RIC-3 expression and splicing regulate nAChR functional expression.

BACKGROUND: The nicotinic acetylcholine receptors form a large and diverse family of acetylcholine gated ion channels having diverse roles in the central nervous system. Maturation of nicotinic acetylcholine receptors is a complex and inefficient process requiring assistance from multiple cellular factors including RIC-3, a functionally conserved endoplasmic reticulum-resident protein and nicotinic acetylcholine receptor-specific chaperone. In mammals and in Drosophila melanogaster RIC-3 is alternatively spliced to produce multiple isoforms. RESULTS: We used electrophysiological analysis in Xenopus laevis oocytes, in situ hybridization, and quantitative real-time polymerase chain reaction assays to investigate regulation of RIC-3's expression and splicing and its effects on the expression of three major neuronal nicotinic acetylcholine receptors. We found that RIC-3 expression level and splicing affect nicotinic acetylcholine receptor functional expression and that two conserved RIC-3 isoforms express in the brain differentially. Moreover, in immune cells RIC-3 expression and splicing are regulated by inflammatory signals. CONCLUSIONS: Regulation of expression level and splicing of RIC-3 in brain and in immune cells following inflammation enables regulation of nicotinic acetylcholine receptor functional expression. Specifically, in immune cells such regulation via effects on α7 nicotinic acetylcholine receptor, known to function in the cholinergic anti-inflammatory pathway, may have a role in neuroinflammatory diseases.

Regulation of osteoclastogenesis by integrated signals from toll-like receptors.

A variety of pathogen-derived molecules have been shown to cause bone loss by enhancing osteoclast differentiation through activation of toll-like receptors (TLRs). The pathogen-derived molecules (TLR-ligands) modulate osteoclastogenesis in a complex manner: inhibition of the osteoclast differentiation factor RANKL in early precursors and osteoclastogenesis stimulation in RANKL-primed cells. Since organisms may be challenged by several TLR ligands at a time, we investigated osteoclastogenesis modulation by simultaneous challenge with different TLR ligands. As an example we used ligands for TLR3 (Synthetic double stranded RNA [dsRNA], polyinosinic-polycytidylic acid [poly(I:C)] mimicking viral dsRNA), TLR4 (lipopolysaccharide [LPS], found in the outer membrane of Gram-negative bacteria) and TLR9 (Synthetic oligodeoxynucleotide mimicking bacterial DNA [CpG-ODN]). In osteoclastogenesis-inhibition, synergy between LPS and CpG-ODN or LPS and poly(I:C) while in stimulation, synergy between LPS and CpG-ODN or CpG-ODN and poly(I:C) were observed. Modulation of molecules involved in osteoclastogenesis (c-Fos, M-CSF receptors [M-CSFR], TNF-α, IL-6, and IL-12 and the three TLRs tested) was examined. The results indicate that M-CSFR plays a role only in the inhibitory effect while c-Fos plays a role in the two effects. TLR3 and TLR9 levels were increased by the TLRs ligands, suggesting that this may be part of the mechanism leading to the synergy. While TLRs activation in RANKL-primed cells, increasing osteoclastogenesis, explains pathogen-induced bone loss, activation of TLRs in early cells inhibiting osteoclastogenesis could attenuate excessive resorption, and promote differentiation of common precursor cells into inflammatory cells. The synergism between TLR ligands enables the individual to initiate response at a lower level of pathogen.