Anti-LRP8 (aa 83-170) polyclonal antibody

Coordinated migration of neurons in the developing and adult brain is essential for its proper function. The secreted glycoprotein Reelin (also known as RELN) guides migration of neurons by binding to two lipoprotein receptors, the very-low-density lipoprotein receptor (VLDLR) and apolipoprotein E receptor 2 (ApoER2, also known as LRP8)(1). Loss of Reelin function in humans results in the severe developmental disorder lissencephaly(2) and it has also been associated with other neurological disorders such as epilepsy, schizophrenia and Alzheimer's disease(3). The molecular mechanisms by which Reelin activates its receptors and controls cellular functions are largely unknown. Here we show that the neuronal guidance cues ephrin B proteins are essential for Reelin signalling during the development of laminated structures in the brain. We show that ephrin Bs genetically interact with Reelin. Notably, compound mouse mutants (Reln(+/-); Efnb3(-/-) or Reln(+/-); Efnb2(-/-)) and triple ephrin B1, B2, B3 knockouts show neuronal migration defects that recapitulate the ones observed in the neocortex, hippocampus and cerebellum of the reeler mouse. Mechanistically, we show that Reelin binds to the extracellular domain of ephrin Bs, which associate at the membrane with VLDLR and ApoER2 in neurons. Clustering of ephrin Bs leads to the recruitment and phosphorylation of Dab1 which is necessary for Reelin signalling. Conversely, loss of function of ephrin Bs severely impairs Reelin-induced Dab1 phosphorylation. Importantly, activation of ephrin Bs can rescue the reeler neuronal migration defects in the absence of Reelin protein. Together, our results identify ephrin Bs as essential components of the Reelin receptor/signalling pathway to control neuronal migration during the development of the nervous system.

An effect of 11-ketotestosterone (11-KT) on growth of previtellogenic (PV) ovaries of eel, salmon and Atlantic cod has been demonstrated. The purpose of this study was to investigate the effects of 11-KT treatment (in vivo) on ovarian growth, on hormonal and biochemical changes in blood, and on ovarian mRNA levels of lipidation-related genes in captive beluga with PV oocytes. In addition, the potential involvement of lipoprotein lipase (Lpl), an important enzyme for extracellular hydrolysis of lipoprotein-associated lipids, was evaluated. Twelve beluga (4-year olds) were treated with an intraperitoneal slow-release implant of either 11-KT (2.5 mg) or a compressed matrix (control). Ovarian biopsy was done to obtain pre- (day 0: TO) and post-treatment (day 21: T21) data on histology and target gene expression. Three weeks of exposure resulted in an increase in serum 11-KT levels from 2.2 ng/mL to 83 ng/mL but did not yield significant changes in serum levels of triacylglycerides and cholesterol. Furthermore, 11-KT implantation increased oocyte diameters from 259 pm (TO) to 309 pm by T21. Regardless of the increase in oocyte size, ovaries remained in the PV stage, mostly as late perinucleolar oocytes. Meanwhile, at the molecular level, the expression of lipidation-related transcripts [lpl, apolipoprotein E (apoe), very low density lipoprotein receptors (vldlr), low-density lipoprotein receptor-related protein 8-like (lrp8)] was significantly up-regulated after three weeks. Immunostaining for Lpl by Western blotting indicated three immunoreactive bands (70, 58 and 37 kDa) in ovarian homogenates from beluga, but signal intensity was not affected by treatment. Altogether, the administration of 11-KT increased 11-KT serum levels, oocyte size, and the expression of genes associated with lipid uptake. However, this treatment did not advance ovarian development beyond the PV stage.