Project description:We report here that gentic methylation in the male germline, from meiocytes to sperm, is established by siRNAs transcribed from transposons with imperfect sequence homology. These siRNAs are synthesized by meiocyte nurse cells (tapetum) via activity of the chromatin remodeler CLASSY3, which is specifically expressed in tapetal cells. Plants that produce siRNAs only in the tapetum have broadly normal DNA methylation of the entire germline. Finally, we also report that these nurse cell-derived siRNAs (niRNAs) silence germline transposons, thereby safeguarding genome integrity. Our results reveal the crucial role of tapetal niRNAs in germline methylation reprogramming, which is remarkably analogous to piRNA-mediated reprogramming in animal germlines.
Project description:The aim of this work was to extend the previous gene expression analysis ofM- M- nurse and forager gene expression, but using the Apis oligo-array designed from the recently sequenced genome.M- M- This study included 28000 features which included ~11000 predicted genes.M- M- The intial study was done using a microarray design based on PCR products of 5500 features derived from an expressed sequence tag analysis of a brain cDNA library.M- M- The variables studied were age (young and old), cast (nurse and forager) and differential gene expression.
Project description:The death and clearance of nurse cells is a consequential milestone in Drosophila melanogaster oogenesis. In preparation for oviposition, the germline-derived nurse cells bequeath to the developing oocyte all their cytoplasmic contents and undergo programmed cell death. The death of the nurse cells is controlled non-autonomously and is precipitated by epithelial follicle cells of somatic origin acquiring a squamous morphology and acidifying the nurse cells externally. Alternatively, stressors such as starvation can induce the death of nurse cells earlier in mid-oogenesis, manifesting apoptosis signatures, followed by their engulfment by epithelial follicle cells. To identify and contrast the molecular pathways underlying these morphologically and genetically distinct cell death paradigms, both mediated by follicle cells, we compared their genome-wide translational profiles before and after differentiating to acquire a phagocytic capability, as well as during well-fed and nutrient-deprived conditions. By coupling the GAL4-UAS system to Translating Ribosome Affinity Purification (TRAP-seq) we performed high-throughput screens to identify pathways selectively activated or repressed by follicle cells to employ nurse cell-clearance routines contextually and preferentially. Our analyses and in vivo follow-up studies identified several key genes and pathways that play vital roles during the making of a healthy egg, such as maintaining the oocyte reserve, ensuring structural integrity of the egg chamber, and regulating metabolic and signaling pathways.
Project description:Monocytes and matching culture-derived nurse-like cells were compared using Illumina's EPIC/850k arrays Monocytes isolated using MACS and NLCs isolated using cytometry sorting as CD14+/CD19-
Project description:The study of the roles of macrophages in the microenvironment of cancer cells (tumor-associated macrophages, TAM) has gained deep insight over the recent years. Here, we describe gene expression profile of chronic lymphocytic leukemia (CLL)-associated macrophages, also called nurse-like cells (NLC), derived from in vitro co-cultures system. We define these NLC as M2-oriented, TAM-like specific subset of macrophages, with very few inter-individual variations in gene expression profiles.
Project description:LC-MS/MS proteomics was used to identify immune proteins in the plasma of the nurse shark (Ginglymostoma cirratum), using a de novo multi-tissue transcriptome generated for this species. LC-MS/MS was then used to assess the host response to immunization with human serum albumin (HSA) and Complete Freund’s Adjuvant (CFA).