Mapping the mutation causing lens luxation in several terrier breeds

Primary lens luxation (PLL), a painful and blinding inherited condition, is common in several breeds of terrier. Here we have examined the Veterinary Medical Database of patient encounters and Canine Eye Registration Foundation (CERF) cases records for the last 10 years and found the diagnosis recorded in 85 breeds. We have performed association analysis using a genome-wide microsatellite screen to map mutations underlying the condition in miniature bull terriers and Lancashire heelers. These studies show microsatellite alleles in disequilibrium with disease status with highest support in a 6.3-Mbp region in the central part of chromosome 3 (-log P(max) = 6.4). The same region also shows an association to the disease in Tibetan terriers. Tight junction protein-1 (TJP1) is a positional candidate to contain the PLL mutation. All recognized exons and splice junctions of TJP1 have been sequenced from affected, obligate carrier and control Lancashire heeler dogs. Several polymorphisms have been found, but these are not likely to cause the disease.     

Quantification of global transcription patterns in prokaryotes using spotted microarrays

We describe an analysis, applicable to any spotted microarray dataset produced using genomic DNA as a reference, that quantifies prokaryotic levels of mRNA on a genome-wide scale. Applying this to Mycobacterium tuberculosis, we validate the technique, show a correlation between level of expression and biological importance, define the complement of invariant genes and analyze absolute levels of expression by functional class to develop ways of understanding an organism's biology without comparison to another growth condition.     

Effect of coculturing canine notochordal,nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration

IntroductionEarly degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs.MethodsCanine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture.ResultsNCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa.ConclusionsNo regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-015-0569-6) contains supplementary material, which is available to authorized users.     

A new approach to determining environmental flow requirements: Sustaining the natural values of floodplains of the southern Murray‐Darling Basin

Summary Large overbank flood events play an important role in maintaining large‐scale ecological processes and connectivity along and across the floodplains and between the rivers and their floodplains in the southern Murray‐Darling Basin. However, the regulation of rivers means that extensive overbank flooding can only occur in the rare circumstance of extreme flood events. Recent environmental water allocations have focussed on the largest floodplain blocks (‘icon’ sites) and a small set of specific values (e.g. colonial nesting waterbirds), as well as on trialling fine‐scale manipulation of infrastructure (e.g. pumping) to water relatively small areas. There has been no comprehensive systematic assessment of the entire floodplain and its wider set of flood‐dependent natural assets (such as ecosystems and species; herein referred to as ‘natural values’) to maximise the effectiveness of environmental water use and to catalogue values likely to be lost. This paper describes an assessment of some 220 000 ha found to support flood‐dependent natural values in Victoria. We mapped the geographic distribution and estimated components of the flooding requirements (natural flooding frequency, and maximum period without flooding and minimum duration of each flooding event before significant deterioration) for each natural value. Using an example of one stretch of the River Murray, we show how the resultant spatial data can be used with floodplain inundation modelling to compare the outcomes of real or planned environmental watering events; potentially providing tools for management agencies to conserve a wider range of floodplain values than is currently the case. That is, water managers and the public can see what ecosystems and threatened species are intended to be maintained by environmental watering and what values are intended to be abandoned across the whole floodplain, rather than just seeing the small subset of values and ‘icon’ sites that are intended to be maintained. Examples are provided to illustrate how information about the location, water requirements and extent covered by potential floods for specific values can be used to build adaptive watering strategies for areas as large as the whole floodplain.     

Replication of extended lifespan phenotype in mice with deletion of insulin receptor substrate 1

We previously reported that global deletion of insulin receptor substrate protein 1 (Irs1) extends lifespan and increases resistance to several age-related pathologies in female mice. However, no effect on lifespan was observed in male Irs1 null mice. We suggested at the time that the lack of any effect in males might have been due to a sample size issue. While such lifespan studies are essential to our understanding of the aging process, they are generally based on survival curves derived from single experiments, primarily due to time and economic constraints. Consequently, the robustness of such findings as a basis for further investigation has been questioned. We have therefore measured lifespan in a second, separate cohort of Irs1 null female mice, and show that, consistent with our previous finding, global deletion of Irs1 significantly extends lifespan in female mice. In addition, an augmented and completed study demonstrates lifespan extension in male Irs1 null mice. Therefore, we show that reduced IRS1-dependent signalling is a robust mechanism through which mammalian lifespan can be modulated.