Creating a “Hopeful Monster”: Mouse Forward Genetic Screens

Horner, Vanessa L.; Caspary, Tamara

doi:10.1007/978-1-61779-210-6_12

Cited by 12 publications

(10 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ENU screen, mouse strains and genotyping ENU mutagenesis was performed as previously described (Horner and Caspary, 2011) on Mus musculus C57BL/6J male mice (Jackson Laboratory stock #000664). FVB/NJ females (Jackson Laboratory stock #001800) were used to outcross mutagenized males and their progeny for more than ten generations.…”

Section: Methodsmentioning

confidence: 99%

The Secretory Pathway Calcium ATPase 1 (SPCA1) controls neural tube closure by regulating cytoskeletal dynamics

Brown

García-García

2018

Development

View full text Add to dashboard Cite

Neural tube closure relies on the apical constriction of neuroepithelial cells. Research in frog and fly embryos has found links between the levels of intracellular calcium, actomyosin dynamics and apical constriction. However, genetic evidence for a role of calcium in apical constriction during mammalian neurulation is still lacking. Secretory pathway calcium ATPase (SPCA1) regulates calcium homeostasis by pumping cytosolic calcium into the Golgi apparatus. Loss of function in causes cranial exencephaly and spinal cord defects in mice, phenotypes previously ascribed to apoptosis. However, our characterization of a novel allele of revealed that neurulation defects in mutants are not due to cell death, but rather to a failure of neuroepithelial cells to apically constrict. We show that SPCA1 influences cell contractility by regulating myosin II localization. Furthermore, we found that loss of disrupts actin dynamics and the localization of the actin remodeling protein cofilin 1. Taken together, our results provide evidence that SPCA1 promotes neurulation by regulating the cytoskeletal dynamics that promote apical constriction and identify cofilin 1 as a downstream effector of SPCA1 function.

show abstract

Section: Methodsmentioning

confidence: 99%

The Secretory Pathway Calcium ATPase 1 (SPCA1) controls neural tube closure by regulating cytoskeletal dynamics

Brown

García-García

2018

Development

View full text Add to dashboard Cite

show abstract

“…Because mutagenesis was carried out on B6 males, ENU-induced mutations should be linked to B6 variants for DNA sequences that differ between the B6 and FVB strains. Moreover, all ketu-homozygous G3 males should be homozygous for at least some of the same linked B6 variants (Caspary, 2010;Horner and Caspary, 2011). We therefore roughly mapped the ketu mutation by hybridizing genomic DNA from five G3 mutants to mouse SNP genotyping arrays and searching for genomic regions where all five mice shared homozygosity for B6 SNPs (Caspary, 2010;Jain et al, 2017).…”

Section: Ketu Maps To a Missense Mutation In The Ythdc2 Genementioning

confidence: 99%

ketumutant mice uncover an essential meiotic function for the ancient RNA helicase YTHDC2

Jain

Puno

Meydan³

et al. 2017

Preprint

View full text Add to dashboard Cite

Mechanisms regulating mammalian meiotic progression are poorly understood. Here we identify mouse YTHDC2 as a critical component. A screen yielded a sterile mutant, "ketu", caused by a Ythdc2 missense mutation. Mutant germ cells enter meiosis but proceed prematurely to aberrant metaphase and apoptosis, and display defects in transitioning from spermatogonial to meiotic gene expression programs. ketu phenocopies mutants lacking MEIOC, a YTHDC2 partner. Consistent with roles in post-transcriptional regulation, YTHDC2 is cytoplasmic, has 3ʹ→5ʹ RNA helicase activity in vitro, and has similarity within its YTH domain to an N 6 -methyladenosine recognition pocket. Orthologs are present throughout metazoans, but are diverged in nematodes and, more dramatically, Drosophilidae, where Bgcn is descended from a Ythdc2 gene duplication. We also uncover similarity between MEIOC and Bam, a Bgcn partner unique to schizophoran flies. We propose that regulation of gene expression by YTHDC2-MEIOC is an evolutionarily ancient strategy for controlling the germline transition into meiosis. Version 2: The following experimental changes were incorporated:• RNA-seq analyses of wild-type and Ythdc2 mutant testes at 8, 9, and 10 dpp (i.e., during the germline transition into meiosis).• Purification and characterization of recombinant YTHDC2 protein, demonstrating RNA helicase activity and effect of the ketu mutation.• Expanded histological and cytological analyses (DMC1-staining; PAS-staining and TUNEL assay at 8 dpp; further pH3 and tubulin staining along with SYCP3 staining in abnormal metaphases to evaluate premature metaphase entry; BrdU incorporation to evaluate premeiotic DNA replication; and more complete evaluation of persistent CCNA2 expression), including quantification of stainings at multiple ages (pH3, α-tubulin, BrdU, SYCP3, and CCNA2 stainings as well as TUNEL assay). Cem Meydan, Nathalie Lailler, Christopher E. Mason, and Christopher D. Lima were added as coauthors.

show abstract

“…For some of these such as the scurfy mouse with a regulatory T cell defect due to a mutation in the FoxP3 gene [13], corresponding mutations and disease phenotypes exist in humans. The spontaneous mutation rate can be enhanced by administration of the mutagen N-ethyl-N-nitrosourea (ENU) [14]. ENU induces germline point mutations at a rate of about 1 in 10 6 base pairs [15].…”

Section: Forward Genetic Approaches In Micementioning

confidence: 99%

After GWAS: mice to the rescue?

Ermann

Glimcher

2012

Current Opinion in Immunology

View full text Add to dashboard Cite

The genetics of human autoimmune diseases remain incompletely understood, despite significant progress from genome-wide association studies (GWAS). In this review we outline how mouse studies may help filling these knowledge gaps. Forward genetic approaches including mutagenesis screens and quantitative trait locus mapping can be used to identify candidate genes for in depth analysis in human patient populations. Reverse genetic approaches utilize genetically engineered mice to analyze the function of disease-associated genes and their variants. Inbred strains are a distinctive feature of mouse genetics and we discuss their history, advantages and disadvantages. Three factors need to be considered when comparing experimental results from studies in mice and humans. In addition to species-specific differences, phenotypes are affected by the genetic background of the mouse strain being analyzed, as well as by microbial factors. Despite of these complexities, mice are essential discovery tools in the post GWAS era.

show abstract

Creating a “Hopeful Monster”: Mouse Forward Genetic Screens

Cited by 12 publications

References 61 publications

The Secretory Pathway Calcium ATPase 1 (SPCA1) controls neural tube closure by regulating cytoskeletal dynamics

The Secretory Pathway Calcium ATPase 1 (SPCA1) controls neural tube closure by regulating cytoskeletal dynamics

ketumutant mice uncover an essential meiotic function for the ancient RNA helicase YTHDC2

After GWAS: mice to the rescue?

Contact Info

Product

Resources

About