ABSTRACT
NEDD9 (neural precursor cell expressed, developmentally down-regulated 9) is a member of the CAS (Crk-associated substrate) family of scaffolding proteins that regulate cell adhesion and migration. A Nedd9 knock-out/lacZ knock-in mouse (Nedd9(-/)(-)) was developed in order to study Nedd9 expression and function in the nervous system. Herein we show that NEDD9 is expressed in the adult brain and is prominently expressed in the hippocampus. Behavioral testing uncovered functional deficits in Nedd9(-)(/)(-) mice. In the Morris water maze test, Nedd9(-)(/)(-) mice showed deficits in both the ability to learn the task as well as in their ability to recall the platform location. There was no change in the gross morphology of the hippocampus, and stereological analysis of BrdU-labeled newly formed hippocampal cells suggested that this defect is not secondary to altered neurogenesis. However, analysis of the hippocampus revealed extensive loss of dendritic spine density in both the dentate gyrus (DG) and CA1 regions. Spine loss occurred equally across all branch orders and regions of the dendrite. Analysis of spine density in Nedd9(-)(/)(-) mice at 1.5, 6 and 10 months revealed an age-dependent spine loss. This work shows that NEDD9 is required for the maintenance of dendritic spines in the hippocampus, and suggests it could play a role in learning and memory.
Subject(s)
Adaptor Proteins, Signal Transducing/deficiency , Cognition Disorders/metabolism , Dendritic Spines/metabolism , Hippocampus/metabolism , Animals , Cognition Disorders/pathology , Dendritic Spines/pathology , Female , Gene Knock-In Techniques , Hippocampus/pathology , Male , Mice , Mice, Inbred C57BL , Mice, KnockoutABSTRACT
This study describes the development of an human granulocyte-macrophage colony-stimulating factor DNA cationic-lipid complexed autologous tumour cell vaccine (hGM-CSF CLDC ATCV) and its implementation, following a chemotherapy treatment protocol, in a randomized, placebo-controlled, double-blinded clinical trial in pet dogs with naturally occurring lymphoma. We hypothesized that the use of this vaccine would result in an antitumour immune response leading to improved first remission duration and overall survival in dogs with B-cell lymphoma when compared with chemotherapy alone. Immune stimulation generated by hGM-CSF CLDC ATCV was assessed by means of surrogate in vivo analysis (delayed-type hypersensitivity [DTH]) as well as an ex vivo cellular assay (lymphocyte proliferation assay). The vaccine approach considered in the current report did not result in clinically improved outcomes. A small measure of immunomodulation was documented by DTH and several modifications to the approach are suggested. This report illustrates the feasibility of clinical trials with vaccine strategies using companion animals with non-Hodgkin's lymphoma.
ABSTRACT
Lysozymes are enzymes characterized by their ability to break down bacterial cell walls. In insects certain lysozymes are only found in the midgut, whereas others are only found in the haemolymph and fat body after immune challenge. We identified two lysozyme-encoding cDNAs from Aedes aegypti. Both deduced protein sequences are basic in nature, contain 148 amino acids including eight highly conserved cysteine residues, and their genomic sequences contain a single intron. Transcriptional profiles indicated that the predominant form is constitutively expressed and up-regulated upon immune challenge and blood feeding in adult mosquitoes. The second form is expressed during early developmental stages, larvae and pupae, and at low levels in adults after immune challenge. Lysozymes in Aedes aegypti play both roles, defined by the spatial and temporal regulation of their expression.
