Lipopolysaccharide (LPS) preconditioning reduces ischemic injury in adult brain by activating Toll-like receptor 4 (TLR-4). We sought to investigate the effect of brain maturity on the efficacy of LPS preconditioning against hypoxic-ischemic (HI) injury in the developing rat brain. Rat pups at the specified age were randomly assigned to LPS-treated (0.1 mg/kg) or saline-treated groups. HI injury was induced 48 h later by occluding the right common carotid artery followed by transient hypoxia. Brains were removed 1 wk after HI injury, and infarct volumes were compared between the two groups. TLR-4 expression was also compared among different ages. We found that LPS treated P7, P9, and P14 rat pups had significantly smaller infarct volume compared with saline-treated pups (p ϭ 0.006, 0.03, and 0.01, respectively). This significant reduction in infarct volume was not observed in P3 and P5 rats. TLR-4 expression was significantly higher in older rats compared with P3 and P5 rats (p Ͻ 0.01). These findings indicate that LPS-induced preconditioning is a robust neuroprotective phenomenon in the ischemic developing brain that is age dependent. Pattern of TLR-4 expression is also affected by brain maturity and likely to be responsible for differences in the efficacy of LPS preconditioning. (Pediatr Res 70: 10-14, 2011) D elayed preconditioning is a potent protective phenomenon in which a tissue like the brain develops resistance to ischemic injury after exposure to a variety of subinjurious stimuli such as brief ischemia, hypoxia, or low dose of lipopolysaccharide (LPS) (1-5). This neuroprotective effect is prolonged and can last for several days after the exposure to the subinjurious stimulus. LPS neuroprotection is well documented in adult stroke animal models (6 -9), whereas studies on LPS preconditioning in the hypoxic-ischemic (HI) immature brain are scarce (10,11). In contrast to ischemic adult brain, LPS given to postnatal d 7 (P7) rat pups 72 h before HI injury increased brain damage but was neuroprotective when administered 24 h before the HI insult (10). Recently, we have shown that LPS preconditioning protected against cardiopulmonary bypass-related brain injury in neonatal piglets when administered 72 h before the procedure (11). These conflicting results on the effect of LPS treatment before HI injury in the developing brain may be attributed to differences in LPS doses, time interval between LPS treatment and the severity of the HI insult. Moreover, the influence of brain maturity on the pathophysiology of ischemic injury is well documented (12)(13)(14). It is conceivable, therefore, that there may be a critical developmental window at which the phenomenon of delayed preconditioning can be used to protect the developing brain against ischemic injury.LPS is a known specific ligand for Toll-like receptor 4 (TLR-4), one of the Toll-like receptors (TLRs) that recognize foreign pathogens. LPS-mediated ischemic tolerance in the adult brain occurs by stimulation of TLR-4 signaling pathways (6,15), result...