Propylene glycol releases calcium from mitochondrial stores in rat cerebrocortical synaptosomes

In Ca2+-free medium, propylene glycol (PG) increases the cytosolic-free Ca2+ concentration ([Ca2+]i) in rat cerebrocortical synaptosomes. In the current studies, the authors investigated the intracellular source of this Ca2+. PG (0.5-5% v/v) dose-dependently increased [Ca2+]i in Ca2+-free medium. The increase in [Ca2+]i was completely inhibited by pretreatment with the mitochondrial uncoupler, carbonylcyanide m-chlorophenylhydrazone (CCCP, 10 microM). The inhibitory effect of CCCP was dependent on the concentration and the pretreatment time. These results suggest that, in Ca2+-free medium, PG increases [Ca2+]i in rat cerebrocortical synaptosomes by releasing Ca2+ from mitochondrial stores.     

Distinct migratory behavior of early- and late-born neurons derived from the cortical ventricular zone

Time-lapse studies indicate that ventricular zone (VZ)-derived cells show two migratory modes in the cerebral cortex at different stages of mammalian embryogenesis: somal translocation and locomotion. We carried out a systematic analysis to examine whether the migratory behavior of cortical neurons derived from the cortical VZ is stage-dependent. We labeled VZ cells of mouse embryos with green fluorescent protein (gfp) -encoding plasmids by in utero electroporation and evaluated the labeled cells after appropriate survival periods. After electroporation at either embryonic day (E) 12.5 or E15.5, GFP+ VZ cells were initially spindle-shaped and radially oriented. After leaving the VZ, they transformed into round or horizontally oriented fusiform neurons with many short processes. They then seemed to gradually change into radially oriented bipolar cells as they moved upward. Whereas the earliest emigrants from the VZ labeled at E12.5 (early-born neurons) reached the top of the cortical plate (CP) after these changes, VZ cells labeled at E15.5 (late-born neurons) further migrated along the length of radial fibers to reach the top of the CP. A dominant negative form of the gene for cyclin-dependent kinase 5 (Cdk5DN) was then introduced into VZ cells. Transfection of E12.5 VZ with cdk5dn did not disrupt the migration of the early-born neurons. However, this caused a failure in migration of the late-born neurons, although they transformed into bipolar shapes in the intermediate zone. Thus, there appear to be at least two distinct migratory phases of cortical neurons: one common to the early- and late-born neurons, and the other specific to late-born neurons and Cdk5-dependent.     

HTLV-I carrier with unusual brain MR imaging findings

We describe unusual brain MR imaging findings in a patient who is an HTLV-I carrier without myelopathy. T2-weighted MR images showed hyperintense signal abnormalities in the pyramidal tract, superior and middle cerebellar peduncles, and decussation of the superior cerebellar peduncles, in addition to subcortical white matter involvement. Diffusion-weighted images also showed hyperintense signal abnormalities in the same regions by T2 shine-through effect.     

Ontogenic expression of estrogen receptor coactivators in the reproductive tract of female mice neonatally exposed to diethylstilbestrol

Ontogenic expression of p300, steroid receptor coactivator-1 (SRC-1), GR-interacting protein-1 (GRIP1) and p300/CBP cointegrator associate protein (p/CIP) was examined using an immunohistochemical method in the genital tract of female mice neonatally exposed to diethylstilbestrol (DES). Mice were injected with 4 microg of DES for 5 days and killed on days 10, 15 and 21; some mice were killed on day 4 after three injections. The p300 immunoreactive protein in the epithelial cells of the oviducts, uteri, and vaginae was almost constant during development independent of neonatal DES exposure, while the stromal cells exhibited a weaker reaction than that of the vehicle-treated controls on days 4 and 10. Neonatal DES exposure caused no significant changes in the SRC-1 and p/CIP expression patterns but decreased the GRIP1 expression with development in the reproductive tract. The constitutive expression of coactivators in both epithelial and stromal cells may play a role in the estrogen imprints through ER alpha systems during the period of DES treatment.     

Association of N-myc downregulated gene 1 with heat-shock cognate protein 70 in mast cells

N-Myc downregulated gene (NDRG) 1 is markedly induced during in vitro maturation of mouse immature bone marrow-derived mast cells (BMMCs) into a mature connective tissue mast cell (CTMC)-like phenotype. However, cellular function of this unique cytosolic protein is currently obscure. In this study, we sought potential NDRG1-binding proteins using yeast two-hybrid analysis and found that NDRG1 is capable of binding to heat-shock cognate protein 70 (Hsc70) both in vitro and in mast cells. The expression of Hsc70 was markedly elevated during the in vitro maturation of BMMCs into CTMC-like cells in accordance with the increased expression of NDRG1. Deletion of the C-terminal hydrophilic tandem repeats from NDRG1 facilitated the interaction with Hsc70 in vitro. Interaction between NDRG1 and Hsc70 was constitutive in mast cells and was not altered following cell activation. Although NDRG1 undergoes phosphorylation (accompanying paper), the binding of NDRG1 to Hsc70 was not affected by this event. Interestingly, the NDRG1-Hsc70 complex transiently appeared in the nuclear fraction of activated mast cells.     

Clinicopathological study on suprachoroidal and supraciliary hemorrhage during enucleation

PURPOSE: To investigate the pathogenesis of suprachoroidal and supraciliary hemorrhage that might have been induced during enucleation. METHODS: A histopathological examination of 392 enucleated eyeballs was carried out and 8 eyeballs with suprachoroidal and supraciliary hemorrhage were selected for further clinicopathological examination. RESULTS: Among 14 eyeballs with severe acute intraocular inflammation, 7 eyeballs with suprachoroidal and supraciliary hemorrhage were found and one other such eyeball was seen among 53 with neovascular glaucoma. Among these 8, there was one case of prolapse of intraocular tissue with severe hemorrhage into the suprachoroidal and supraciliary spaces; 4 cases of severe hemorrhage into the suprachoroidal and supraciliary spaces without prolapse of intraocular tissue; and 3 cases of mild hemorrhage into the suprachoroidal or supraciliary spaces. CONCLUSIONS: In the eyeballs with severe acute intraocular inflammation, intraocular pressure was elevated and the blood vessels were weakened by inflammatory cell infiltration. During enucleation external forces affected the blood vessel wall of the ciliary arteries and vortex veins, and the breakdown of vessel walls might have been the cause of the suprachoroidal and supraciliary hemorrhage.     

Diquafosol elicits increases in net Cl- transport through P2Y2 receptor stimulation in rabbit conjunctiva

The purpose of the present study was to understand the mechanisms of action of diquafosol, a stable derivative of uridine 5'-triphosphate, on Cl(-) transport across the isolated rabbit conjunctiva. Rabbit conjunctivas were isolated and mounted in a modified Ussing chamber. Under short-circuit conditions, the effects were determined of mucosal (tear) side diquafosol application on the short-circuit current (Isc). Diquafosol rapidly and dose-dependently increased the Isc at concentrations ranging from 0.1 to 968 microM when added to the mucosal side of the conjunctiva. In the absence of the serosal Cl(-), the Isc induced by 10 microM diquafosol was substantially reduced. On the contrary, in the absence of mucosal side Na(+), the diquafosol-induced increases in Isc were unchanged. Following 45-min preincubation, the P2Y(2) antagonist suramin inhibited the diquafosol-induced increases in the Isc whereas the P2Y(1) antagonist pyridoxal-phosphate-6-azophenyl-2'4'-disulfonic acid had no effect. These studies suggest that diquafosol stimulates net Cl(-) secretion from the serosal to the mucosal side via stimulation of P2Y(2) receptors in the rabbit conjunctiva.