SUMMARY The molecular determinants of spleen organogenesis and the etiology of isolated congenital asplenia (ICA), a life-threatening human condition, are unknown. We previously reported that Pbx1 deficiency causes organ growth defects including asplenia. Here, we show that mice with splenic mesenchyme-specific Pbx1 inactivation exhibit hyposplenia. Moreover, the loss of Pbx causes down-regulation of Nkx2-5 and derepression of p15Ink4b in spleen mesenchymal progenitors, perturbing the cell cycle. Removal of p15Ink4b in Pbx1 spleen-specific mutants partially rescues spleen growth. By whole-exome sequencing of a multiplex kindred with ICA, we identify a heterozygous missense mutation (P236H) in NKX2-5 showing reduced transactivation in vitro. This study establishes that a Pbx/Nkx2-5/p15 regulatory module is essential for spleen development.

Dendritic growth is the common mode of solidification encountered when metals and alloys freeze under low thermal gradients. The growth of dendrites in pure melts depends on the transport of latent heat from the moving crystal-melt interface and the influence of weaker effects like the interfacial energy. Experimental data for critical tests of dendritic growth theories remained limited because dendritic growth can be complicated by convection. The Isothermal Dendritic Growth Experiment (IDGE) was developed specifically to test dendritic growth theories by performing measurements with succinonitrile (SCN) in microgravity, thus eliminating buoyancy-induced convection. The first flight of the IDGE in 1994 operated for 9 days at a mean quasi-static acceleration of 0.7 ϫ 10 Ϫ6 g 0 . The velocity and radius data show that at supercoolings above approximately 0.4 K, dendritic growth in SCN under microgravity conditions is diffusion limited. By contrast, under terrestrial conditions, dendritic growth of SCN is dominated by convection for supercoolings below 1.7 K. The theoretical and experimental Péclet numbers exhibit modest disagreement, indicating that transport theories of dendritic solidification require some modification. Finally, the kinetic selection rule for dendritic growth, VR 2 ϭ constant, where V is the velocity of the tip and R is the radius of curvature at the tip, appears to be independent of the gravity environment, with a slight dependence on the supercooling.

We measured dendritic tip velocities in pure succinonitrile (SCN) in microgravity, using a sequence of telemetered binary images sent to Earth from the space shuttle Columbia (STS-62). Growth velocities were measured as a function of the supercooling over the range 0.05 -1.5 K. Microgravity observations show that buoyancy-induced convection alters the growth kinetics of SCN dendrites at supercoolings as high as 1.3 K. Also, the dendrite velocity data measured under microgravity agree well with the Ivantsov paraboloidal diffusion solution when coupled to a scaling constant of o. * = 0.0157.

TALE-homeodomain proteins function as components of heteromeric complexes that contain one member each of the PBC and MEIS/ PREP subclasses. We recently showed that MEIS2 cooperates with the neurogenic transcription factor PAX6 in the control of adult subventricular zone (SVZ) neurogenesis in rodents. Expression of the PBC protein PBX1 in the SVZ has been reported, but its functional role(s) has not been investigated. Using a genetic loss-of-function mouse model, we now show that Pbx1 is an early regulator of SVZ neurogenesis. Targeted deletion of Pbx1 by retroviral transduction of Cre recombinase into Pbx2-deficient SVZ stem and progenitor cells carrying floxed alleles of Pbx1 significantly reduced the production of neurons and increased the generation of oligodendrocytes. Loss of Pbx1 expression in neuronally committed neuroblasts in the rostral migratory stream in a Pbx2 null background, by contrast, severely compromised cell survival. By chromatin immunoprecipitation from endogenous tissues or isolated cells, we further detected PBX1 binding to known regulatory regions of the neuron-specific genes Dcx and Th days or even weeks before the respective genes are expressed during the normal program of SVZ neurogenesis, suggesting that PBX1 might act as a priming factor to mark these genes for subsequent activation. Collectively, our results establish that PBX1 regulates adult neural cell fate determination in a manner beyond that of its heterodimerization partner MEIS2.