Reproduction Is Associated with a Tissue-Dependent Reduction of Oxidative Stress in Eusocial Female Damaraland Mole-Rats (Fukomys damarensis)

Oxidative stress has been implicated as both a physiological cost of reproduction and a driving force on an animal''s lifespan. Since increased reproductive effort is generally linked with a reduction in survival, it has been proposed that oxidative stress may influence this relationship. Support for this hypothesis is inconsistent, but this may, in part, be due to the type of tissues that have been analyzed. In Damaraland mole-rats the sole reproducing female in the colony is also the longest lived. Therefore, if oxidative stress does impact the trade-off between reproduction and survival in general, this species may possess some form of enhanced defense. We assessed this relationship by comparing markers of oxidative damage (malondialdehyde, MDA; protein carbonyls, PC) and antioxidants (total antioxidant capacity, TAC; superoxide dismutase, SOD) in various tissues including plasma, erythrocytes, heart, liver, kidney and skeletal muscle between wild-caught reproductive and non-reproductive female Damaraland mole-rats. Reproductive females exhibited significantly lower levels of PC across all tissues, and lower levels of MDA in heart, kidney and liver relative to non-reproductive females. Levels of TAC and SOD did not differ significantly according to reproductive state. The reduction in oxidative damage in breeding females may be attributable to the unusual social structure of this species, as similar relationships have been observed between reproductive and non-reproductive eusocial insects.     

Early Detection of Motor Dysfunction in the SOD1G93A Mouse Model of Amyotrophic Lateral Sclerosis (ALS) Using Home Cage Running Wheels

The SOD1^G93A mouse has been used since 1994 for preclinical testing in amyotrophic lateral sclerosis (ALS). Despite recent genetic advances in our understanding of ALS, transgenic mice expressing mutant SOD1 remain the best available, and most widely used, vertebrate model of the disease. We previously described an optimised and rapid approach for preclinical studies in the SOD1^G93A mouse. Here we describe improvements to this approach using home cage running wheels to obtain daily measurements of motor function, with minimal intervention. We show that home cage running wheels detect reductions in motor function at a similar time to the rotarod test, and that the data obtained are less variable allowing the use of smaller groups of animals to obtain satisfactory results. This approach refines use of the SOD1^G93A model, and reduces the number of animals undergoing procedures of substantial severity, two central principles of the 3Rs (replacement, reduction and refinement of animal use in research). The small group sizes and rapid timescales enable affordable large-scale therapeutic pre-screening in the SOD1^G93A mouse, as well as rapid validation of published positive effects in a second laboratory, one of the major stumbling blocks in ALS preclinical therapy development.     

Nuclear DNA Amounts in Angiosperms and their Modern Uses--807 New Estimates

The DNA amount in the unreplicated haploid nucleus of an organismis known as its C-value. C-values differ about 1000-fold amongangiosperms and are characteristic of taxa. The data are usedin many biological fields, so they should be easily available.Values for 2802 angiosperm species (1%) were estimated during1950–1997, and five collected lists of C-values were publishedfor reference purposes during 1976–1997. Numbers of newangiosperm C-values published recently remained high, necessitatinga further supplementary list. This paper lists DNA C-valuesfor 807 angiosperm species from 70 original sources, including520 (75.2%) from sources published after 1996, and 691 for speciesnot included in any of the previous five lists. There is a continuingneed to estimate accurate DNA C-values for plant taxa, as shownin a workshop on this biodiversity topic sponsored by Annalsof Botany and held at Kew in 1997. Its key aim was to identifymajor gaps in our knowledge of plant DNA amounts and to recommendtargets and priorities for new work to fill them. A target ofestimating first C-values for the next 1% of angiosperm speciesin 5 years was set. The proportion of such C-values in the presentwork (85.6%) is very high; and the number being published (approx.220 per annum) has never been exceeded. In 1997, C-values werestill unknown for most (68%) families, so a target of completecoverage was set. This paper includes first C-values for 12families, but as less than 2% of such values listed here targetednew families, the need to improve familial representation remains.Copyright 2000 Annals of Botany Company Angiosperm DNA amounts, DNA C-values, nuclear genome sizes, plant DNA database     

Quantification of the Role of Acclimation Temperature in Temperature Tolerance of Fishes

The relative effect of acclimation temperature on temperature tolerance was estimated from a geometrical partitioning of the temperature tolerance polygon of a fish species into three distinct zones relative to four key tolerance temperatures. This approach yields a middle tolerance zone which is independent of acclimation temperature bounded by upper and lower acclimation dependent zones. Acclimation dependent and independent temperature tolerance zones can be quantified by either areal or linear methods. Both methods were applied to quantify the effect of acclimation temperature in 21 species of temperate fishes for which temperature tolerance polygons were available. Temperature tolerance polygon areas of these 21 species ranged from 468 to 1380°C² and are linearly related (r ²=0.93, p<0.001) to ultimate incipient upper lethal temperatures. Although areal and linear partitioning methods yielded similar acclimation independent and dependent tolerances, estimates from the areal method incorporates additional information concerning the shape of the temperature tolerance polygon, in particular lower and upper lethal temperature plateaus. Mean combined acclimation dependent and independent tolerance areas of these 21 species were not different, indicating that acclimation effectively doubles the temperature tolerance polygon. Mean lower acclimation dependent area was nearly three times greater than mean upper acclimation dependent area, suggesting that acclimation plays a larger role in tolerance of low rather than high temperatures. Among these 21 species, temperature tolerance of brook charr and sheepshead minnow were the least and most affected by acclimation temperature, respectively.     

Biodiversity of Asian rice gall midge (Orseolia oryzae Wood Mason) from five countries examined by AFLP analysis.

Amplified fragment length polymorphism (AFLP) analysis was used to assess the biodiversity of one of the most important dipteran pests of cereals, the Asian rice gall midge (Orseolia oryzae Wood Mason). Larvae and pupae were collected at 15 locations in five Asian countries and preserved in 95% ethanol for storage, shipment, and DNA extraction using cetyltrimethylammonium bromide (CTAB). Although only approximately 1 microg of DNA was extracted from a single pupa or larva, the use of several AFLP primers in various combinations meant that this amount of DNA was sufficient to allow many DNA fingerprints to be made per individual. Fingerprints were sufficiently reproducible, especially during selective amplification, to allow the genetic diversity within a field population to be characterized. Extraction of DNA from a pool of 20 insects yielded AFLP fingerprints in which variation among individuals was sacrificed in favor of detecting differences among populations. For each location, pooled DNA was amplified with three primer pairs. A total of 261 distinct AFLP bands were identified for the 45 fingerprints. Cluster analysis, performed by the unweighted pair-group method (UPGMA), separated the populations into two distinct groups. Group I included two populations from Guangdong province of southern China and one each from Laos and Imphal in northeastern India, while group II was comprised of eleven populations from elsewhere in India (Assam, Orissa, Madhya Pradesh, Andhra Pradesh, and Kerala) and from Nepal and Sri Lanka. AFLP analysis provided insight into the origins of gall midge biotypes. In 1992, the prevailing biotype in Imphal changed from Indian biotype 3 to a new biotype 3M. Our data show that biotype 3M belongs to group I and did not arise by a recent mutation from biotype 3, which belongs to group II. By contrast, Indian biotypes 2 and 4 are likely to have diverged through recent mutation and selection, as are Chinese biotypes 1 and 4. The almost simultaneous emergence of new biotypes in Kerala and Sri Lanka during 1985-1988 was most probably coincidental, because these biotypes are not closely related. AFLP fingerprints were also able to detect sexual dimorphism in the DNA of adult gall midges and to distinguish gall midge from its major parasite Platygaster oryzae.     

P2X (purinergic) receptor redistribution in rabbit aorta following injury to endothelial cells and cholesterol feeding

The redistribution of purinergic P2X receptor subunits (P2X₁ to P2X₇) within the rabbit aorta wall three weeks after endothelial balloon injury/cholesterol feeding was examined. P2X₁ receptor cluster density was elevated in the media following balloon injury/cholesterol feeding by about 30% and these clusters appeared on smooth muscle cells throughout the greatly expanded neointima but they did not change significantly on the endothelial cells following balloon injury. P2X₄ clusters were found in high density throughout the media and in very high density in the enlarged neointima following balloon injury, particularly on the endothelial cells where the density increased about 10-fold after balloon injury. P2X₅ clusters were found in high density in the media of normal aorta but with little change following balloon injury. P2X₃, P2X₆ and P2X₇ cluster density was low in normal aorta and remained unchanged following balloon injury. All receptor subunits were found on endothelial cells. It is suggested that the release of ATP from damaged endothelial cells and from smooth muscle cells sufficient to activate P2X₄ receptors may contribute to neointimal proliferation.     

Changes in the distribution of different subtypes of P2X receptor clusters on smooth muscle cells in relation to nerve varicosities in the pregnant rat urinary bladder

Clusters of purinergic receptor subunits, about 1 μm diameter, are found on the smooth muscle cell membrane beneath junctional varicosities in the detrusor muscle of the rat urinary bladder. We have examined the extent of redistribution of the six different subunit clusters, P2X₁ to P2X₆, with respect to junctional varicosities during pregnancy, as it is known that the detrusor muscle undergoes changes in purinergic innervation during this period. Before pregnancy, clusters at junctional varicosities are principally composed of the subtypes P2X₁, P2X₂, P2X₃ and P2X₅. However this subtype distribution changes dramatically during pregnancy, such that by day 14 of pregnancy, the extent of P2X₁, P2X₂, P2X₃ and P2X₅ junctional clusters has decreased by more than 80% whereas the extent of P2X₄ and P2X₆ junctional clusters has increased by more than 80%. These changes were confirmed with Western blots for different subtypes. It is suggested that the changes in the purinergic innervation of the detrusor muscle during pregnancy reflect changes in the P2X subtypes found on the smooth muscle membrane beneath junctional varicosities.