Sherralee S. Lukehurst

Functional Topography and Integration of the Contralateral and Ipsilateral Retinocollicular Projections of Ephrin-A−/− Mice

Topographically ordered projections are established by molecular guidance cues and refined by neuronal activity. Retinal input to a primary visual center, the superior colliculus (SC), is bilateral with a dense contralateral projection and a sparse ipsilateral one. Both projections are topographically organized, but in opposing anterior–posterior orientations. This arrangement provides functionally coherent input to each colliculus from the binocular visual field, supporting visual function. When guidance cues involved in contralateral topography (ephrin-As) are absent, crossed retinal ganglion cell (RGC) axons form inappropriate terminations within the SC. However, the organization of the ipsilateral projection relative to the abnormal contralateral input remains unknown, as does the functional capacity of both projections. We show here that in ephrin-A−/− mice, the SC contains an expanded, diffuse ipsilateral projection. Electrophysiological recording demonstrated that topography of visually evoked responses recorded from the contralateral superior colliculus of ephrin-A−/− mice displayed similar functional disorder in all genotypes, contrasting with their different degrees of anatomical disorder. In contrast, ipsilateral responses were retinotopic in ephrin-A2−/− but disorganized in ephrin-A2/A5−/− mice. The lack of integration of binocular input resulted in specific visual deficits, which could be reversed by occlusion of one eye. The discrepancy between anatomical and functional topography in both the ipsilateral and contralateral projections implies suppression of inappropriately located terminals. Moreover, the misalignment of ipsilateral and contralateral visual information in ephrin-A2/A5−/− mice suggests a role for ephrin-As in integrating convergent visual inputs.

Neurite responses to ephrin-A5 modulated by BDNF: Evidence for TrkB-EphA interactions

In the developing visual system, growing retinal ganglion cell (RGC) axons are exposed to multiple guidance and growth factors. Furthermore, the relative levels of these factors are differentially regulated as topography is roughly established and then refined. We have shown that during the establishment of rough topography (P3), growth cones of pure and explanted RGCs treated with combinations of BDNF and ephrin-A5-Fc responded differently than RGCs treated with BDNF or ephrin-A5-Fc alone (p=0.0083). The response to the combined treatment mimicked that of RGCs cultured with ephrin-A5-Fc alone once topography refines. The guidance cue receptors EphA and TrkB were shown to co-localise in RGCs in vitro. Furthermore, EphA and TrkB receptors interacted directly in in vitro binding assays. Our results suggest that the conversion of growth cone responses from collapse to stabilisation as topography refines, occurs as a result of interactions between EphA and TrkB receptors.

Characterisation of a model for retinal neovascularisation. VEGF model Characterisation

Retinal neovascularisation is a major clinical complication of diabetic retinopathy that takes place late in the disease process and constitutes the most damaging phase resulting in loss of vision (Klein et al., 1984). Neovascularisation is defined as the growth of new blood vessels which, in a disease process such as diabetic retinopathy, occurs in abnormal retinal locations. Long term consequences of retinal neovascularisation include the formation of epiretinal membranes and retinal detachment (Smith et al., 1999). In addition, new blood vessels lack a patent blood retinal barrier and exhibit leukostasis presumably resulting in cytotoxic damage (Ishida et al., 2003; Qaum et al., 2001).

Graded ephrin-A2 expression in the developing hamster superior colliculus

During development, ephrin gradients guide retinal ganglion cell axons to their appropriate topographic locations in the superior colliculus (SC). Expression of ephrin-A2, assessed immunohistochemically in the developing hamster SC, revealed a rostrallow to caudalhigh gradient that is most prominent at postnatal days (P)4 and P7 when topography is established. Double-labelling immunohistochemistry for ephrin-A2 and cell specific markers revealed that ephrin-A2 is expressed exclusively by a subset of neurons. The expression pattern has implications for mechanisms underlying establishment of topography during development and following injury.

Isolation and characterization of 13 polymorphic microsatellite loci for the smooth Cherax cainii and hairy marron C. tenuimanus (Decapoda: Parastacidae)

AbstractNumbers of the critically endangered hairy marron Cherex tenuimanus are declining rapidly in its natural habitat, mainly due to competition with smooth marron C. cainii. To determine whether genetic introgression is occurring between the two species we isolated 13 new polymorphic microsatellite markers from C. cainii using 454 shotgun sequencing. The loci were screened for variation in 20 individuals of C. cainii and eight individuals of C. tenuimanus. There was no evidence of linkage disequilibrium between pairs of loci and only one locus deviated significantly from Hardy–Weinberg equilibrium. The number of alleles per locus ranged from two to 11 across both species.

Characterization of microsatellite loci for the Australian sea urchin Heliocidaris erythrogramma

We report 16 polymorphic microsatellite loci from Heliocidaris erythrogramma, a common sea urchin endemic to temperate Australian waters. These microsatellites were tested in a minimum of 30 individuals, which yielded between five and 14 alleles per locus. Expected heterozygosity ranged from 0.52 to 0.92 with four loci deviating from Hardy–Weinberg expectations. These markers are expected to be useful for experimental studies involving paternity analysis and for quantifying population structure in H. erythrogramma across its geographic range.

Characterization of microsatellite loci for the littorine snail Bembicium vittatum.

We describe the isolation and development of 17 polymorphic microsatellite loci for the intertidal snail Bembicium vittatum (Gastropoda: Littorinidae). The loci were tested in 46 individuals from a single population situated near the centre of the species distribution. No evidence of linkage disequilibrium was detected between any pair of loci. However, two loci showed significant departures from Hardy–Weinberg expectations. The number of alleles per locus ranged from two to 15.

Characterisation of microsatellite loci in the groundwater amphipod Chydaekata sp. (Malacostraca: Paramelitidae)

The groundwater fauna (stygofauna) of the Pilbara region of Western Australia is diverse and largely endemic. Extensive mining in the region poses a potential threat to this fragile ecosystem. Knowledge of the dispersal capabilities of groundwater species will aid in managing both fauna and resources. Ten polymorphic microsatellite loci were used to assess variability in 27 specimens of the groundwater amphipod Chydaekata sp. from Marillana Creek in the Pilbara. The number of alleles ranged from 3 to 15, and observed heterozygosity ranged from 0.077 to 0.885. Nine loci were in HWE, and no pairs of loci showed evidence of linkage. The loci will be useful for aiding the management of stygobitic populations in a disturbed landscape, by understanding dispersal patterns and gene flow within and between tributaries.

Genetic diversity of a hitchhiker and prized food source in the Anthropocene: the Asian green mussel Perna viridis (Mollusca, Mytilidae)

Insight into a species’ native and introduced range is essential in understanding the invasion process. Genetic diversity, propagule pressure and environmental conditions all have been recognised as playing a determinant role in invasion success. Here, we aimed to investigate the genetic diversity and population genetic structure (using the COI mtDNA gene region and 22 nDNA microsatellite markers) of the Asian green mussel Perna viridis within its potential native range in Asia and at introduced locations in the USA and the Caribbean. We also analyse genetic data from vessel intercepts and an incursion. By doing so, we aimed to identify genetic signatures that could allow to track vessel samples to their source and provide further insight into potential high-risk invasive populations or areas. Three top hierarchical clusters were identified using the individual-based Bayesian clustering method in STRUCTURE, corresponding to populations in three world regions: (1) USA and Caribbean, (2) India and (3) Southeast Asia. Within Southeast Asia, additional analysis indicate a shallow genetic differentiation of three subgroups consisting of (3a) Thailand, (3b) Taiwan and Hong-Kong, and (3c) a cluster of Singapore–Indonesia samples. Overall, the population structure found in this study suggests that the markers used could be useful in identifying source populations, particularly between the three mains world regions. Most surprisingly however, this study shows that the genetic diversity of samples collected from vessel intercepts and incursions did not differ significantly from established populations in Southeast Asia. In this region, in addition to the high vessel connectivity and number of P. viridis transported, all sampled populations are likely to pose a comparable risk in terms of genetic diversity. The present work represents the most comprehensive population genetic study of P. viridis, and the first to address the potential genetic introduction risk posed by populations of this species. The information and genetic markers in this study constitute a valuable addition to the tools already used to infer on potential high-risk source populations of P. viridis. They should therefore prove useful for biosecurity surveillance and management actions directed at this species.

Isolation and characterisation of 16 polymorphic microsatellite loci for bight redfish, Centroberyx gerrardi (Actinopterygii : Berycidae), and cross-amplification in two other Centroberyx species

Abstract. Bight redfish, Centroberyx gerrardi, is a demersal teleost endemic to continental shelf and upper slope waters of southern Australia. Throughout most of its range, C. gerrardi is targeted by a number of separately managed commercial and recreational fisheries across several jurisdictions. However, it is currently unknown whether stock assessments and management for this shared resource are being conducted at appropriate spatial scales, thereby requiring knowledge of population structure and connectivity. To investigate population structure and connectivity, we developed 16 new polymorphic microsatellite markers using 454 shotgun sequencing. Two to 15 alleles per locus were detected. There was no evidence of linkage disequilibrium between pairs of loci and all loci except one were in Hardy–Weinberg equilibrium. Cross-amplification trials in the congeneric C. australis and C. lineatus revealed that 11 and 16 loci are potentially useful, respectively. However, deviations from Hardy–Weinberg equilibrium and linkage disequilibrium between pairs of loci were detected at several of the 16 markers for C. australis, and therefore the number of markers useful for population genetic analyses with C. lineatus is likely considerably lower than 11.