The thymus and T-cell commitment: the right niche for Notch?

The current dogma is that the thymus is colonized by progenitors that retain the capacity to generate both T cells and B cells, and that intrathymic Notch signalling determines lineage choice so that T cells, rather than B cells, develop in the thymus. However, evidence is now accumulating to indicate that, at least during fetal life, this is not the case. Rather, it now seems that the fetal thymus is colonized by progenitors that have already made the T-cell versus B-cell lineage choice. We propose an alternative role for Notch signalling in the thymus, which is not to mediate this choice but instead to reveal it by supporting further T-cell differentiation in the thymic microenvironment.     

The VNTR in complex disorders: The forgotten polymorphisms? A functional way forward?

In the last few years, research has focused on single nucleotide polymorphisms (SNPs) in the search for underlying genetic aetiology of complex disorders. This has been afforded by the rapid technological advancement to enable the interrogation of hundreds of thousands of SNPs in one assay via microarrays. However SNPs are only one form of genetic variation and in the midst of the Genome-Wide Association Study (GWAS) explosion Variable Number Tandem Repeat (VNTR) polymorphism exploration has seemingly been left behind. This review will argue that VNTR investigations still hold substantial potential for a role in complex disorders via possible functional properties.     

Are any functionally mature cells of medullary phenotype located in the thymus cortex?

Experiments were undertaken to test if thymocytes of "mature" or "medullary" phenotype were restricted to the medullary area of the thymus. A calculation based on direct cell counts on serial sections indicated that 11.5% of adult male CBA thymic lymphoid cells were within the medullary zone. Since only 3-4% of thymocytes were cortisone resistant, the majority of thymocytes within the medulla were, like cortical thymocytes, cortisone sensitive. A series of cell surface antigenic markers, used alone or in pairs, suggested that 13-15% of thymocytes were of medullary phenotype, somewhat more than the number of thymocytes actually present in the medulla. However, much of this discrepancy could be explained by differential death of cortical cells during isolation and staining, and by the existence in the cortex of a subpopulation of early blast cells which shared some, but not all markers with medullary thymocytes. A direct test for mature or medullary phenotype cells in the cortex involved selective transcapsular labeling of outer-cortical cells with fluorescent dyes, followed by multiparameter immunofluorescent analysis of the 10% labeled population. Outer-cortical thymocytes included some cells (mainly early blasts) sharing some markers with medullary thymocytes, but very few (less than 1%) of these cells expressed all the characteristic "mature" markers. Limit-dilution precursor frequency studies showed the level of functional cells in the outer cortex was extremely low. The overall conclusion was that the vast majority of cells of complete "mature" phenotype are confined to the thymic medulla. These findings favor the view that thymus migrants originate from the thymic medulla, but do not exclude a cortical origin. The results also illustrate the need for multiparameter analysis to distinguish medullary thymocytes from early blast cells.     

The effect of historical legacy on adaptation: do closely related species respond to the environment in the same way?

The many documented examples of parallel and convergent evolution in similar environments are strong evidence for the role of natural selection in the evolution of trait variation. However, species may respond to selection in different ways; idiosyncrasies of their evolutionary history may affect how different species respond to the same selective pressure. To determine whether evolutionary history affects trait-environment associations in a recently diverged lineage, we investigated within-species trait-environment associations in the white proteas, a closely related monophyletic group. We first used manovas to determine the relative importance of shared response to selection, evolutionary history and unique responses to selection on trait variation. We found that on average, similar associations to the environment across species explained trait variation, but that the species had different mean trait values. We also detected species-specific associations of traits with the environmental gradients. To identify the traits associated uniquely with the environment, we used a structural equation model. Our analysis showed that the species differed in how their traits were associated with each of the environmental variables. Further, in the cases of two root traits (root mass and root length/mass ratio), two species differed in the direction of their associations (e.g. populations in one species had heavier roots in warmer areas, and populations in the other species had lighter roots in warmer areas). Our study shows that even in a closely related group of species, evolutionary history may have an effect on both the size and direction of adaptations to the environment.     

The NO way to increase muscular utrophin expression?

Duchenne muscular dystrophy (DMD), a severe X-linked recessive disorder that results in progressive muscle degeneration, is due to a lack of dystrophin, a membrane cytoskeletal protein. An approach to the search for a treatment is to compensate for dystrophin loss by utrophin, another cytoskeletal protein. During development, in normal as in dystrophic embryos, utrophin is found at the membrane surface of immature skeletal fibres and is progressively replaced by dystrophin. Thus, it is possible to consider utrophin as a 'foetal homologue' of dystrophin. In a previous work, we studied the effect of L-arginine, the substrate of nitric oxide synthetase (NOS), on utrophin expression at the muscle membrane. Using a novel antibody, we confirm here that the immunocytochemical staining was indeed due to an increase in utrophin at the sarcolemma. The result is observed not only on mdx (an animal model of DMD) myotubes in culture but also in mdx mice treated with L-arginine. In addition, we show here the utrophin increase in muscle extracts of mdx mice treated with L-arginine, after electrophoretic separation and western-blotting using this novel antibody, and thus extending the electrophoretic results previously obtained on myotube cultures to muscles of treated mice.     

Promiscuous tendencies in the round-eared sengi: a male’s perspective

Although monogamy is interpreted as risk-adverse strategy by reducing intra-sexual conflicts, most pair-living males increase their reproductive success by engaging in extra-pair copulations. However, little is known about costs involved in such extra-pair attempts from the male’s perspective. We investigated promiscuous tendencies of paired Macroscelides proboscideus (round-eared elephant-shrew or round-eared sengi), a pair-living small mammal occurring in southern Africa. In particular, we measured potential costs of extra-pair attempts for the males. For this, we conducted laboratory experiments, involving interactions between neighbouring male–female pairs. Data collection included direct behavioural observations and establishing the morphological and physiological characteristics of males. Both sexes intruded into the neighbouring area, and initiated sexual behaviour with neighbours of the opposite sex. Males which displayed a higher marking frequency in the neighbouring area received more sexually motivated behaviour from neighbouring females. Resident males attacked intruding males. Aggression experienced by and marking behaviour of intruding males was positively correlated with body mass losses. Furthermore, glucocorticoid levels, determined from analyses of faeces and urine samples, correlated positively with body mass losses and marking behaviour of intruding males, indicating costs of intruding. We conclude that male as well as female round-eared sengis have promiscuous tendencies. Although interactions with neighbouring females may offer potential benefits for males in terms of enhanced reproductive success, males also seem to pay substantial costs, as indicated by body mass losses and elevated stress hormone levels.     

ARF in the mitochondria: The last frontier?

The tumor suppressor ARF carries out different functions in different cellular compartments. In the nucleus, ARF interacts physically and functionally with Mdm2 to inhibit cell cycle progression through activation of p53. In the nucleolus, ARF interacts with B23/NPM to inhibit ribosomal biogenesis through control of rRNA processing. Recent studies have expanded ARF's territory into the mitochondria. New data have shown that ARF interacts with the mitochondrial protein p32/C1QBP and that the interaction is critical in order for ARF to localize to the mitochondria and induce apoptosis. Remarkably, the ARF-p32 interaction, and hence ARF's pro-apoptotic function, can be interrupted by human cancer-derived mutations in exon2 of the p14ARF-p16INK4a gene locus. Here, we discuss the implications of these studies and their potential relevance to human cancer.     

The glutamine commute: Lost in the tube?

The "glutamate-glutamine" cycle appears to have an important, albeit not exclusive role, in the recycling of glutamate (Glu) between neurons and astrocytes. Recent studies show that the efflux of glutamine (Gln) from astrocytes is mediated by SNAT3 (formerly SN1), a system N amino acid transporter localized to perisynaptic astrocytes, whereas its influx into neurons is thought to be mediated by transporters of the system A family, specifically SNAT1 and SNAT2. However, the results of our confocal and electron microscopy immunocytochemical studies of the localization of these transporters in the cerebral cortex show that SNAT1 and SNAT2 are robustly expressed in the somatodendritic domain of cortical neurons, but rarely to axon terminals. To rule out a possible influence of fixation and procedural variables on detection of SNAT1 and SNAT2 immunoreactivity in axon terminals, we used non-conventional immunocytochemical methods, which, in certain cases, improve antigen detection. Though evidencing a slightly increased percentage of axon terminals expressing the two transporters, these techniques demonstrated that SNAT1 and SNAT2 are indeed rarely localized to axon terminals. Our data thus suggest that neither SNAT1 nor SNAT2 meet the criteria for their postulated role in the "glutamate-glutamine" cycle, and indicate that other Gln transporters (either orphan or yet to be identified) must be expressed at axon terminals and sustain the Glu (and gamma-aminobutyric acid) neurotransmitter pool (s).     

The CBM signalosome: Potential therapeutic target for aggressive lymphoma?

The CBM signalosome plays a pivotal role in mediating antigen-receptor induced NF-κB signaling to regulate lymphocyte functions. The CBM complex forms filamentous structure and recruits downstream signaling components to activate NF-κB. MALT1, the protease component in the CBM complex, cleaves key proteins in the feedback loop of the NF-κB signaling pathway and enhances NF-κB activation. The aberrant activity of the CBM complex has been linked to aggressive lymphoma. Recent years have witnessed dramatic progresses in understanding the assembly mechanism of the CBM complex, and advances in the development of targeted therapy for aggressive lymphoma. Here, we will highlight these progresses and give an outlook on the potential translation of this knowledge from bench to bedside for aggressive lymphoma patients.     

Low expression of Neu2 sialidase in the thymus of SM/J mice—existence of neuraminidase positive cells “Neu-medullocyte” in the murine thymus

We have already reported that the homogenate of the A/J mouse thymus shows a high sialidase activity at the neutral pH region and that in both soluble and membrane fractions optimal pH was 6.5–7 (Kijimoto-Ochiai et al., Glycoconj. J., 20:375–384, 2004). In the present study, we investigated the level of sialidase activities in the thymus of the SM/J mouse, a mouse strain that we know to have a Neu1^a allele that reveals a low level of sialidase activity in the liver. We found that while in the A/J thymus the soluble sialidase activity at pH 6.5 was high, the SM/J thymus lacked all such activity. A QTL analysis of SMXA recombinant inbred strains showed that soluble sialidase activity correlated well with the D1Mit8/9 marker on chromosome 1. The murine whole DNA-sequence data and the results of our FISH analysis (Kotani et al., Biochem. Biophys. Res. Comm., 286:250–258, 2001) showed that this location is consistent with the position of Neu2 gene. We confirmed that it is hard to detect the Neu2 enzyme of the SM/J mouse thymus by an anti-Neu2 antibody using a Western blot analysis. We also found that while the mRNA expression of Neu2 was quite normal in the SM/J mouse liver, it was very low in the SM/J mouse thymus. We therefore conclude that the lack of soluble sialidase activity in the SM/J mouse thymus is due to the thymus-specific low expression level of the Neu2 gene. We have previously shown that the sialidase positive cell which contains the Mac-1 and immunoglobulin, and which is located sparsely in the corticomedullar region or medullary region of the A/J mouse thymus (Kijimoto-Ochiai et al., Glycoconj. J., 20:375–384, 2004). We showed now in this paper that the detection of this cell in the SM/J mouse thymus at pH 7.0 was difficult. We propose, therefore, to name the cell “Neu-medullocyte”.