Know thy neighbor: Modeling spatiotemporal cell-fate patterns in a neural stem cell niche.

How lineage and the microenvironment influence stem cell homeostasis at a population level remains unresolved. In this issue of Cell Stem Cell, Dray et al. (2021) use in vivo imaging and statistical modeling to discover a key role for local progenitor cell descendants in constraining neural stem cell divisions.

Treatment of neutral glycosphingolipid lysosomal storage diseases via inhibition of the ABC drug transporter, MDR1. Cyclosporin A can lower serum and liver globotriaosyl ceramide levels in the Fabry mouse model.

We have shown that the ABC transporter, multiple drug resistance protein 1 (MDR1, P-glycoprotein) translocates glucosyl ceramide from the cytosolic to the luminal Golgi surface for neutral, but not acidic, glycosphingolipid (GSL) synthesis. Here we show that the MDR1 inhibitor, cyclosporin A (CsA) can deplete Gaucher lymphoid cell lines of accumulated glucosyl ceramide and Fabry cell lines of globotriaosyl ceramide (Gb3), by preventing de novo synthesis. In the Fabry mouse model, Gb3 is increased in the heart, liver, spleen, brain and kidney. The lack of renal glomerular Gb3 is retained, but the number of verotoxin 1 (VT1)-staining renal tubules, and VT1 tubular targeting in vivo, is markedly increased in Fabry mice. Adult Fabry mice were treated with alpha-galactosidase (enzyme-replacement therapy, ERT) to eliminate serum Gb3 and lower Gb3 levels in some tissues. Serum Gb3 was monitored using a VT1 ELISA during a post-ERT recovery phase +/- biweekly intra peritoneal CsA. After 9 weeks, tissue Gb3 content and localization were determined using VT1/TLC overlay and histochemistry. Serum Gb3 recovered to lower levels after CsA treatment. Gb3 was undetected in wild-type liver, and the levels of Gb3 (but not gangliosides) in Fabry mouse liver were significantly depleted by CsA treatment. VT1 liver histochemistry showed Gb3 accumulated in Kupffer cells, endothelial cell subsets within the central and portal vein and within the portal triad. Hepatic venule endothelial and Kupffer cell VT1 staining was considerably reduced by in vivo CsA treatment. We conclude that MDR1 inhibition warrants consideration as a novel adjunct treatment for neutral GSL storage diseases.

Reduced trabecular bone mass and strength in mice overexpressing Gα11 protein in cells of the osteoblast lineage.

G protein-coupled receptors (GPCRs) require G proteins for intracellular signaling to regulate a variety of growth and maintenance processes, including osteogenesis and bone turnover. Bone maintenance events may be altered by changes in the activity or level of G proteins, which then modify signaling in bone cells such as osteoblasts. We have previously reported increased levels of Gα11 protein and signaling to phospholipase C/protein kinase C pathways in response to dexamethasone in osteoblastic UMR 106-01 cells. Here we generated pOBCol3.6-GNA11 transgenic mice that overexpress Gα11 protein in cells of the osteoblast lineage (G11-Tg mice). G11-Tg mice exhibit an osteopenic phenotype characterized by significant reductions in trabecular bone mineral density, thickness, number and strength. The numbers of osteoblasts and osteocytes were unchanged in G11-Tg bone, but early markers of osteoblast differentiation, Alp and Bsp, were increased while the late stage differentiation marker Ocn was not changed suggesting reduced osteoblast maturation in G11-Tg trabecular bone which was accompanied by a decreased bone formation rate. Furthermore, in vitro cultures of G11-Tg primary osteoblasts show delayed osteoblast differentiation and mineralization. Histological analyses also revealed increased osteoclast parameters, accompanied by elevated mRNA expression of Trap and Ctsk. mRNA levels of Rankl and M-csf were elevated in vitro in bone marrow stromal cells undergoing osteogenesis and in trabecular bone in vivo. Together, these findings demonstrate that increasing Gα11 protein expression in osteoblasts can alter gene expression and result in a dual mechanism of trabecular bone loss.

Waste management and adult neurogenesis.

Dying cells are common in adult neurogenic niches, but how these cells are cleared remains uncertain. In this issue of Cell Stem Cell, Sierra et al. (2010) show that unactivated microglia assume the role of waste managers to eliminate cellular debris from apoptosing newborn cells in the hippocampus.

Regulation of D1 dopamine receptor trafficking and signaling by caveolin-1.

There is accumulating evidence that G protein-coupled receptor signaling is regulated by localization in lipid raft microdomains. In this report, we determined that the D1 dopamine receptor (D1R) is localized in caveolae, a subset of lipid rafts, by sucrose gradient fractionation and confocal microscopy. Through coimmunoprecipitation and bioluminescence resonance energy transfer assays, we demonstrated that this localization was mediated by an interaction between caveolin-1 and D1R in COS-7 cells and an isoform-selective interaction between D1R and caveolin-1alpha in rat brain. We determined that the D1R interaction with caveolin-1 required a putative caveolin binding motif identified in transmembrane domain 7. Agonist stimulation of D1R caused translocation of D1R into caveolin-1-enriched sucrose fractions, which was determined to be a result of D1R endocytosis through caveolae. This was found to be protein kinase A-independent and a kinetically slower process than clathrin-mediated endocytosis. Site-directed mutagenesis of the caveolin binding motif at amino acids Phe313 and Trp318 significantly attenuated caveolar endocytosis of D1R. We also found that these caveolin binding mutants had a diminished capacity to stimulate cAMP production, which was determined to be due to constitutive desensitization of these receptors. In contrast, we found that D1Rs had an enhanced ability to maximally generate cAMP in chemically induced caveolae-disrupted cells. Taken together, these data suggest that caveolae has an important role in regulating D1R turnover and signaling in brain.