Single cell RNA-sequencing on blood cells from Drosophila lymph glands [scRNA-seq]
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ABSTRACT:
PROVIDER: PRJNA714092 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR13953478_1.fastq.gz | Fastqsanger.gz | |||
| SRR13953478_2.fastq.gz | Fastqsanger.gz | |||
| SRR13953479_1.fastq.gz | Fastqsanger.gz | |||
| SRR13953479_2.fastq.gz | Fastqsanger.gz | |||
| SRR13953480_1.fastq.gz | Fastqsanger.gz |
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Project description:The lymph gland is one of the main larval hematopoietic organin in Drosophila melanogaster. In wnandering third instar larav, it is composed of two pairs of anterior lobes, followed by 2 to 3 pair of posterior lobes. To gain further insight into the gene expression repertoire of Drosophila blood cells, we established the gene expression profiles of third instar larva lymph glands anterior and posterior lobes.
2021-02-20 | GSE152416 | GEO
Project description:Transcriptomic analysis of Drosophila larval lymph glands
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Project description:Transcriptome analysis of Drosophila whole larval lymph glands
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Project description:We report the use of single cell mRNA sequencing to identify new blood cell types, subtypes, genetic markers, and important developmental pathways in the Drosophila lymph gland. We analyzed transcript data for 21,157 cells from wild-type early third instar larvae (90-93 hours after eggs lay at 25 degrees C) which contain reporter constructs that mark progenitors and differentiated cells. We find that lymph gland blood cells can be catorgorized into 9 major clusters. One cluster corrresponds to the niche cells of the posterior signaling center (PSC) that expressed the PSC markers Antennopedia and Collier. Progenitors of the medullary zone (MZ) are found in two distinct clusters (MZ1 and MZ2), both of which express the MZ markers domeless and Tep4. One cluster contains intermediate progenitors of the intermediate zone (IZ), and these express markers such as CG30090, lectin-24A, and MFS3 that we found to be IZ-specific in previous bulk RNA-seq experiments with sorted lymph gland cells. Another cluster (PL) contains plasmatocytes that express NimC1, as well as Hml, Pxn, vkg, and Col4a1. Crystal cells are found in one major population that subclusters into two distinct subclusters (CC1 and CC2) that express the markers lz, Hnt, and proPO.. Another transitory population that we dub proplasmatocytes expresses Hml, Pxn, vkg, and Col4a1, but not PL marker Nimc1 or IZ-markers. One cluster is not well understood, butis highly enriched for cell cycle- and replication stress genes. Pseudotime and Trajectory analysis results in fruther sub-division of these zones based on pseudotime states and aids in building a model of lymph gland development that will be the basis for future studies.
2021-11-03 | GSE168819 | GEO
Project description:We report the use of bulk mRNA sequencing on FACS-sorted blood cells to identify new and unique genetic markers for each major differentiated cell type of the Drosophila lymph gland. We used flow cytomotery in a wild-type genetic background whihc contains two fluorescent reporters that allow for the separation of 3 distinct populations. Plasmatcytes express the markers Hml, Pxn, vkg, Col4a1, and NimC1 , while crystal cells are markered by expression of hnt, lz, ProPO. mCC has higher expression of ProPO relitive to iCC, but lower expression of Hml and DsRed. We identified novel markers that are differentially expressed in each of the three major differentiated blood cell populations (PL, iCC, and mCC) that have been useful to definetely identify populations in scRNA-Seq and give information about transcript isoforms in each of the distinct populations.
2021-11-03 | GSE168820 | GEO
Project description:Bulk RNA sequencing on sorted blood cells from Drosophila lymph glands [MZ IZ bulk RNA-seq]
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Project description:Bulk RNA sequencing on sorted blood cells from Drosophila lymph glands [PL iCC mCC bulk RNA-Seq]
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Project description:We report the use of bulk mRNA sequencing on FACS-sorted blood cells to identify new and unique genetic markers for each major zone of the Drosophila lymph gland. We used flow cytomotery in a wild-type genetic background whihc contains two fluorescent reporters that allow for the separation of 4 distinct populations. Progenitors of the medullary zone (MZ) are marked by dome-meso-EGFP expression. Differentitating cels of the cortical zone (CZ) are marked by Hml-DsRed. These two sorted populations (MZ and CZ) are positive for the expected zone-specific markers (i.e. Tep4, dome, and shg for MZ and Hml, Pxn, vkg, Col4a1, and NimC1 for plasmatocytes of the CZ), however, the CZ population also expresses crystal cell markers (hnt, lz, ProPO), indicating that it is a mix of plasmatocytes and crystal cells. Intermediate progenitors of the intermediate zone (IZ), which are not well understood and do not have specific markers, but are characterized by simultaneous expression of EGFP and DsRed (double positives) were also sorted. In addtiion, there is a population of lymph gland cells that is double negative for both EGFP and DsRed fluorescence. This double negative population contains a mix of cell types including posterior signaling center (PSC) cells, MZ and CZ cells that have lost fluorescence, late crystal cells that do not express Hml (early CCs do express), and non-blood cells (i.e. dorsal vessel and pericardial cells). These double negatives were excluded from the analysis for these reasons. We analyzed transcript data of these 4 sorted populations in the early third instart larvae (90-96 hours after eggs lay at 25 degrees C). We identified novel markers that are differentially expressed in each of the three major blood cell populations (MZ, IZ, CZ) that have been useful to definetely identify populations in scRNA-Seq and give information about transcript isoforms in each of the distinct populations.
2021-11-03 | GSE168821 | GEO
Project description:The balance between hematopoietic progenitors and differentiated hemocytes is finely tuned during development. In the larval hematopoietic organ of Drosophila, called the lymph gland, the receptor tyrosine kinase Pvr signals from differentiated cells to maintain a pool of undifferentiated progenitors. However, little is known about the processes that support Pvr function. The small GTPase Ral is involved in the regulation of several membrane trafficking events. Drosophila has a single Ral protein, Rala, which has been implicated in the development of various tissues. Here, we investigated the involvement of Rala in the larval fly hematopoietic system. We discovered that the loss of Rala activity phenocopies Pvr loss of function by promoting hemocyte progenitor differentiation. Moreover, using epistasis analysis, we found that the guanine exchange factor RalGPS lies upstream of Rala in this event, whereas the exocyst and Rab11 are acting downstream. Strikingly, the loss of Rala activity leads to a considerable accumulation of Pvr at the plasma membrane, hence suggesting a trafficking defect and reduced Pvr function. Consistent with this hypothesis, Rala loss of function phenotype in the lymph gland is fully suppressed by constitutive STAT activity, which normally mediates Pvr function in the lymph gland. Together, our findings unravel a novel RalGPS-Rala-exocyst-Rab11 axis for the maintenance of lymph gland homeostasis through Pvr.
2020-05-25 | GSE148035 | GEO
Project description:The Drosophila lymph gland (LG) is the larval hematopoietic organ comprised of multipotent prohemocytes and mature hemocytes and has been a valuable model for understanding mechanisms underlying the hematopoiesis and immune responses. There are three types of mature hemocytes in the lymph gland; plasmatocytes, lamellocytes, and crystal cells, all of which are analogous to myeloid-lineage blood cells. To date, the Drosophila hematopoietic system has been primarily defined by classical genetic methods that may limit the promise of its advantage as a model. In this study, we used single-cell RNA sequencing method to comprehensively profile the heterogeneity of developing myeloid hemocytes in the wild-type lymph gland and their transitions upon active immunity caused by wasp infestation. Moreover, we compared hemocytes originated from the embryonic and the lymph gland hematopoiesis and unraveled genetic and cellular diversity of two different ancestries. Finally, hemocytes of the Drosophila lymph gland and human immune cells are contrasted at a transcriptome level, highlighting evolutionary conservations of myeloid cells across species. Overall, our single-cell RNA seq analyses disclose the development of myeloid hemocytes at a single-cell level and provide comparative insights into two different lineages of hemocytes in Drosophila and perspectives on the evolution of myeloid cells, serving as an ample resource for revealing essentials of the hematopoiesis.
2020-07-24 | GSE141273 | GEO