Anti-NLRP2 (full length) polyclonal antibody

Purpose The age-associated decline in female fertility is largely ascribable to the decrease in oocyte quality. The subcortical maternal complex (SCMC) is a multiprotein complex essential for early embryogenesis and female fertility and functionally conserved across mammals. The present work evaluated expression dynamics of its components during folliculogenesis in relation to maternal age in sheep. Methods The expression of the SCMC components (KHDC3/FILIA,NLRP2,NLRP5/MATER,OOEP/FLOPED,PADI6,TLE6andZBED3) was analyzed by real-time PCR in pools of growing oocytes (GO) of different diameters (70-90 mu m (S), 90-110 mu m (M), or 110-130 mu m (L)) derived from non-hormonally treated adult (Ad; age < 4 years), prepubertal (Pr; age 40 days), or aged ewes (age > 6 years). Results Specific expression patterns associated with donor age were observed during folliculogenesis for all genes, exceptZBED3. In oocytes of adult donors, the synthesis ofNLRP2,NLRP5,PADI6, andZBED3mRNAs was complete in S GO, whileFILIA,TLE6, andOOEPwere actively transcribed at this stage. Conversely, Pr GO showed active transcription of all mRNAs, except forZBED3, during the entire window of oocyte growth. Notably, aged GO showed a completely inverse pattern, with a decrease ofNLRP2,TLE6,FILIA, andPADI6mRNA abundance during the latest stage of oocyte growth (L GO). Interestingly,MATERshowed high expression variability, suggesting large inter-oocyte differences. Conclusion Our study describes the SCMC expression dynamics during sheep oogenesis and reports age-specific patterns that are likely involved in the age-related decline of oocyte quality.

Background Systemic hypertension is a risk factor of age-related macular degeneration (AMD), a chronic inflammatory disease. Acute hypertension is caused by increased extracellular osmolarity after intake of dietary salt (NaCl). We determined in cultured human retinal pigment epithelial (RPE) cells whether high extracellular NaCl alters the gene expression of inflammasome-associated proteins, and whether autocrine/paracrine purinergic (P2) receptor signaling contributes to the NaCI-induced NLRP3 gene expression. Methodology/Principal Findings Hyperosmolarity was induced by the addition of 100 mM NaCl or sucrose to the culture medium. Gene and protein expression levels were determined with real-time RT-PCR and Western blot analysis, respectively. IL-1 beta and IL-18 levels were evaluated with ELISA. Nuclear factor of activated T cell 5 (NFAT5) expression was knocked down with siRNA. High extracellular NaCI induced NLRP3 and pro-IL-1 beta gene expression, while the gene expression of further inflammasome-associated proteins (NLRP1, NLRP2, NLRP6, NLRP7, NLRP12, NLRC4, AIM2, ASC, procaspase-1, pro-IL-18) was not altered or below the detection threshold. The NaCI-induced NLRP3 gene expression was partially dependent on the activities of phospholipase C, IP3 receptors, protein kinase C, the serum and glucocorticoid-regulated kinase, p38 MAPK, ERK1/2, JNK, P13K, and the transcription factors HIF-1 and NFAT5. Pannexin-dependent ATP release and P2Y(1) receptor activation is required for the full induction of NLRP3 gene expression. High NaCI induced a transient increase of the NLRP3 protein level and a moderate NLRP3 inflammasome activation, as indicated by the transient increase of the cytosolic level of mature IL-1 beta. High NaCl also induced secretion of IL-18. Conclusion High extracellular NaCl induces priming of the NLRP3 inflammasome in RPE cells, in part via P2Y(1) receptor signaling. The inflammasome priming effect of NaCl suggests that high intake of dietary salt may promote local retinal inflammation implicated in the development of AMD.