Journal: Developmental biology

Article Title: A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea.

doi: 10.1016/j.ydbio.2011.03.031

Figure Lengend Snippet: Fig. 2. DNER and PTPζ immunolabeling of the chick's cochlea. A–C: In a wholemount preparation of the cochlear apex, DNER (red) is expressed in a punctate pattern at the apical surfaces of hair cells on the edges adjacent to the kinocilia. Phalloidin (cyan) labels the hair bundles. D–F: DNER expression (red) is strikingly lower in basal hair cells. G–I: DNER (red) is excluded from supporting cells (yellow), whose narrow apical surfaces are marked by the expression of supporting-cell antigen. J–L: The expression of DNER (red) is apparent in a section through an E8 chick vestibule. Nascent hair cells are marked by the expression by parvalbumin 3 (cyan). M–O: The expression pattern of PTPζ (cyan) is complementary to that of DNER (red). In these and subsequent illustrations of wholemounts, kinocilia are oriented toward the right. The scale bars represent 10 μm.

Article Snippet: The polyclonal antiserum against DNER (AF2254, 1 μg/μl, R and D Systems) was used at a dilution of 1:200.

Techniques: Immunolabeling, Expressing

Journal: Developmental biology

Article Title: A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea.

doi: 10.1016/j.ydbio.2011.03.031

Figure Lengend Snippet: Fig. 3. Reducing DNER expression with a splice-blocking morpholino. A–C: DNER (red) is apparent at E8 in a cryosection of nascent vestibular hair cells from an uninjected ear. Parvalbumin 3 (cyan) marks the hair-cell somata. D–F: Morpholino injection reduces the amount of DNER (red). G–I: An ear injected with control morpholino retains a regular arrangement of phalloidin-labeled hair bundles (cyan). J–L: An ear injected previously with DNER morpholino displays a patchy reduction in the concentration of DNER (red) and misorientation of hair bundles (arrowhead) labeled with phalloidin (cyan). M–O: A higher-magnification micrograph shows a central hair cell with little DNER (red) and a split hair bundle (arrowhead) labeled with phalloidin (cyan). The micrographs in G–O are surface views of E16 wholemount preparations. The scale bars represent 10 μm.

Article Snippet: The polyclonal antiserum against DNER (AF2254, 1 μg/μl, R and D Systems) was used at a dilution of 1:200.

Techniques: Expressing, Blocking Assay, Injection, Control, Labeling, Concentration Assay

Journal: Developmental biology

Article Title: A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea.

doi: 10.1016/j.ydbio.2011.03.031

Figure Lengend Snippet: Fig. 4. The effect of DNER morpholino on hair bundles. A: A scanning electron micrograph portrays two hair bundles with split clusters of stereocilia mislocalized to the edges of the apical surfaces opposite their kinocilia (arrowheads). B: A micrograph of hair bundles from near the cochlear base includes a single bundle (arrowhead) that is conspicuously taller and narrower than its neighbors. The aberrant bundle resembles those from a more apical position in the basilar papilla. The scale bars represent 2 μm.

Article Snippet: The polyclonal antiserum against DNER (AF2254, 1 μg/μl, R and D Systems) was used at a dilution of 1:200.

Techniques:

Journal: Developmental biology

Article Title: A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea.

doi: 10.1016/j.ydbio.2011.03.031

Figure Lengend Snippet: Fig. 5. Hair-bundle misorientation in DNER- and celsr1-depleted cochleas. A: In a normal E17 cochlea, a line drawn across each phalloidin-stained hair bundle (cyan) indicates its orientation. The deviation in orientations is relatively small. B: A histogram of the angular distribution of 254 hair-bundle orientations from uninjected cochleas shows a narrow peak. C: The histogram for 187 hair bundles from cochleas injected with a control morpholino is also narrow. D: In a cochlea treated with DNER morpholino, the orientations of hair bundles are disrupted. E: The angular-distribution histogram for 556 hair bundles from cochleas injected with DNER morpholino shows a broadened peak. F: Cochlear injection of a morpholino against celsr1 has a similar effect on 293 hair bundles. The scale bar represents 10 μm in panels A and D.

Article Snippet: The polyclonal antiserum against DNER (AF2254, 1 μg/μl, R and D Systems) was used at a dilution of 1:200.

Techniques: Staining, Injection, Control

Journal: Developmental biology

Article Title: A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea.

doi: 10.1016/j.ydbio.2011.03.031

Figure Lengend Snippet: Fig. 6. Misorientation of hair bundles in morpholino-injected cochleas. A–C: Injection of a morpholino against PTPζ causes disruption in the orientation pattern of hair bundles labeled with phalloidin (cyan). The localization of DNER (red) is also perturbed. D–F: A positive-control morpholino against celsr1 produces misorientation of phalloidin-labeled hair bundles (cyan) and mislocalization of DNER labeling (red). The scale bar represents 10 μm.

Article Snippet: The polyclonal antiserum against DNER (AF2254, 1 μg/μl, R and D Systems) was used at a dilution of 1:200.

Techniques: Injection, Disruption, Labeling, Positive Control

Journal: Developmental biology

Article Title: A search for factors specifying tonotopy implicates DNER in hair-cell development in the chick's cochlea.

doi: 10.1016/j.ydbio.2011.03.031

Figure Lengend Snippet: Fig. 7. The effect of DNER overexpression on hair-bundle morphology. A–D: E16 cochleas subjected to the RCAS-DNER virus at E5 display viral gag protein (yellow) both in circular hair cells and in hourglass-shaped supporting cells. In areas of viral infection, DNER (red) is overexpressed and phalloidin-labeled hair bundles (cyan) are misoriented. E–L: Higher- power micrographs with the same color coding demonstrate abnormalities of selected hair bundles marked by arrowheads. The scale bars represent 10 μm.

Article Snippet: The polyclonal antiserum against DNER (AF2254, 1 μg/μl, R and D Systems) was used at a dilution of 1:200.

Techniques: Over Expression, Virus, Infection, Labeling