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1.
Cell Metab ; 24(4): 593-607, 2016 10 11.
Article in English | MEDLINE | ID: mdl-27667667

ABSTRACT

Hormone-secreting cells within pancreatic islets of Langerhans play important roles in metabolic homeostasis and disease. However, their transcriptional characterization is still incomplete. Here, we sequenced the transcriptomes of thousands of human islet cells from healthy and type 2 diabetic donors. We could define specific genetic programs for each individual endocrine and exocrine cell type, even for rare δ, γ, ε, and stellate cells, and revealed subpopulations of α, ß, and acinar cells. Intriguingly, δ cells expressed several important receptors, indicating an unrecognized importance of these cells in integrating paracrine and systemic metabolic signals. Genes previously associated with obesity or diabetes were found to correlate with BMI. Finally, comparing healthy and T2D transcriptomes in a cell-type resolved manner uncovered candidates for future functional studies. Altogether, our analyses demonstrate the utility of the generated single-cell gene expression resource.


Subject(s)
Diabetes Mellitus, Type 2/genetics , Diabetes Mellitus, Type 2/pathology , Gene Expression Profiling/methods , Islets of Langerhans/metabolism , Single-Cell Analysis/methods , Glucagon-Like Peptide-1 Receptor/metabolism , Humans , Male , Obesity/genetics , Reproducibility of Results , Tissue Donors , Transcription Factors/metabolism
2.
Nat Biotechnol ; 34(2): 199-203, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26689543

ABSTRACT

Despite the importance of the mammalian neocortex for complex cognitive processes, we still lack a comprehensive description of its cellular components. To improve the classification of neuronal cell types and the functional characterization of single neurons, we present Patch-seq, a method that combines whole-cell electrophysiological patch-clamp recordings, single-cell RNA-sequencing and morphological characterization. Following electrophysiological characterization, cell contents are aspirated through the patch-clamp pipette and prepared for RNA-sequencing. Using this approach, we generate electrophysiological and molecular profiles of 58 neocortical cells and show that gene expression patterns can be used to infer the morphological and physiological properties such as axonal arborization and action potential amplitude of individual neurons. Our results shed light on the molecular underpinnings of neuronal diversity and suggest that Patch-seq can facilitate the classification of cell types in the nervous system.


Subject(s)
Gene Expression Profiling/methods , Neurons/physiology , Patch-Clamp Techniques/methods , Sequence Analysis, RNA/methods , Transcriptome/physiology , Animals , Cell Shape/physiology , Female , Male , Mice , Neocortex/cytology , Neurons/cytology , Neurons/metabolism
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