We used single-cell reverse transcription and polymerase chain reaction (RT-PCR) to determine whether dorsomedial pontine cells with projections to the mMRF express mRNA for selected membrane receptors that mediate modulatory influences on REM sleep. Fluorescein (FITC)-labeled latex microspheres were microinjected into the mMRF of 26-34-day-old rats under pentobarbital anesthesia. After 5-6 days, rats were sacrificed, pontine slices were obtained and neurons were dissociated from 400 to 600 mu m micropunches extracted from dorsomedial pontine

reticular formation. We found that 32 out of 51 FITC-labeled cells tested (63 PRT062607 clinical trial +/- 7% (SE)) contained the orexin type I receptor (ORX1r) mRNA, 27 out of 73 (37 +/- 6%) contained the adrenergic alpha(2A) receptor (alpha(2A)r) RNA, and 6 out of 31 (19 7%) contained both mRNAs. The percentage of cells positive for the ORX1r mRNA was significantly lower (p < 0.04) for the dorsomedial pontine cells that were not retrogradely labeled from the mMRF (32 +/- 11 %), whereas alpha(2A)r

mRNA was present in a similar percentage of FITC-labeled and unlabeled neurons. Our data suggest that ORX and adrenergic pathways converge on a subpopulation of cells of the pontine REM sleep-triggering region that have descending this website projections to the medullary region important for the motor control during REM sleep. (C) 2009 Elsevier Ireland Ltd. All rights reserved.”
“Epstein-Barr virus (EBV) infection is mediated by several viral envelope glycoproteins. We have assessed gp110′s functions during the virus life cycle using a mutant that lacks BALF4 (Delta BALF4). Exposure of various cell lines and primary cell samples of epithelial or lymphoid lineages to the Delta BALF4 mutant failed to establish stable infections. The Delta BALF4 virus, however, did not differ from wild-type EBV in its ability to bind and become internalized into primary B cells, in which it elicited a potent T-cell-specific immune reaction against virion constituents. These findings show that Delta BALF4 viruses can reach the endosome-lysosome compartment and dovetail nicely with

the previously identified contribution of gp110 to virus-cell fusion. Other essential steps of the Selleck Dibutyryl-cAMP virus life cycle were unaffected in the viral mutant; DNA lytic replication and viral titers were not altered in the absence of gp110, and Delta BALF4 viruses complemented in trans transformed infected B cells with an efficiency indistinguishable from that observed with wild-type viruses. All of the steps of virus maturation could be observed in lytically induced 293/Delta BALF4 cells. Induction of lymphoblastoid cells generated with transiently complemented Delta BALF4 virus led to the production of rare mature virions. We therefore infer that gp110 is not required for virus maturation and egress in 293 cells or in B cells.