Imaging cell lineage with a synthetic digital recording system

Cell lineage plays a pivotal role in cell fate determination. Chow et al. demonstrate the use of an integrase-based synthetic barcode system called intMEMOIR, which uses the serine integrase Bxb1 to perform irreversible nucleotide edits. Inducible editing either deletes or inverts its target region, thus encoding information in three-state memory elements, or trits, and avoiding undesired recombination events. Using intMEMOIR combined with single-molecule fluorescence in situ hybridization, the authors were able to identify clonal structures as well as gene expression patterns in the fly brain, enabling both clonal analysis and expression profiling with intact spatial information. The ability to visualize cell lineage relationships directly within their native tissue context provides insights into development and disease

Imaging cell lineage with a synthetic digital recording system

Multicellular development depends on the differentiation of cells into specific fates with precise spatial organization. Lineage history plays a pivotal role in cell fate decisions, but is inaccessible in most contexts. Engineering cells to actively record lineage information in a format readable in situ would provide a spatially resolved view of lineage in diverse developmental processes. Here, we introduce a serine integrase-based recording system that allows in situ readout, and demonstrate its ability to reconstruct lineage relationships in cultured stem cells and flies. The system, termed intMEMOIR, employs an array of independent three-state genetic memory elements that can recombine stochastically and irreversibly, allowing up to 59,049 distinct digital states. intMEMOIR accurately reconstructed lineage trees in stem cells and enabled simultaneous analysis of single cell clonal history, spatial position, and gene expression in Drosophila brain sections. These results establish a foundation for microscopy-readable clonal analysis and recording in diverse systems

Effects of a Single Exercise Bout on Insulin Sensitivity in Black and White Individuals

Background: Previous research suggests non-Hispanic blacks (blacks) are more insulin resistant than non-Hispanic whites (whites). Physical activity can play an important role in reducing insulin resistance. However, it is unknown whether racial differences exist in response to exercise. Therefore, the purpose of this study was to compare metabolic responses to a single bout of exercise in blacks and age-, sex-, and body mass index-matched whites. Methods: Whole-body insulin sensitivity, glucose storage, glucose oxidation, and respiratory exchange ratio (RER) were assessed during a hyperinsulinemic-euglycemic clamp in normoglycemic blacks (n = 11) and whites (n = 10). Outcome measures were evaluated in a sedentary control condition and 12 h after treadmill walking at 75% of maximal heart rate for 75 min. Results: In the control condition, there were no differences in insulin sensitivity between blacks and whites (P = 0.54). During the clamp, glucose oxidation and insulin-stimulated RER values were significantly higher in blacks compared with whites (P = 0.04 and P \u3c 0.01, respectively). Despite similar RER values during exercise, RER values at 60, 90, and 120 min after exercise in blacks were also significantly higher compared with whites (P \u3c 0.05). After exercise, there were no significant improvements in insulin sensitivity (P = 0.57) or glucose storage (P = 0.42) in blacks or whites; however, glucose oxidation was significantly lower in both racial groups (P \u3c 0.05). Conclusions: These data suggest that insulin sensitivity is similar in blacks and age-, sex-, and body mass index-matched whites, but the glucose disposal pathways (storage vs. oxidation) are somewhat different. Compared with whites, blacks appear to have a greater capacity to increase glucose oxidation immediately after exercise and during insulin stimulation

Unraveling structural and magnetic information during growth of nanocrystalline SrFe12O19

[EN] The hydrothermal synthesis of magnetic strontium hexaferrite (SrFe12O19) nanocrystallites was followed in situ using synchrotron powder X-ray diffraction. For all the studied temperatures, the formation of SrFe12O19 happened through an intermediate crystalline phase, identified as the so-called six-line ferrihydrite (FeOOH). The presence of FeOOH has been overlooked in previous studies on hydrothermally synthesized SrFe12O19, despite the phase having a non-trivial influence on the magnetic properties of the final material. The chemical synthesis was successfully reproduced ex situ in a custom-designed batch-type reactor that resembles the experimental conditions of the in situ setup, while allowing larger quantities of material to be produced. The agreement in phase composition between the two studies reveals comparability between both experimental setups. Hexagonal platelet morphology is confirmed for SrFe12O19 combining Rietveld refinements of powder X-ray diffraction (PXRD) data with transmission electron microscopy (TEM). Room temperature magnetization curves were measured on the nanopowders prepared ex situ. The magnetic properties are discussed in the context of the influence of phase composition and crystallite size.The authors would like to thank financial support from the European Commission through the project NANOPYME FP7-NMP-2012-SMALL-6 NANOPYME (No. 310516). The work was likewise supported by the Danish National Research Foundation (Center for Materials Crystallography, DNRF93) and the Danish Research Council for Nature and Universe (Danscatt). Furthermore, support by the European Community's Seventh Framework Programme (FP7/2007-2013) CALIPSO under Grant Agreement No. 312284 is also acknowledged. Support is also gratefully accepted from the Danish Research Council for Technology and Production Sciences through a Sapere Aude grant (Improved Permanent Magnets through Nanostructuring).Peer reviewe

Psychological Responses to Acute Exercise in Sedentary Black and White Individuals

Background: Racial differences in psychological determinants of exercise exist between non-Hispanic blacks (blacks) and non-Hi.spanic whites (whites). To date, no study has examined racial differences in the psychological responses during and after exercise. The objective of tbis study was to compare psychological outcomes of single exercise bouts in blacks and whites. Methods: On 3 separate occasions, sedentary black (n = 16) and white (n = 14) participants walked on a treadmill at 75%max tm for 75 minutes. Questionnaires assessing mocxi, state anxiety, and exercise task self-efficacy were administered before and after each exercise bout. In-task mood and rating of perceived exertion (RPE) were measured every 5 minutes during exercise. Results: Exercise self-efficacy and psychological distress significantly improved in both blacks and whites. However during exercise blacks reported more positive in-task mood and lower RPE compared with whites. Conclusions: These data suggest that racial differences exist in psychological responses during exercise. Further research should confirm these findings in a larger, free-living populatio

A neuropeptide speeds circadian entrainment by reducing intercellular synchrony.

Shift work or transmeridian travel can desynchronize the body's circadian rhythms from local light-dark cycles. The mammalian suprachiasmatic nucleus (SCN) generates and entrains daily rhythms in physiology and behavior. Paradoxically, we found that vasoactive intestinal polypeptide (VIP), a neuropeptide implicated in synchrony among SCN cells, can also desynchronize them. The degree and duration of desynchronization among SCN neurons depended on both the phase and the dose of VIP. A model of the SCN consisting of coupled stochastic cells predicted both the phase- and the dose-dependent response to VIP and that the transient phase desynchronization, or "phase tumbling", could arise from intrinsic, stochastic noise in small populations of key molecules (notably, Period mRNA near its daily minimum). The model also predicted that phase tumbling following brief VIP treatment would accelerate entrainment to shifted environmental cycles. We tested this using a prepulse of VIP during the day before a shift in either a light cycle in vivo or a temperature cycle in vitro. Although VIP during the day does not shift circadian rhythms, the VIP pretreatment approximately halved the time required for mice to reentrain to an 8-h shifted light schedule and for SCN cultures to reentrain to a 10-h shifted temperature cycle. We conclude that VIP below 100 nM synchronizes SCN cells and above 100 nM reduces synchrony in the SCN. We show that exploiting these mechanisms that transiently reduce cellular synchrony before a large shift in the schedule of daily environmental cues has the potential to reduce jet lag

A neuropeptide speeds circadian entrainment by reducing intercellular synchrony.

Shift work or transmeridian travel can desynchronize the body's circadian rhythms from local light-dark cycles. The mammalian suprachiasmatic nucleus (SCN) generates and entrains daily rhythms in physiology and behavior. Paradoxically, we found that vasoactive intestinal polypeptide (VIP), a neuropeptide implicated in synchrony among SCN cells, can also desynchronize them. The degree and duration of desynchronization among SCN neurons depended on both the phase and the dose of VIP. A model of the SCN consisting of coupled stochastic cells predicted both the phase- and the dose-dependent response to VIP and that the transient phase desynchronization, or "phase tumbling", could arise from intrinsic, stochastic noise in small populations of key molecules (notably, Period mRNA near its daily minimum). The model also predicted that phase tumbling following brief VIP treatment would accelerate entrainment to shifted environmental cycles. We tested this using a prepulse of VIP during the day before a shift in either a light cycle in vivo or a temperature cycle in vitro. Although VIP during the day does not shift circadian rhythms, the VIP pretreatment approximately halved the time required for mice to reentrain to an 8-h shifted light schedule and for SCN cultures to reentrain to a 10-h shifted temperature cycle. We conclude that VIP below 100 nM synchronizes SCN cells and above 100 nM reduces synchrony in the SCN. We show that exploiting these mechanisms that transiently reduce cellular synchrony before a large shift in the schedule of daily environmental cues has the potential to reduce jet lag

Data for 'Imaging cell lineage with a synthetic digital recording system'

This deposit contains the data, code, and analysis to recreate the results in the manuscript, 'Imaging cell lineage with a synthetic digital recording system', and the sequence information of relevant constructs. The datasets are organized by figures, and the codes are written for MatLab, R, and Python. Please see the related publication for more details, and contact the corresponding authors for any questions.Related Publication:</p> Imaging cell lineage with a synthetic digital recording system</p> https://doi.org/10.1101/2020.02.21.958678</p>en