Search bioRxiv⌕ Search

Biology subjects

Rusch, J.

Publications and source records attributed to Rusch, J..

4 recordsLinked to original sources

Fat cadherin cleavage releases a transcriptionally active nuclear fragment to regulate target gene expression

The conserved atypical cadherin fat (ft) controls cellular processes such as growth, planar cell polarity, and mitochondrial function, in organisms ranging from fruit flies to mammals. Working at the apical-junctional plasma membrane the intracellular domain of the Ft protein, FtICD, binds to and regulates components of the Hippo and PCP pathways. Unexpectedly, we show that FtICD is present in the nucleus in cultured cells as well as in embryonic and larval tissues, and identify nuclear localization and nuclear export signals in FtICD required for this localization. We show that membrane-bound FtICD is cleaved and enters nuclei in vivo. Using endogenously tagged Ft as well as overexpressed FtICD we conducted ChIP-seq experiments and identified putative Ft targets including genes involved in signaling pathways, chromatin organization, pattern formation, and neural development. RNAseq demonstrates that some of these genes are differentially regulated in ft mutants. We observe strong correlations of Ft binding regions with peaks for other factors such as DREF and BEAF-32, as well as the Hpo pathway components Yorkie (Yki) and Scalloped (Sd), suggesting that Ft may act in conjunction with these factors to regulate gene expression. Supporting this hypothesis, we found that Ft can physically interact with both Yki and Sd in co-immunoprecipitation experiments in S2 cells. We propose that the modulation of Hippo pathway activity constitutes one of the nuclear functions of Ft, complementing its established function as an upstream regulator of Hippo signaling.

developmental biology↗

A Comparison of Techniques to Determine Active Motor Threshold for Quadriceps Transcranial Magnetic Stimulation Research

The determination of active motor threshold (AMT) is a critical step in transcranial magnetic stimulation (TMS) research protocols involving voluntary muscle contractions. As AMT is frequently determined using an absolute electromyographic (EMG) threshold (e.g., 200{micro}V peak-to-peak amplitude), wide variation in EMG recordings across participants has given reason to consider a relative threshold (e.g., = 2x background EMG) for AMT determination. However, these approaches have not been systemically compared. PURPOSE: We sought to compare the AMT estimations derived from absolute and relative criteria commonly used to determine AMT in the quadriceps muscles, and assess the test-retest reliability of each approach (absolute = 200{micro}V vs. relative = 2x background EMG). METHODS: Eighteen young adults (9 males and 9 females; mean {+/-} SD age = 23 {+/-} 2 years) visited the research laboratory on two occasions. All testing was conducted on the dominant limb. During each laboratory visit, maximal voluntary isometric contraction (MVIC) quadriceps torque was measured, with all subsequent TMS procedures conducted as participants maintained 10% of MVIC torque. AMT was derived from each criterion using motor evoked potentials recorded from the vastus lateralis (VL) and defined as the lowest stimulator output (SO%) needed to meet the specified criteria within [≥] 5/10 pulses. The order of criteria (i.e., absolute vs. relative) was randomized during the first laboratory visit, and counterbalanced during the second visit. A paired samples t-test, 95% confidence intervals and the effect size were used to compare mean differences in AMT estimations obtained from each criterion during the second laboratory visit. Paired samples t-tests, intraclass correlation coefficients (ICC2,1), standard errors of measurement (SEMs), and the minimal difference (MD) scores were calculated to assess test-retest reliability of each AMT criterion. RESULTS: Differences between the AMT criteria were small and not statistically significant (absolute criterion mean = 48.9%, relative criterion mean = 47.4%; p = .309, Cohens d = 0.247). The absolute criterion demonstrated moderate to excellent reliability (ICC2,1= .866 [0.648 - 0.950], SEM = 7.9%, MD = 10.4%), but higher AMTs were observed in the second visit compared to the first (p = 0.043). The relative criteria demonstrated good-to-excellent test-retest reliability (ICC2,1= .894 [0.746 - 0.959], SEM = 6.9%, MD = 8.9%) and AMTs were not different between visits (p = 0.420). CONCLUSIONQuantifying AMT with an absolute voltage threshold of 200{micro}V peak-to-peak amplitude and a relative voltage threshold 2x background EMG resulted in similar estimations within a single testing session. However, the relative voltage criterion demonstrated superior test-retest reliability. TMS researchers aiming to track corticospinal characteristics across visits should consider implementing relative criterion approaches during their AMT determination protocol.

neuroscience↗

Unified single-cell analysis of testis gene regulation and pathology in 5 mouse strains

By removing the confounding factor of cellular heterogeneity, single cell genomics can revolutionize the study of development and disease, but methods are needed to simplify comparison among individuals. To develop such a framework, we assayed the transcriptome in 62,600 single cells from the testes of wildtype mice, and mice with gonadal defects due to disruption of the genes Mlh3, Hormad1, Cul4a or Cnp. The resulting expression atlas of distinct cell clusters revealed novel markers and new insights into testis gene regulation. By jointly analysing mutant and wildtype cells using a model-based factor analysis method, SDA, we decomposed our data into 46 components that identify novel meiotic gene regulatory programmes, mutant-specific pathological processes, and technical effects. Moreover, we identify, de novo, DNA sequence motifs associated with each component, and show that SDA can be used to impute expression values from single cell data. Analysis of SDA components also led us to identify a rare population of macrophages within the seminiferous tubules of Mlh3-/- and Hormad1-/- testes, an area typically associated with immune privilege. We provide a web application to enable interactive exploration of testis gene expression and components at http://www.stats.ox.ac.uk/~wells/testisAtlas.html

genomics↗

The complement system supports normal postnatal development and gonadal function in both sexes

Male and female infertility are clinically managed and classified as distinct diseases, and relatively little is known about mechanisms of gonadal function common to both sexes. We used genome-wide genetic analysis on 74,896 women and men to find rare genetic variants that modulate gonadal function in both sexes. This uncovered an association with variants disrupting CSMD1, a complement regulatory protein located on 8p23, in a genomic region with an exceptional evolution. We found that Csmd1 knockout mice display a diverse array of gonadal defects in both sexes, and in females, impaired mammary gland development that leads to increased offspring mortality. The complement pathway is significantly disrupted in Csmd1 mice, and further disruption of the complement pathway from joint inactivation of C3 leads to more extreme reproductive defects. Our results can explain a novel human genetic association with infertility and implicate the complement system in the normal development of postnatal tissues.

developmental biology↗