalloproteinase (MMP) family are important for the remodeling of the extracellular matrix in a number of biological processes including a variety of immune responses. Two members of the family, MMP2 and MMP9, are highly expressed in specific myeloid cell populations in which they play a role in the innate immune response. To further expand the repertoire of molecular reagents available to study zebrafish myeloid cell development, the matrix metalloproteinase 9 (mmp9) gene from this organism has been identified and characterized. The encoded protein is 680 amino acids with high homology to known MMP9 proteins, particularly those of other teleost fish. Maternal transcripts of mmp9 are deposited in the oocyte and dispersed throughout the early embryo. These are replaced by specific zygotic transcripts in the notochord from 12 h post fertilization (hpf) and also transiently in the anterior mesoderm from 14 to 16 h post fertilization. From 24 h post fertilization, mmp9 expression was detected in a population of circulating white blood cells that are distinct from macrophages, and which migrate to the site of trauma, and so likely represent zebrafish heterophils. In the adult, mmp9 expression was most prominent in the splenic cords, a site occupied by mature myeloid cells in other species. These results suggest that mmp9 will serve as a useful marker of mature myeloid cells in the zebrafish.

Genomic analyses and cloning of novel chicken natriuretic peptide genes reveal new insights into natriuretic peptide evolution

The natriuretic peptide (NP) family consists of multiple subtypes in teleosts, including atrial, B-type, ventricular, and C-type NPs (ANP, BNP, VNP, CNP-1&ndash;4, respectively), but only ANP, BNP, CNP-3, and CNP-4 have been identified in tetrapods. As part of understanding the molecular evolution of NPs in the tetrapod lineage, we identified NP genes in the chicken genome. Previously, only BNP and CNP-3 have been identified in birds, but we characterized two new chicken NP genes by cDNA cloning, synteny and phylogenetic analyses. One gene is an orthologue of CNP-1, which has only ever been reported in teleostei and bichir. The second gene could not be assigned to a particular NP subtype because of high sequence divergence and was named renal NP (RNP) due to its predominant expression in the kidney. CNP-1 mRNA was only detected in brain, while CNP-3 mRNA was expressed in kidney, heart, and brain. In the developing embryo, BNP and RNP transcripts were most abundant 24 h post-fertilization, while CNP mRNA increased in a stage-dependant manner. Synthetic chicken RNP stimulated an increase in cGMP production above basal level in chicken kidney membrane preparations and caused a potent dose-dependant vasodilation of pre-constricted dorsal aortic rings. From conserved chromosomal synteny, we propose that the CNP-4 and ANP genes have been lost in chicken, and that RNP may have evolved from a VNP-like gene. Furthermore, we have demonstrated for the first time that CNP-1 is retained in the tetrapod lineage.

A new broad composite model of regional economic development as a framework for research

The mechanism of regional economic development has been studied extensively by economists, geographers, town planners and other academics. The existing wide varieties of theories into regional economic development are insufficient on their own in explaining how a region can develop and prosper. Each theory has evaluated a few facets of regional economic development. Research from these different perspectives is narrow and prevents any cross-fertilization of research from all these diverse
theories.

Recognition of multiple factors affecting the development process has led the author to create a new broad composite model of regional economic development. The paper first sets out to describe and explain this broad composite model. Each of the components of this new model draws heavily upon seminal work in the field. This model proposes three rings. Each ring is at a different level of abstraction. The determinants of development described in each ring can influence each and every other determinant of development shown in the three ring structure. This model recognises that development in any centre, regional or urban, nascent or established is a composite end result of the complex interplay of all the determinants of development.

The paper then goes on to show how this model can provide a broad holistic approach to regional economic development that can assist researchers in their attempts to understand and link the various theories of regional economic development.

Morphologic and functional characterization of granulocytes and macrophages in embryonic and adult zebrafish.

The zebrafish is a useful model organism for developmental and genetic studies. The morphology and function of zebrafish myeloid cells were characterized. Adult zebrafish contain 2 distinct granulocytes, a heterophil and a rarer eosinophil, both of which circulate and are generated in the kidney, the adult hematopoietic organ. Heterophils show strong histochemical myeloperoxidasic activity, although weaker peroxidase activity was observed under some conditions in eosinophils and erythrocytes. Embryonic zebrafish have circulating immature heterophils by 48 hours after fertilization (hpf). A zebrafish myeloperoxidase homologue (myeloid-specific peroxidase; mpx) was isolated. Phylogenetic analysis suggested it represented a gene ancestral to the mammalian myeloperoxidase gene family. It was expressed in adult granulocytes and in embryos from 18 hpf, first diffusely in the axial intermediate cell mass and then discretely in a dispersed cell population. Comparison of hemoglobinized cell distribution, mpx gene expression, and myeloperoxidase histochemistry in wild-type and mutant embryos confirmed that the latter reliably identified a population of myeloid cells. Studies in embryos after tail transection demonstrated that mpx- and peroxidase-expressing cells were mobile and localized to a site of inflammation, indicating functional capability of these embryonic granulocytes. Embryonic macrophages removed carbon particles from the circulation by phagocytosis. Collectively, these observations have demonstrated the early onset of zebrafish granulopoiesis, have proved that granulocytes circulate by 48 hpf, and have demonstrated the functional activity of embryonic granulocytes and macrophages. These observations will facilitate the application of this genetically tractable organism to the study of myelopoiesis.

Strategic review of enhancements and culture-based fisheries

Enhancements are interventions in the life cycle of common-pool aquatic resources. Enhancement technologies include culture-based fisheries, habitat modifications, fertilization, feeding and elimination of predators/competitors. Enhancements are estimated to yield about two million mt per year, mostly from culture-based fisheries in fresh waters where they account for some 20 percent of capture, or 10 percent of combined capture and culture production. Marine enhancements are still at an experimental stage, but some have reached commercial production. Enhancements use limited external feed and energy inputs, and can provide very high returns for labour and capital input. Moreover, enhancement initiatives can facilitate institutional change and a more active management of aquatic resources, leading to increased productivity, conservation and wider social benefits. Enhancements may help to maintain population abundance, community structure and ecosystem functioning in the face of heavy exploitation and/or environmental degradation. Negative environmental impacts may arise from ecological and genetic interactions between enhanced and wild stocks. Many enhancements have not realised their full potential because of a failure to address specific institutional, technological, management and research requirements emanating from two key characteristics. Firstly, enhancement involves investment in common-pool resources and can only be sustained under institutional arrangements that allow regulation of use and a flow of benefits to those who bear the costs of enhancement. Secondly, interventions are limited to certain aspects of the life cycle of stocks, and outcomes are strongly dependent on natural conditions beyond management control. Hence, management must be adapted to local conditions to be effective, and certain conditions may preclude successful enhancement altogether. Governments have a major role to play in facilitating enhancement initiatives through the establishment of conducive institutional arrangements, appropriate research support, and the management of environmental and other impacts on and from enhancements.

Who's afraid of designer babies?

Australian scientists in Sydney made medical history by creating a life-saving brother for a child with an incurable genetic disease. Features the case of a Tasmanian couple who, with their help, have this life-saving baby, at their third try. The pioneering IVF treatment, which is legal only in N.S.W., hit the news headlines, sparking an ethical storm. Examines the moral and ethical issues that genetic manipulation raises, including the potential uses and misuses of genetic technology. Raises also the spectre of an extreme form of eugenics, as seen in the World War II Nazi push to create a master race through human genetics. Some view eugenics as another form of PGD. Presents the diverse views of eminent ethicists and the Catholic Church, world wide.