A new mouse line for cell ablation by diphtheria toxin subunit A controlled by a Cre‐dependent FLEx switch

Plummer, Nicholas W.; Ungewitter, Erica; Smith, Kathleen G.; Yao, Hisayuki; Jensen, Patricia

doi:10.1002/dvg.23067

Cited by 15 publications

(8 citation statements)

References 33 publications

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“…To test this hypothesis, we depleted macrophages in vivo by crossing transgenic mice, expressing codon-optimised Cre (iCre) under the control of the Csf1r promoter ( Csf1r icre ), with transgenic mice that had a floxed-STOP cassette and the diphtheria toxin A subunit (DTA) knocked into the Rosa26 locus ( Rosa DTA ) (Figure 1—figure supplement 2). In this system, DTA expression is specifically induced in iCre + cells, resulting in cell death by inhibiting protein synthesis (Breitman et al, 1987; Collier, 2001; Plummer et al, 2017) (Figure 1—figure supplement 2). Consistent with the hypothesis that macrophages clear rostral nephrogenic progenitors, macrophage-depleted E11.5 Csf1ricre icre+ Rosa DTA embryos had elongated metanephric mesenchyme populations and larger rostral clusters of Six2 + cells compared to somite pair-matched littermate controls (Figure 1i–l).…”

Section: Resultsmentioning

confidence: 99%

Macrophages restrict the nephrogenic field and promote endothelial connections during kidney development

Munro

Wineberg

Tarnick

et al. 2019

eLife

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The origins and functions of kidney macrophages in the adult have been explored, but their roles during development remain largely unknown. Here we characterise macrophage arrival, localisation, heterogeneity, and functions during kidney organogenesis. Using genetic approaches to ablate macrophages, we identify a role for macrophages in nephron progenitor cell clearance as mouse kidney development begins. Throughout renal organogenesis, most kidney macrophages are perivascular and express F4/80 and CD206. These macrophages are enriched for mRNAs linked to developmental processes, such as blood vessel morphogenesis. Using antibody-mediated macrophage-depletion, we show macrophages support vascular anastomoses in cultured kidney explants. We also characterise a subpopulation of galectin-3+ (Gal3+) myeloid cells within the developing kidney. Our findings may stimulate research into macrophage-based therapies for renal developmental abnormalities and have implications for the generation of bioengineered kidney tissues.

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Section: Resultsmentioning

confidence: 99%

Macrophages restrict the nephrogenic field and promote endothelial connections during kidney development

Munro

Wineberg

Tarnick

et al. 2019

eLife

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“…In addition, because the opto- or chemogenetic tool is expressed in a conditional manner, it is only expressed by cells of a specific genotype, thus giving the investigator both neuroanatomical and neurochemical control over the response. Because the receptor transcript is packaged within a Flip-Excision-Switch (FLEX) cassette, the functional receptor can be expressed only in the presence of cre-recombinase (Schnutgen et al, 2003; Fuller et al, 2015; Plummer et al, 2017). The use of the Cre-driver mouse lines, where cre is expressed downstream of a selected promoter, ensures that the designer receptors when injected in these mice are expressed in a Cre-dependent manner, specifically in neurons that express a select protein.…”

Section: Genetic Tools and Technologies For Circuit Analysismentioning

confidence: 99%

Neural Circuitry Underlying Waking Up to Hypercapnia

Kaur

Saper

2019

Front. Neurosci.

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Obstructive sleep apnea is a sleep and breathing disorder, in which, patients suffer from cycles of atonia of airway dilator muscles during sleep, resulting in airway collapse, followed by brief arousals that help re-establish the airway patency. These repetitive arousals which can occur hundreds of times during the course of a night are the cause of the sleep-disruption, which in turn causes cognitive impairment as well as cardiovascular and metabolic morbidities. To prevent this potential outcome, it is important to target preventing the arousal from sleep while preserving or augmenting the increase in respiratory drive that reinitiates breathing, but will require understanding of the neural circuits that regulate the cortical and respiratory responses to apnea. The parabrachial nucleus (PB) is located in rostral pons. It receives chemosensory information from medullary nuclei that sense increase in CO2 (hypercapnia), decrease in O2 (hypoxia) and mechanosensory inputs from airway negative pressure during apneas. The PB area also exerts powerful control over cortical arousal and respiration, and therefore, is an excellent candidate for mediating the EEG arousal and restoration of the airway during sleep apneas. Using various genetic tools, we dissected the neuronal sub-types responsible for relaying the stimulus for cortical arousal to forebrain arousal circuits. The present review will focus on the circuitries that regulate waking-up from sleep in response to hypercapnia.

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“…To determine whether the loss of a subset of touch neurons implicated in social touch would impact development or adult behavior, we crossed Mrprb4 Cr e mice to Rosa DTA mice to genetically ablate Mrgprb4 neurons (Fig1A). Because Mrgprb4 expression begins around postnatal day (PND 2) 19 , and the cre-dependent ablation method ablates cells ∼1 day after Cre expression starts 23 , Mrgprb4 lineage neurons were likely ablated ∼PND 3. Using RNAscope in situ hybridization, we confirmed the extinction of cells that express Mrgprb4 (Fig 1B-C).…”

Section: Resultsmentioning

confidence: 99%

A critical role for touch neurons in a skin-brain pathway for stress resilience

Schaffler

Johnson

Hing

et al. 2022

Preprint

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Social touch can act as a stress buffer, reducing behavioral and physiological responses to stressful scenarios. However, skin-brain touch pathways that promote stress resilience remain unknown. Here, we show that mice with an early life genetic ablation of Mrgprb4-lineage touch neurons display stress vulnerability behaviors in adulthood. Chemogenetic activation of these touch neurons reduced corticosterone levels under mild acute stress conditions. In addition, whole-brain c-Fos activity mapping while chemogenetically turning on these neurons uncovered differential neural activity patterns in brain areas relevant to somatosensation, reward, and affect. To gain mechanistic insight into this skin-brain touch pathway for stress susceptibility, we used multi-circuit neurophysiological recordings across seven brain regions at baseline and after stress in mice that had Mrgprb4-lineage touch neurons ablated in early life. Interestingly, the Mrgprb4-lineage neuron- ablated mice have alterations in local field potential phase directionality and power in the theta frequencies in mesolimbic reward regions, which may underlie our observed stress susceptibility phenotype. Together, these studies revealed that sensory neurons in the skin engage networks across the brain to promote stress resilience.

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A new mouse line for cell ablation by diphtheria toxin subunit A controlled by a Cre‐dependent FLEx switch

Cited by 15 publications

References 33 publications

Macrophages restrict the nephrogenic field and promote endothelial connections during kidney development

Macrophages restrict the nephrogenic field and promote endothelial connections during kidney development

Neural Circuitry Underlying Waking Up to Hypercapnia

A critical role for touch neurons in a skin-brain pathway for stress resilience

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