DNALI1 deficiency causes male infertility with severe asthenozoospermia in humans and mice by disrupting the assembly of the flagellar inner dynein arms and fibrous sheath

Abstract

The axonemal dynein arms (outer (ODA) and inner dynein arms (IDAs)) are multiprotein structures organized by light, intermediate, light intermediate (LIC), and heavy chain proteins. They hydrolyze ATP to promote ciliary and flagellar movement. Till now, a variety of dynein protein deficiencies have been linked with asthenospermia (ASZ), highlighting the significance of these structures in human sperm motility. Herein, we detected bi-allelic DNALI1 mutations [c.663_666del (p.Glu221fs)], in an ASZ patient, which resulted in the complete loss of the DNALI1 in the patient’s sperm. We identified loss of sperm DNAH1 and DNAH7 rather than DNAH10 in both DNALI1663_666del patient and Dnali1−/− mice, demonstrating that mammalian DNALI1 is a LIC protein of a partial IDA subspecies. More importantly, we revealed that DNALI1 loss contributed to asymmetries in the most fibrous sheath (FS) of the sperm flagellum in both species. Immunoprecipitation revealed that DNALI1 might interact with the cytoplasmic dynein complex proteins in the testes. Furthermore, DNALI1 loss severely disrupted the transport and assembly of the FS proteins, especially AKAP3 and AKAP4, during flagellogenesis. Hence, DNALI1 may possess a non-classical molecular function, whereby it regulates the cytoplasmic dynein complex that assembles the flagella. We conclude that a DNALI deficiency-induced IDAs injury and an asymmetric FS-driven tail rigid structure alteration may simultaneously cause flagellum immotility. Finally, intracytoplasmic sperm injection (ICSI) can effectively resolve patient infertility. Collectively, we demonstrate that DNALI1 is a newly causative gene for AZS in both humans and mice, which possesses multiple crucial roles in modulating flagellar assembly and motility.

AXDND1 is required to balance spermatogonial commitment and for sperm tail formation in mice and humans

“…Therefore, AXDND1 may play dual roles in dynein complexes to accomplish axoneme-mediated cell motility and for cytoplasmic dynein-mediated transport. A similar hypothesis has been raised for the light intermediate dynein chain DNALI1 (32). A variant encoding hypomorphic DNALI1 in mice resulted in axoneme microtubule doublet and fibrous sheath abnormalities (32).…”

“…A similar hypothesis has been raised for the light intermediate dynein chain DNALI1 (32). A variant encoding hypomorphic DNALI1 in mice resulted in axoneme microtubule doublet and fibrous sheath abnormalities (32). Alternatively, the motility defect observed in sperm from Axdnd1 -/- males may be exclusively due to the severe ultrastructural defects, including the partial loss of microtubule doublets and outer dense fibres throughout the axoneme.…”

Novel biallelic variants in DNAH1 cause multiple morphological abnormalities of sperm flagella with favorable outcomes of fertility after ICSI in Han Chinese males

“…As previously reported, DNAH1 deficiency impaired the structure of IDAs and induced morphological abnormalities of spermatozoa in infertile humans and mice 9,12 . To explore the effect of DNAH1 variants on the molecular characteristics of spermatozoa, immunofluorescence assays with antibodies against DNAH1, DNALI1 (a light intermediate chain protein of IDA), 36 DNAI1 (an intermediate chain protein of ODA), 37 and α‐TUBULIN (the main component of microtubules) were performed. The results showed that biallelic DNAH1 variants induced the absence of DNAH1 and a decrease in α‐TUBULIN in the spermatozoa of males with MMAF (Figure 4A).…”

SWATH-MS Analysis of Blood Plasma and Circulating Small Extracellular Vesicles Enables Detection of Putative Protein Biomarkers of Fertility in Young and Aged Dairy Cows

“…For instance, Dna14, which encodes Dynein axonemal intermediate zoa motility and ciliary transport of the oocyte in the oviduct and fallopian tube is the most relevant. 41,42 As the specific function of DNAI4 has not yet been described in this respect, future investigations may wish to elucidate the role of this specific dynein in in vitro or in vivo models. The other uncharacterized protein in the high-fertility sEV group, A5PKB9 (C16orf54) has been implicated as a potential diagnostic/prognostic in Pancancer, 43 and in lipid droplet formation, 44 thus its association with reproductive status is unclear.…”

“…Interestingly, each high- and low-fertility protein biomarker group contained two proteins that have not been previously classified as vesicular in origin in the online EV database Vesiclepedia and may represent a novel finding. For instance, Dna14 , which encodes Dynein axonemal intermediate chain 4 (DNAI4), was exclusively detected in high-fertility sEV samples and has not been well-characterized; however, other members of the dynein complex family have been under study for their roles in vertebrate male and female infertility, namely dynein axenomal intermediate and light chains DNAI1 and DNAL1. − The dynein family of motor proteins perform a wide range of biological functions, but in the context of reproduction and reproductive deficits, their role in spermatozoa motility and ciliary transport of the oocyte in the oviduct and fallopian tube is the most relevant. , As the specific function of DNAI4 has not yet been described in this respect, future investigations may wish to elucidate the role of this specific dynein in in vitro or in vivo models. The other uncharacterized protein in the high-fertility sEV group, A5PKB9 ( C16orf54 ) has been implicated as a potential diagnostic/prognostic in Pan-cancer, and in lipid droplet formation, thus its association with reproductive status is unclear.…”