Genetic tracing of the epithelial lineage during mammalian kidney repair

Developing new therapeutic approaches to treat acute kidney injury requires a detailed understanding of endogenous cellular repair. Genetic fate mapping defines cellular hierarchies in vivo and we used this technique to assess a possible contribution of non-epithelial stem cells to renal repair after ischemic injury. Mice with efficient labeling of renal epithelial cells, but not non-epithelial interstitial cells, were subjected to a single cycle or sequential cycles of kidney injury and repair. No dilution of the epithelial cell fate marker was observed despite robust epithelial cell proliferation. Thus, non-tubular cells do not have the ability to migrate across the basement membrane and differentiate into epithelial cells in this model. Instead, surviving tubular epithelial cells are responsible for repair of the damaged nephron. Future studies will need to distinguish between uniform dedifferentiation and proliferation of all epithelial cells after injury versus selective expansion of an intratubular epithelial stem cell

Genetic tracing of the epithelial lineage during mammalian kidney repair

Developing new therapeutic approaches to treat acute kidney injury requires a detailed understanding of endogenous cellular repair. Genetic fate mapping defines cellular hierarchies in vivo and we used this technique to assess a possible contribution of non-epithelial stem cells to renal repair after ischemic injury. Mice with efficient labeling of renal epithelial cells, but not non-epithelial interstitial cells, were subjected to a single cycle or sequential cycles of kidney injury and repair. No dilution of the epithelial cell fate marker was observed despite robust epithelial cell proliferation. Thus, non-tubular cells do not have the ability to migrate across the basement membrane and differentiate into epithelial cells in this model. Instead, surviving tubular epithelial cells are responsible for repair of the damaged nephron. Future studies will need to distinguish between uniform dedifferentiation and proliferation of all epithelial cells after injury versus selective expansion of an intratubular epithelial stem cell

Resource costs for fault-tolerant linear optical quantum computing

Linear optical quantum computing (LOQC) seems attractively simple: information is borne entirely by light and processed by components such as beam splitters, phase shifters and detectors. However this very simplicity leads to limitations, such as the lack of deterministic entangling operations, which are compensated for by using substantial hardware overheads. Here we quantify the resource costs for full scale LOQC by proposing a specific protocol based on the surface code. With the caveat that our protocol can be further optimised, we report that the required number of physical components is at least five orders of magnitude greater than in comparable matter-based systems. Moreover the resource requirements grow higher if the per-component photon loss rate is worse than one in a thousand, or the per-component noise rate is worse than $10^{-5}$. We identify the performance of switches in the network as the single most influential factor influencing resource scaling

Mouse kidney nuclear isolation and library preparation for single-cell combinatorial indexing RNA sequencing

Single-cell combinatorial indexing RNA sequencing (sci-RNA-seq3) enables high-throughput single-nucleus transcriptomic profiling of multiple samples in one experiment. Here, we describe an optimized protocol of mouse kidney nuclei isolation and sci-RNA-seq3 library preparation. The use of a dounce tissue homogenizer enables nuclei extraction with high yield. Fixed nuclei are processed for sci-RNA-seq3, and self-loaded transposome Tn5 is used for tagmentation in library generation. The step-by-step protocol allows researchers to generate scalable single-cell transcriptomic data with common laboratory supplies at low cost. For complete details on the use and execution of this protocol, please refer to Li et al. (2022)

Lineage-tracing methods and the kidney

The kidney is a complex organ with over 30 different cell types, and understanding the lineage relationships between these cells is challenging. During nephrogenesis, a central question is how the coordinated morphogenesis, growth, and differentiation of distinct cell types leads to development of a functional organ. In mature kidney, understanding cell division and fate during injury, regeneration and aging are critical topics for understanding disease. Genetic lineage tracing offers a powerful tool to decipher cellular hierarchies in both development and disease because it allows the progeny of a single cell, or group of cells, to be tracked unambiguously. Recent advances in this field include the use of inducible recombinases, multicolor reporters, and mosaic analysis. In this review, we discuss lineage-tracing methods focusing on the mouse model system and consider the impact of these methods on our understanding of kidney biology and prospects for future application

Chromatin conformation and histone modification profiling across human kidney anatomic regions

The three major anatomic regions of the human kidney include the cortex, medulla and papilla, with different functions and vulnerabilities to kidney diseases. Epigenetic mechanisms underlying these anatomic structures are incompletely understood. Here, we performed chromatin conformation capture with Hi-C and histone modification H3K4me3/H3K27me3 Cleavage Under Targets and Release Using Nuclease (CUT&RUN) sequencing on the kidney cortex, medulla and papilla dissected from one individual donor. Nuclear suspensions were generated from each region and split subjected to paired Hi-C and CUT&RUN sequencing. We evaluated the quality of next-generation sequencing data, Hi-C chromatin contact matrices and CUT&RUN peak calling. H3K4me3 and H3K27me3 histone modifications represent active and repressive gene transcription, respectively, and differences in chromatin conformation between kidney regions can be analyzed with this dataset. All raw and processed data files are publicly available, allowing researchers to survey the epigenetic landscape across regional human kidney anatomy

An Optimal Design for Universal Multiport Interferometers

Universal multiport interferometers, which can be programmed to implement any linear transformation between multiple channels, are emerging as a powerful tool for both classical and quantum photonics. These interferometers are typically composed of a regular mesh of beam splitters and phase shifters, allowing for straightforward fabrication using integrated photonic architectures and ready scalability. The current, standard design for universal multiport interferometers is based on work by Reck et al (Phys. Rev. Lett. 73, 58, 1994). We demonstrate a new design for universal multiport interferometers based on an alternative arrangement of beam splitters and phase shifters, which outperforms that by Reck et al. Our design occupies half the physical footprint of the Reck design and is significantly more robust to optical losses.Comment: 8 pages, 4 figure