The effect of Dnaaf5 gene dosage on primary ciliary dyskinesia phenotypes

DNAAF5 is a dynein motor assembly factor associated with the autosomal heterogenic recessive condition of motile cilia, primary ciliary dyskinesia (PCD). The effects of allele heterozygosity on motile cilia function are unknown. We used CRISPR-Cas9 genome editing in mice to recreate a human missense variant identified in patients with mild PCD and a second, frameshift-null deletion in Dnaaf5. Litters with Dnaaf5 heteroallelic variants showed distinct missense and null gene dosage effects. Homozygosity for the null Dnaaf5 alleles was embryonic lethal. Compound heterozygous animals with the missense and null alleles showed severe disease manifesting as hydrocephalus and early lethality. However, animals homozygous for the missense mutation had improved survival, with partially preserved cilia function and motor assembly observed by ultrastructure analysis. Notably, the same variant alleles exhibited divergent cilia function across different multiciliated tissues. Proteomic analysis of isolated airway cilia from mutant mice revealed reduction in some axonemal regulatory and structural proteins not previously reported in DNAAF5 variants. Transcriptional analysis of mouse and human mutant cells showed increased expression of genes coding for axonemal proteins. These findings suggest allele-specific and tissue-specific molecular requirements for cilia motor assembly that may affect disease phenotypes and clinical trajectory in motile ciliopathies

7th Drug hypersensitivity meeting: part two

No abstract availabl

A specific transitory increase in intracellular calcium induced by progesterone promotes acrosomal exocytosis in mouse sperm

During capacitation, sperm acquire the ability to undergo the acrosome reaction (AR), an essential step in fertilization. Progesterone produced by cumulus cells has been associated with various physiological processes in sperm, including stimulation of AR. An increase in intracellular Ca2+ ([Ca2+]i) is necessary for AR to occur. In this study, we investigated the spatio- temporal correlation between the changes in [Ca2+]i and AR in single mouse spermatozoa in response to Progesterone. We found that Progesterone stimulates an [Ca2+]i increase in five different patterns: gradual increase, oscillatory, late transitory, immediate transitory and sustained. We also observed that the [Ca2+]i increase promoted by Progesterone starts at either the flagellum or the head. We validated the use of FM4-64 as an indicator for the occurrence of the AR by simultaneously detecting its fluorescence increase and the loss of EGFP in transgenic EGFPAcr sperm. For the first time, we have simultaneously visualized the rise in [Ca2+]i and the process of exocytosis in response to Progesterone and found that only a specific transitory increase in [Ca2+]i originated in the sperm head promotes the initiation of AR.Fil: Romarowski, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Sánchez Cárdenas, Claudia. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: Ramírez Gómez, Héctor V.. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: Puga Molina, Lis del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Treviño, Claudia L.. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: Hernández Cruz, Arturo. Universidad Nacional Autónoma de México. Instituto de Fisiología Celular; MéxicoFil: Darszon, Alberto I.. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: Buffone, Mariano Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Reconstructions of the Mediterranean Outflow Water during the quaternary based on the study of changes in buried mounded drift stacking pattern in the Gulf of Cadiz

Contourite deposits in the central sector of the middle slope of the Gulf of Cadiz have been studied using a comprehensive acoustic, seismic and core database. Buried, mounded, elongated and separated drifts developed under the influence of the lower core of the Mediterranean Outflow Water are preserved in the sedimentary record. These are characterised by depositional features in an area where strong tectonic and erosive processes are now dominant. The general stacking pattern of the depositional system is mainly influenced by climatic changes through the Quaternary, whereas changes in the depositional style observed in two, buried, mounded drifts, the Guadalquivir and Huelva Drifts, are evidence of a tectonic control. In the western Guadalquivir Drift, the onset of the sheeted drift construction (aggrading QII unit) above a mounded drift (prograding QI unit) resulted from a new Lower Mediterranean Core Water hydrodynamic regime. This change is correlated with a tectonic event coeval with the Mid Pleistocene Revolution (MPR) discontinuity that produced new irregularities of the seafloor during the Mid- to Late-Pleistocene. Changes in the Huelva Drift from a mounded to a sheeted drift geometry during the Late-Pleistocene, and from a prograding drift (QI and most part of QII) to an aggrading one (upper seismic unit of QII), highlight a new change in oceanographic conditions. This depositional and then oceanographic change is associated with a tectonic event, coeval with the Marine Isotope Stage (MIS) 6 discontinuity, in which a redistribution of the diapiric ridges led to the development of new local gateways, three principal branches of the Mediterranean Lower Core Water, and associated contourite channels. As a result, these buried contourite drifts hold a key palaeoceanographic record of the evolution of Mediterranean Lower Core Water, influenced by both neotectonic activity and climatic changes during the Quaternary. This study is an example of how contourite deposits and erosive elements in the marine environment can provide evidence for the reconstruction of palaeoceanographic and recent tectonic changes