Meiosis is essential for fertility in sexually reproducing species, extensive studies tried to delineate this sophisticated process. Notwithstanding, the molecules involved in meiosis have not been fully characterized. In this study, we investigate the role of zinc finger protein 541 (ZFP541) and its interacting protein potassium channel tetramerization domain containing 19 (KCTD19) in mice. We demonstrate that they are indispensable for male fertility by regulating proper pachytene progression. ZFP541 is expressed starting from leptotene to round spermatids, and KCTD19 is initially expressed in pachytene. Depletion of Zfp541 or Kctd19 leads to infertility in male mice, and exhibits retarded progression from early to mid/late pachynema. In addition, Zfp541 -/ - spermatocytes show abnormal programmed DNA double-strand breaks (DSBs) repair, impaired crossover formation/resolution, and asynapsis of the XY chromosomes. Immunoprecipitation-mass spectrometry (IP-MS) and in vitro Co-IP reveal that ZFP541 interacts with KCTD19, histone deacetylase 1/2 (HDAC1), HDAC2 and deoxynucleotidyltransferase terminal-interacting protein 1 (DNTTIP1). Furthermore, RNA-seq and CUT&Tag analyses demonstrate that ZFP541 binds to the promoter regions of genes involved in meiosis and post-meiosis including Kctd19 , and activates their transcription. Taken together, our studies reveal a ZFP541- Kctd19 transcription regulatory axis and the crucial role of ZFP541 and KCTD19 for pachytene progression and fertility in male mice.

Abstract Background Heterotaxy (Htx) syndrome is an congenital disorders resulting from incorrectly establishment of left-right patterning during embryogenesis. Over 96% of patients with Htx exhibit some form of congenital heart disease (CHD),and has relatively poor survival. Multiple lines of evidence support genetic contributions to the etiology of Htx. As a specific genetic etiology is currently identifiable in only a minority of patients, there remains enormous potential for novel gene and pathway discovery. Purpose The aim of this study was to investigate the diagnostic yield of whole-exome sequencing (WES) and whole-genome sequencing (WGS) in fetuses with the pathogenesis of Htx, to explore candidate genes for Htx and to expand the clinical phenotype of known genetic conditions. Method WES and WGS were performed on specimens from 46 fetuses diagnosed with Htx and their parents. The single-nucleotide variants (SNVs) and copy-number variants (CNVs) were filtered and annotated by standard analysis process. All reported variants were classified according to he American College of Medical Genetics and Genomics guidelines. Results In the 46 fetuses, the detection rates of pathogenic and likely pathogenic variations were21.7% (10/46) and 10.9% (5/46) respectively. Ten pathogenic variations were identified on genes of CCDC114, DNAH11, ARMC4, STRA6, PQBP1 (hemizygote), HYDIN, RAI1 (Alagille Syndrome), ZFMP2 and Del(22q11.2) Syndrome. Five likely pathogenic variation were on DNAAF1 (Holshner syndrome), NF1, NEXN, NOTCH3 and FOXC1. Of 30 fetuses with prenatally diagnosed right atrial isomerism (RAI), the main intracardiac anomalies were atrioventricular canal (AV canal), isomerism of right atrial appendages, pulmonary stenosis or atresia (PS & PA) and right aortic arch. In 16 fetuses diagnosed left atrial isomerism (LAI) the main intracardiac anomalies were isomerism of left atrial appendages, interrupted IVC and azygos vein continuation. Of the 10 positive cases, 8 fetus were diagnosed of RAI and 2 were diagnosed of LAI by prenatal ultrasonic examination or fetal autopsy. The detection rate was 8/30 (26.7%) for RAI and 2/16 (12.5%) for LAI. Conclusion This study outlines the way for a substantial improvement in the diagnostic yield of prenatal genetic disorders in Htx through WES and WGS. Our experience also expanded the knowledge of the clinical phenotype of known genetic conditions. Our results indicate that the proportion of SNV in Htx of prenatal cases was significantly higher than that in patients with other congenital heart abnormalities, and the recessive inheritance occurred in a higher proportion in Htx. Our results have important implications for clinical management and genetic counseling of Htx. Acknowledgement/Funding Ministry of Science and Technology of the People's Republic of China

replying to J. Luo et al. Nature501, 10.1038/nature12589 (2013). We agree with Luo et al . 1 that the magnetoresistance effects that we reported 2 were dependent on the method used to measure them. The reason that there is a difference in the results depending on whether method 1 or method 2 is used (adopting the measurement notation of ref. 1 ) is that there are two voltage-stabilizing diodes in the Keithley 2400 instrument we used. We were unaware that when this instrument was used both as current source and voltmeter, one diode connected the input port of the current source to the input port of the voltmeter, whereas the other diode connected the output port of the current source to the output port of the voltmeter. The diodes caused a crossover of the Hall coefficient from negative to positive when the instrument was used to conduct a Hall measurement in this configuration, leading us to propose an invalid mechanism for the abnormal magnetoresistance. Therefore the mechanism we proposed 2 —minority injection and an induced p-n boundary—does not provide a correct explanation for the observed geometry-enhanced magnetoresistance. Although such a mechanism does not operate in our samples, we note that a p–n boundary could still enhance magnetoresistance in certain circumstances according to our and others’ theoretical calculations and experiments 2 , 3 , 4 .