Journal: European Journal of Human Genetics

Article Title: ADAMTS1 , MPDZ, MVD , and SEZ6: candidate genes for autosomal recessive nonsyndromic hearing impairment

doi: 10.1038/s41431-021-00913-x

Figure Lengend Snippet: Information on the variants found in the four candidate NSHI genes.

Article Snippet: The following primary antibodies were used: sheep anti-mouse ADAMTS1 (AF5867-SP; Novus Biologicals, Centennial, CO, USA), rabbit anti-mouse MPDZ (42-2700; Thermofisher Scientific, Waltham, MA, USA), rabbit anti-mouse MVD (PA5-22164; Thermofisher Scientific), and sheep anti-mouse SEZ6 (PA5-47683; Thermofisher Scientific).

Techniques:

Journal: European Journal of Human Genetics

Article Title: ADAMTS1 , MPDZ, MVD , and SEZ6: candidate genes for autosomal recessive nonsyndromic hearing impairment

doi: 10.1038/s41431-021-00913-x

Figure Lengend Snippet: Panel A : Structural model of the wild type ADAMTS1, MPDZ, MVD, and SEZ6 proteins. Panel B : Interacting bond of the wild type proteins for the four candidate genes. Panel C : Mutant type proteins highlighting the change due to the variant. The variants ADAMTS1 [p.(Ser135Ala)] and MPDZ [p.(Pro775Leu)] may result in a difference in interaction pattern with the solvent molecules. As the position of the variant residue is at the loop region, they do not involve a direct interaction, but the native fold of the protein may be affected. MVD [p.Pro379His] mutated protein shows a difference in interacting bond distance in the mutant type from the wild type and may perturb the amino acid side chain. The SEZ6 [p.Val698Ile] mutant protein displayed less interaction distance due to substitution of Val to Ile at position 698, while the SEZ6 wild type protein has a weak H-bond and hydrophobic interaction with Ile635 and Trp609. This variation could possibly change the orientation of the amino acid residue. Structures are displayed as ribbon while residue is represented by stick model. The structure illustrations were created with the PyMOL program. Black dotted lines represent hydrogen bonds.

Article Snippet: The following primary antibodies were used: sheep anti-mouse ADAMTS1 (AF5867-SP; Novus Biologicals, Centennial, CO, USA), rabbit anti-mouse MPDZ (42-2700; Thermofisher Scientific, Waltham, MA, USA), rabbit anti-mouse MVD (PA5-22164; Thermofisher Scientific), and sheep anti-mouse SEZ6 (PA5-47683; Thermofisher Scientific).

Techniques: Mutagenesis, Variant Assay, Solvent, Residue

Journal: European Journal of Human Genetics

Article Title: ADAMTS1 , MPDZ, MVD , and SEZ6: candidate genes for autosomal recessive nonsyndromic hearing impairment

doi: 10.1038/s41431-021-00913-x

Figure Lengend Snippet: A RNA expression of the four candidate genes in the cochlea and utricle during mouse development. RNA-sequencing data of hair cells (GFP+) and surrounding cells (GFP−) from the cochleae and utricles of mice expressing EGFP under the Pou4f3 promoter. Data were collected at four developmental stages: E16, P0, P4, and P7 and RNA expression is represented as normalized counts. Adamts1 , Mpdz , Mvd , and Sez6 were expressed in the hair cells and surrounding cells of the cochlea and utricle during the E16, P0, P4, and P7 stages. B RNA expression of the four candidate genes in the adult inner ear cells. The figure represents expression of the genes of interest in Deiters’ cells (Deiters), Pillar cells (Pillar), Inner Hair Cells (IHC), and Outer Hair Cells (OHC) of adult CBA/J mice. The four candidate genes Adamts1, Mpdz, Mvd , and Sez6 were expressed in the IHC, OHCs, and Deiter’s and Pillar cells. The y-axis represents the gene expression normalized to transcripts per million (TPM). The dataset was obtained from the GEO database (accession number GSE111347).

Article Snippet: The following primary antibodies were used: sheep anti-mouse ADAMTS1 (AF5867-SP; Novus Biologicals, Centennial, CO, USA), rabbit anti-mouse MPDZ (42-2700; Thermofisher Scientific, Waltham, MA, USA), rabbit anti-mouse MVD (PA5-22164; Thermofisher Scientific), and sheep anti-mouse SEZ6 (PA5-47683; Thermofisher Scientific).

Techniques: RNA Expression, RNA Sequencing, Expressing, Gene Expression

Journal: European Journal of Human Genetics

Article Title: ADAMTS1 , MPDZ, MVD , and SEZ6: candidate genes for autosomal recessive nonsyndromic hearing impairment

doi: 10.1038/s41431-021-00913-x

Figure Lengend Snippet: A Whole mount immunostaining of Adamts1, Mpdz, and Sez6 in wildtype mice at P12. Immunoreactivity was visualized with a fluorescently labeled secondary antibody (red) and F-actin was stained with phalloidin 488 nm (green). Immunolabeling of Adamts1 and Sez6 is observed in stereocilia as well as cytoplasm of outer hair cells. Mpdz is observed in the cytoplasmic region of hair cells. B Immunostaining of the organ of Corti at the apical, medial and basal turns of the cochlea at P4. C Immunostaining of Mvd in wildtype mice at P1, P4, and P28. Immunoreactivity of Mvd was visualized with a fluorescently labeled secondary antibody (green), F-actin was stained with rhodamine-phalloidin (red) and nuclear bodies were stained with DAPI (blue). The anti-neurofilament (NF-200, purple) was used to mark the neurons. Immunolabeling of Mvd is observed in spiral ganglion cells (SG). HP Habenula perforata, OC Organ of Corti.

Article Snippet: The following primary antibodies were used: sheep anti-mouse ADAMTS1 (AF5867-SP; Novus Biologicals, Centennial, CO, USA), rabbit anti-mouse MPDZ (42-2700; Thermofisher Scientific, Waltham, MA, USA), rabbit anti-mouse MVD (PA5-22164; Thermofisher Scientific), and sheep anti-mouse SEZ6 (PA5-47683; Thermofisher Scientific).

Techniques: Immunostaining, Labeling, Staining, Immunolabeling

Journal: European Journal of Human Genetics

Article Title: ADAMTS1 , MPDZ, MVD , and SEZ6: candidate genes for autosomal recessive nonsyndromic hearing impairment

doi: 10.1038/s41431-021-00913-x

Figure Lengend Snippet: Functions, expression and animal model phenotypes for the four NSHI candidate genes.

Article Snippet: The following primary antibodies were used: sheep anti-mouse ADAMTS1 (AF5867-SP; Novus Biologicals, Centennial, CO, USA), rabbit anti-mouse MPDZ (42-2700; Thermofisher Scientific, Waltham, MA, USA), rabbit anti-mouse MVD (PA5-22164; Thermofisher Scientific), and sheep anti-mouse SEZ6 (PA5-47683; Thermofisher Scientific).

Techniques: Expressing, Animal Model, Protein-Protein interactions, Membrane

Journal: Journal of Neurochemistry

Article Title: Astrocyte‐specific transcriptome analysis using the ALDH1L1 bacTRAP mouse reveals novel biomarkers of astrogliosis in response to neurotoxicity

doi: 10.1111/jnc.14800

Figure Lengend Snippet: Gene ontology categorization of DEGs identified in astrocytes post‐1‐methyl‐4‐phenyl‐1,2,3,6,‐tetrahydropyridine (MPTP) exposure

Article Snippet: Real‐time PCR analysis of the housekeeping gene, GAPDH (Cat# 4331182, Mm99999915_g1/Rn01775763_g1), and target genes: TIMP1 (Cat# 4331182, Mm01341361_m1/Rn01430873_g1), SERPINF2 (Cat# 4331182, Mm00435868_m1/Rn01464595_m1), AA467197/miR‐147 (Cat# 4351372, Mm01268692_m1), SOCS3 (Cat# 4331182, Mm00545913_s1/Rn00585674_s1), ADAMTS1 (Cat# 4351372, Mm01244169_m1/Cat# 4331182, Rn00577887_m1), GLDN (Cat# 4331182, Mm00616548_m1/Rn00710589_m1), NES (Cat# 4331182, Mm00450205_m1/ Cat# 4351372, Rn01455599_g1), ECM1 (Cat# 4331182, Mm00514634_m1/Rn01534170_g1), and FXYD5 (Cat# 4331182, Mm00435435_m1/Rn00573226_m1) was performed using FAM‐ MGB TaqMan® primers from Thermo Fisher and an ABI7500 Real‐Time PCR System (Thermo Fisher Scientific) in combination with TaqMan® chemistry as previously described (Locker et al. , ; Kelly et al. , ).

Techniques: Binding Assay, Activity Assay, Protein Binding