Journal: The Journal of Physiology
Article Title: Loss of Baiap2l2 destabilizes the transducing stereocilia of cochlear hair cells and leads to deafness
doi: 10.1113/JP280670
Figure Lengend Snippet: A–F , current responses from OHCs of control ( Baiap2l2 tm1b/+ ) and Baiap2l2 tm1b/tm1b mice at the onset of hearing (P12: A–C ) and adult mice (P29‐40: D–F ). Currents were elicited by using depolarizing and hyperpolarizing voltage steps (10 mV increments) from the holding potential of −84 mV to the various test potentials shown by some of the traces. Note that the a large I K,n , which is carried by KCNQ4 channels, is present in OHCs at all ages and in both genotypes. Peak current–voltage curves obtained from OHCs of control and Baiap2l2 tm1b/tm1b mice are shown in ( B ) (P12) and ( E ) (P29‐40). The size of the isolated I K,n (Fig. E ) was not significantly different between the two genotypes ( C : P12: P = 0.4592; F : P29–40: P = 0.1022, t test). The number of IHCs recorded is shown next to the current–voltage traces ( B and E ) or above each column ( C and F ). For the single‐data recordings used to calculate the averages shown in ( B ) and ( E ), see Supporting information, Data S1. G , maximum intensity projections of confocal z‐stacks taken from the apical cochlear region of control and Baiap2l2 tm1b/tm1b mice at P32 using antibodies against KCNQ4 (red) and prestin (green). Phalloidin (blue) highlights the cuticular plate of the OHCs. Single cell value recordings (open symbols) are also plotted behind the average closed symbols. The number of OHCs investigated is shown above the average data points. The average OHC resting membrane potential ( V m ) and membrane capacitance ( C m ) were not significantly different between Baiap2l2 tm1b/+ ( V m −73.1 ± 1.6 mV, n = 9; C m 10.6 ± 0.6 pF, n = 8) and Baiap2l2 tm1b/tm1b mice ( V m −73.7 ± 1.3 mV, n = 6, P = 0.7528; C m 10.1 ± 0.3 pF, n = 6, P = 0.5449, t test).
Article Snippet: Primary antibodies were: rabbit‐IgG anti‐BAIAP2L2 (dilution 1:500, Atlas Antibodies, HPA003043), mouse‐IgG1 anti‐EPS8 (dilution 1:1000; 610 143; BD Biosciences, San Jose, CA, USA), mouse‐IgG1 anti‐BK channel (dilution 1:200; 75–408; NeuroMab, Davis, CA, USA), mouse‐IgG1 anti‐CtBP2 (dilution 1:50; 612 044; BD Biosciences), goat‐IgG anti‐ChAT (dilution 1:500; AB144P; Millipore, Burlington, MA, USA), rabbit‐IgG anti‐KCNQ4 (dilution 1:100; SMC‐309; StressMarq Biosciences, Victoria, BC, Canada), mouse‐IgG1 anti‐MYO7a (dilution 1:100; #138‐1s; Developmental Studies Hybridoma Bank, Iowa City, IA, USA), rabbit‐IgG anti‐MYO7a (dilution 1:500; 25–6790; Proteus Biosciences, Ramona, CA, USA) mouse‐IgG2a anti‐PSD95 (dilution 1:1000; MABN68; Millipore), rabbit‐IgG anti‐prestin (dilution 1:1000; kindly provided by Robert Fettiplace, University of Wisconsin‐Madison, USA) and rabbit‐IgG anti‐SK2 (dilution 1:500; P0483; Sigma‐Aldrich, St Louis, MO, USA).
Techniques: Isolation
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