Journal: Frontiers in Neuroscience

Article Title: Dystrophin Is Required for the Proper Timing in Retinal Histogenesis: A Thorough Investigation on the mdx Mouse Model of Duchenne Muscular Dystrophy

doi: 10.3389/fnins.2020.00760

Figure Lengend Snippet: Primary antibodies used for immunofluorescence.

Article Snippet: Rabbit anti-calbindin (HC) , 1:1000 , Merck Millipore (Billerica, MA, United States).

Techniques: Immunofluorescence

Journal: eNeuro

Article Title: Demyelination and Na + Channel Redistribution Underlie Auditory and Vestibular Dysfunction in PMP22-Null Mice

doi: 10.1523/ENEURO.0462-23.2023

Figure Lengend Snippet: Compared with wild-type mice, reduced expression of MBP and MPZ in auditory and vestibular neurons in PMP22 −/− cochlea. A , Cross sections through the inner ear of PMP22 +/+ and PMP22 −/− showing an overview of the organ of Corti (OC), SGNs, central ANFs, modiolar (M), vestibular neurons (VNs), and vestibular neuron fibers (VNFs). MPZ is in red, and MBP is in green. Scale bar, 100 µm. B , SGNs were stained with Tuj1 (green), and the myelinated fibers were identified with MBP (red) antibodies. The top panels show images from PMP22 +/+ mice, while the bottom panels are the PMP22 −/− cochlea. The neurites are devoid of MBP, but patches of MBP labeling can be seen. Scale bar, 20 µm. C , A longitudinal section of the peripheral aspects of the auditory nerve was stained with Tuj1 (cyan), MBP (red), and MPZ (green). For orientation, the OC is indicated. Besides a few patches of MBP and MPZ expression in the PMP22 −/− auditory nerve, they are devoid of myelin proteins compared with the PMP22 +/+ . Scale bar, 20 µm. D , Sections through the ANFs from PMP22 +/+ and PMP22 −/− mice are shown. Scale bar, 5 µm. E , Sections through the VN reveal similar nerve fibers and neurons bare of myelin proteins. Scale bar, 10 µm.

Article Snippet: Microdissected pieces were immunostained with antibodies to the following: (1) mouse anti-C-terminal binding protein 2 (presynaptic marker, BD Biosciences, 1:200); (2) rabbit anti-myosin VIIa (HC marker, Proteus Biosciences, 1:600); (3) rabbit anti-Homer 1 (postsynaptic marker, Synaptic Systems, 1:250); (4) rabbit anti-Tuj1 (BioLegend, 1:500), chicken anti-Tuj1 (Abcam,1:500), and mouse anti-Tuj1 (Abcam, 1:500); (5) rabbit anti-PMP22 (Abcam, 1:500); (6) chicken anti-MBP (EnCor Biotechnology, 1:500), rabbit anti-MBP (Abcam, 1:500), mouse IgG2b anti-MBP (Abcam, 1:500); (7) chicken anti-MPZ (Abcam, 1:500); (8) goat anti-calretinin (Swant, 1:500); (9) mouse anti-calretinin (MilliporeSigma, 1:200); and (10) rabbit anti-calbindin (Cell Signaling Technology, 1:200) with appropriate secondary antibodies coupled to Alexa 405, 488, 568, and 647 fluorophores from Jackson ImmunoResearch and Thermo Fisher Scientific.

Techniques: Expressing, Staining, Labeling

Journal: eNeuro

Article Title: Demyelination and Na + Channel Redistribution Underlie Auditory and Vestibular Dysfunction in PMP22-Null Mice

doi: 10.1523/ENEURO.0462-23.2023

Figure Lengend Snippet: In PMP22 −/− vestibular neurons compared with PMP22 +/+ mice, MBP and MPZ expression is reduced. A , B , Cross section through the inner ear of PMP22 +/+ and PMP22 −/− mice showing Scarpa ganglion vestibular neurons. MPZ is in green, and MBP is in red. Tuj1 staining serves as a neuronal marker, and DAPI is used for nuclear staining. Glial cells in the PMP22 −/− mice display low levels of myelin proteins and, when expressed, appear condensed. Scale bar, 5 µm.

Article Snippet: Microdissected pieces were immunostained with antibodies to the following: (1) mouse anti-C-terminal binding protein 2 (presynaptic marker, BD Biosciences, 1:200); (2) rabbit anti-myosin VIIa (HC marker, Proteus Biosciences, 1:600); (3) rabbit anti-Homer 1 (postsynaptic marker, Synaptic Systems, 1:250); (4) rabbit anti-Tuj1 (BioLegend, 1:500), chicken anti-Tuj1 (Abcam,1:500), and mouse anti-Tuj1 (Abcam, 1:500); (5) rabbit anti-PMP22 (Abcam, 1:500); (6) chicken anti-MBP (EnCor Biotechnology, 1:500), rabbit anti-MBP (Abcam, 1:500), mouse IgG2b anti-MBP (Abcam, 1:500); (7) chicken anti-MPZ (Abcam, 1:500); (8) goat anti-calretinin (Swant, 1:500); (9) mouse anti-calretinin (MilliporeSigma, 1:200); and (10) rabbit anti-calbindin (Cell Signaling Technology, 1:200) with appropriate secondary antibodies coupled to Alexa 405, 488, 568, and 647 fluorophores from Jackson ImmunoResearch and Thermo Fisher Scientific.

Techniques: Expressing, Staining, Marker

Journal: bioRxiv

Article Title: Genetic and pharmacologic alterations of claudin9 levels suffice to induce functional and mature inner hair cells

doi: 10.1101/2023.10.08.561387

Figure Lengend Snippet: A. Quantitative RT-PCR of Cldn9 transcripts from cochlear tissue from six groups of animals, including Cldn9 T/T , Cldn9 +/T with and without dox treatment compared with WT littermates with and without dox treatment. B. Body weight measurements of female and male mice from dox-treated Cldn9 +/T and Cldn9 +/+ groups were recorded at 4, 6, and 8 wks old. C. The immunostaining of Cldn9 in 8-wk old WT ( Cldn9 +/+ ) mouse cochlea Cldn9 (red), IHC stained myosin7a (cyan) and supporting cells stained Sox2 (green) and Dapi (blue) for nuclear stain. Scale = 10 μm.

Article Snippet: Microdissected pieces were immunostained with antibodies to the following: (1) mouse anti-C-terminal binding protein 2 (pre-synaptic-marker, BD Biosciences, 1:200, Cat # 612044), (2) rabbit anti-myosin-VIIa (HC-marker, Proteus Biosciences, Inc,1:600, Cat # 25-6790), (3) mouse anti-sox2 (supporting cell (SC)-marker, Santa Cruz Biotechnology, Inc, 1:200, Cat # sc-365823), and (4) rabbit anti-Homer 1 (post-synaptic marker, Synaptic Systems, 1:250, Cat # 160 003), (5) rabbit anti-immunoglobulin like domain containing receptor 1 (ILDR1) (Antibodies-online.com, 1:200, Cat # ABIN1386369), (6) mouse anti-Cldn9 (Santa Cruz, 1:200, Cat # sc-398836), (7) mouse anti-Cldn6 (Santa Cruz, 1:200, Cat # sc-393671), (8) rabbit anti-Sox2 (Abcam, 1:200, Cat # ab97959), (9) rabbit anti-Tuj1 and chicken anti-Tuj1 (Abcam,1:500, Cat # ab18207, ab41489), (10) goat anti-calretinin (Swant Inc., 1:500, code # CG1), (11) mouse anti-calretinin (Millipore Sigma, 1:200, Cat # MAB1568), (12) rabbit anti-Prestin (Santa Cruz, 1:200, Cat # sc-22692), and (13) rabbit anti-calbindin (Cell signaling technology, 1:200, Cat # 13176S) with appropriate secondary antibodies coupled to Alexa-405, −488, −568, and −647 fluorophores.. DAPI labeled the cell nucleus after secondary antibody incubation.

Techniques: Quantitative RT-PCR, Immunostaining, Staining

Journal: bioRxiv

Article Title: Genetic and pharmacologic alterations of claudin9 levels suffice to induce functional and mature inner hair cells

doi: 10.1101/2023.10.08.561387

Figure Lengend Snippet: A-B , The immunostaining of Cldn6 (red) in the mouse cochlea from Cldn9 +/+ (wildtype, WT) and C ldn9 +/T . The lower Panel is the side view of the cochlear section. HCs were labeled with Myo7a (cyan) supporting cells with Sox2 (green) and Dapi-stained (blue) nuclei. The levels of Cldn6 were reduced in the Cldn9 +/T cochlea. C-D , The immunostaining of ILDR1 (red) in the mouse cochlea from Cldn9 +/+ and C ldn9 +/T . The lower Panel is the side view of the cochlear section. HCs were labeled with Myo7a (cyan) supporting cells with Sox2 (green) and Dapi-stained (blue) nuclei. The levels of ILDR1 were increased in the Cldn9 +/T cochlea. Scale bar = 10 μm.

Article Snippet: Microdissected pieces were immunostained with antibodies to the following: (1) mouse anti-C-terminal binding protein 2 (pre-synaptic-marker, BD Biosciences, 1:200, Cat # 612044), (2) rabbit anti-myosin-VIIa (HC-marker, Proteus Biosciences, Inc,1:600, Cat # 25-6790), (3) mouse anti-sox2 (supporting cell (SC)-marker, Santa Cruz Biotechnology, Inc, 1:200, Cat # sc-365823), and (4) rabbit anti-Homer 1 (post-synaptic marker, Synaptic Systems, 1:250, Cat # 160 003), (5) rabbit anti-immunoglobulin like domain containing receptor 1 (ILDR1) (Antibodies-online.com, 1:200, Cat # ABIN1386369), (6) mouse anti-Cldn9 (Santa Cruz, 1:200, Cat # sc-398836), (7) mouse anti-Cldn6 (Santa Cruz, 1:200, Cat # sc-393671), (8) rabbit anti-Sox2 (Abcam, 1:200, Cat # ab97959), (9) rabbit anti-Tuj1 and chicken anti-Tuj1 (Abcam,1:500, Cat # ab18207, ab41489), (10) goat anti-calretinin (Swant Inc., 1:500, code # CG1), (11) mouse anti-calretinin (Millipore Sigma, 1:200, Cat # MAB1568), (12) rabbit anti-Prestin (Santa Cruz, 1:200, Cat # sc-22692), and (13) rabbit anti-calbindin (Cell signaling technology, 1:200, Cat # 13176S) with appropriate secondary antibodies coupled to Alexa-405, −488, −568, and −647 fluorophores.. DAPI labeled the cell nucleus after secondary antibody incubation.

Techniques: Immunostaining, Labeling, Staining