Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Aug;31(7-8):205-214.
doi: 10.1007/s00335-020-09847-z. Epub 2020 Aug 29.

Baseline and innate immune response characterization of a Zfp30 knockout mouse strain

Lucas T Laudermilk  1   2 , Adelaide Tovar  1   2 , Alison K Homstad  1   2 , Joseph M Thomas  1 , Kathryn M McFadden  1 , Miriya K Tune  3 , Dale O Cowley  1   4 , Jason R Mock  3 , Folami Ideraabdullah  1   2   5 , Samir N P Kelada  6   7   8
Affiliations

Baseline and innate immune response characterization of a Zfp30 knockout mouse strain

Lucas T Laudermilk et al. Mamm Genome. 2020 Aug.

Abstract

Airway neutrophilia is correlated with disease severity in a number of chronic and acute pulmonary diseases, and dysregulation of neutrophil chemotaxis can lead to host tissue damage. The gene Zfp30 was previously identified as a candidate regulator of neutrophil recruitment to the lungs and secretion of CXCL1, a potent neutrophil chemokine, in a genome-wide mapping study using the Collaborative Cross. ZFP30 is a putative transcriptional repressor with a KRAB domain capable of inducing heterochromatin formation. Using a CRISPR-mediated knockout mouse model, we investigated the role that Zfp30 plays in recruitment of neutrophils to the lung using models of allergic airway disease and acute lung injury. We found that the Zfp30 null allele did not affect CXCL1 secretion or neutrophil recruitment to the lungs in response to various innate immune stimuli. Intriguingly, despite the lack of neutrophil phenotype, we found there was a significant reduction in the proportion of live Zfp30 homozygous female mutant mice produced from heterozygous matings. This deviation from the expected Mendelian ratios implicates Zfp30 in fertility or embryonic development. Overall, our results indicate that Zfp30 is an essential gene but does not influence neutrophilic inflammation in this particular knockout model.

PubMed Disclaimer

Conflict of interest statement

Conflicts of interest:

Dale Cowley is employed by, has equity ownership in and serves on the board of directors of TransViragen, the company which has been contracted by UNC-Chapel Hill to manage its Animal Models Core Facility.

Figures

Figure 1.
Figure 1.. Allele-specific qPCR confirms genotype-dependent expression of Zfp30 wildtype and mutant alleles in the lung.
Primer pairs that specifically amplify either the wildtype or CRISPR-Cas9 modified allele were used to confirm the status of Zfp30 expression in whole lung tissue from 4–6 mice per genotype. Mice were 9–10 weeks of age at harvest.
Figure 2.
Figure 2.. Zfp30−/− mice exhibit no differences in glucose tolerance but have lower body weight.
A. Thirteen week old mice were fasted overnight and dosed with glucose (2 g per kg lean body mass) via intraperitoneal injection. Blood glucose was then measured at baseline and 15, 30, 45, 60, and 120 minutes after injections. B. Bodyweight at harvest. C. Lean body mass determined by MRI. D. Fat mass determined by MRI normalized to body weight. N=10–11 per genotype. * P < 0.05; ^ P < 0.1.
Figure 3.
Figure 3.. No difference in CXCL1 secretion by genotype in primary mouse tracheal epithelial cells after LPS exposure.
Mouse tracheal epithelial cell cultures were generated from Zfp30+/+ and Zfp30−/− mice and maintained at air-liquid interface for 21 days. Cells were then exposed to 100 μM LPS for 24 hours, and supernatants were assayed for CXCL1 concentrations using Luminex assays. N= 6 per condition per genotype.
Figure 4.
Figure 4.. No difference in neutrophil count or CXCL1 concentration in BALF after house dust mite allergen challenge in Zfp30+/+ and Zfp30−/− mice.
Zfp30+/+ and Zfp30−/− mice were sensitized to and challenged with HDM allergen, and effects on neutrophil counts (A) and CXCL1 concentration (B) in BALF were monitored 48 and 72 hours after challenge. Cell count results are the combined data of two experiments and were assayed via differential cell counts. Cytokine were measured using multiplex assays. N= 9–11 mice per condition per genotype.
Figure 5.
Figure 5.. No differences in neutrophil chemotaxis by Zfp30 genotype in two models of neutrophilic inflammation.
(A-C) Zfp30+/+ and Zfp30−/− mice were given LPS by intratracheal administration and phenotyped 48, 72, or 96 hours later Neutrophils counts (A) and neutrophils as a percentage of total cell count in bronchoalveolar lavage (B) were we measured at indicated time points. CXCL1 was measured at 48 hours post-challenge (C). Neutrophil counts reported are from a single intratracheal administration and are representative of four experiments that span collections 8–96 hours after challenge. Neutrophil levels were assayed via differential cell counts. N= 8 per genotype per condition. Cytokine measurements were carried out via Luminex assays. N= 10 per genotype. (D-E) Zfp30+/+ and Zfp30−/− mice were exposed to 2 ppm ozone for three hours and phenotyped 21 hours later. Total (D) and percent neutrophils (E) are shown. N= 4–6/genotype/group.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Charo IF, and Ransohoff RM, 2006. The Many Roles of Chemokines and Chemokine Receptors in Inflammation. N. Engl. J. Med 354: 610–621. - PubMed
    1. Chen W, Schwalie PC, Pankevich EV, et al. ZFP30 promotes adipogenesis through the KAP1-mediated activation of a retrotransposon-derived Pparg2 enhancer. Nat Commun. 2019;10(1):1809. - PMC - PubMed
    1. Denny P, Hopes E, Gingles N, Broman K, McPheat W et al., 2003. A major locus conferring susceptibility to infection by Streptococcus pneumoniae in mice. Mamm. Genome 14: 448–453. - PubMed
    1. Dickinson M, Flenniken A, Ji X et al., 2016. High-throughput discovery of novel developmental phenotypes. Nature 537, 508–514. - PMC - PubMed
    1. Donoghue LJ, Livraghi-Butrico A, McFadden KM, Thomas JM, Chen G et al., 2017. Identification of trans protein QTL for secreted airway mucins in mice and a causal role for Bpifb1. Genetics 207: 801–812. - PMC - PubMed

Publication types

MeSH terms