Syntaxin 4 is essential for hearing in human and zebrafish.

Congenital hearing impairment (HI) is a genetically highly heterogeneous disorder in which prompt recognition and intervention are crucial to optimize outcomes. In this study, we used exome sequencing to investigate a large consanguineous Pakistani family with eight affected individuals showing bilateral severe-to-profound HI. This identified a homozygous splice region variant in STX4 (c.232 + 6T>C), which causes exon skipping and a frameshift, that segregated with HI (two-point logarithm of odds (LOD) score = 5.9). STX4, a member of the syntaxin family, is a component of the SNARE machinery involved in several vesicle transport and recycling pathways. In silico analysis showed that murine orthologue Stx4a is highly and widespread expressed in the developing and adult inner ear. Immunofluorescent imaging revealed localization of STX4A in the cell body, cell membrane and stereocilia of inner and outer hair cells. Furthermore, a morpholino-based knockdown of stx4 in zebrafish showed an abnormal startle response, morphological and developmental defects, and a disrupted mechanotransduction function in neuromast hair cells measured via FM1-43 uptake. Our findings indicate that STX4 dysfunction leads to HI in humans and zebrafish and supports the evolutionary conserved role of STX4 in inner ear development and hair cell functioning.

A novel autosomal dominant GREB1L variant associated with non-syndromic hearing impairment in Ghana.

BACKGROUND: Childhood hearing impairment (HI) is genetically heterogeneous with many implicated genes, however, only a few of these genes are reported in African populations. METHODS: This study used exome and Sanger sequencing to resolve the possible genetic cause of non-syndromic HI in a Ghanaian family. RESULTS: We identified a novel variant c.3041G > A: p.(Gly1014Glu) in GREB1L (DFNA80) in the index case. The GREB1L: p.(Gly1014Glu) variant had a CADD score of 26.5 and was absent from human genomic databases such as TopMed and gnomAD. In silico homology protein modeling approaches displayed major structural differences between the wildtype and mutant proteins. Additionally, the variant was predicted to probably affect the secondary protein structure that may impact its function. Publicly available expression data shows a higher expression of Greb1L in the inner ear of mice during development and a reduced expression in adulthood, underscoring its importance in the development of the inner ear structures. CONCLUSION: This report on an African individual supports the association of GREB1L variant with non-syndromic HI and extended the evidence of the implication of GREB1L variants in HI in diverse populations.

Rapid broadband discrete nanomechanical mapping of soft samples on atomic force microscope.

In this paper, an approach to achieve rapid broadband discrete nanomechanical mapping of soft samples using an atomic force microscope is developed. Nanomechanical mapping (NM) is needed to investigate, for example, dynamic evolution of the nanomechanical distribution of the sample-provided that the mapping is fast enough. The throughput of conventional NM methods, however, is inherently limited by the continuous scanning involved where the probe visits each sampling location continuously. Thus, we propose to significantly reduce the number of measurements through discrete mapping where only discrete sampling locations of interests are visited and measured. An online-searching learning-based technique is utilized to achieve rapid probe engagement and withdrawal with the interaction force minimized at each sampling location. Then, a control-based nanoindentation measurement technique is used to quickly acquire the nanomechanical property at each location, over frequencies that can be chosen arbitrarily in a broad range. Finally, a decomposition-based learning approach is explored to achieve rapid probe transitions between the sampling locations. The proposed technique is demonstrated through experiments using a Polydimethylsiloxane (PDMS) sample and a PDMS-epoxy sample as examples.