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The Ca 2+ -activated Cl − channel TMEM16B regulates action potential firing and axonal targeting in olfactory sensory neurons

View Article: PubMed Central - HTML - PubMed

ABSTRACT

TMEM16B is expressed in olfactory sensory neurons, but previous attempts to establish a physiological role in olfaction have been unsuccessful. Pietra et al. find that genetic ablation of TMEM16B results in defects in the olfactory behavior of mice and the cellular physiology of olfactory sensory neurons.

No MeSH data available.


Related in: MedlinePlus

The number of I7-GFP glomeruli increased in TMEM16B KO mice. (A) The top row shows a series of three consecutive sagittal sections, where a single I7-GFP glomerulus (pointed by the arrows) is present in a WT mouse. The bottom row shows a series of consecutive sections from a KO mouse, where several glomeruli were observed (arrows). (B) Bar plot representing the total number of glomeruli per animal that significantly increased in the KO (8 ± 1, seven mice) compared with WT (4.8 ± 0.5, seven mice); unpaired t test: *, P = 0.015. (C) Bar plot representing the number of heterogeneous glomeruli in WT (black bar, 3.4 ± 3, seven mice) and in KO mice (blue bar, 5 ± 1.4, seven mice). Data are shown as mean ± SEM.
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fig9: The number of I7-GFP glomeruli increased in TMEM16B KO mice. (A) The top row shows a series of three consecutive sagittal sections, where a single I7-GFP glomerulus (pointed by the arrows) is present in a WT mouse. The bottom row shows a series of consecutive sections from a KO mouse, where several glomeruli were observed (arrows). (B) Bar plot representing the total number of glomeruli per animal that significantly increased in the KO (8 ± 1, seven mice) compared with WT (4.8 ± 0.5, seven mice); unpaired t test: *, P = 0.015. (C) Bar plot representing the number of heterogeneous glomeruli in WT (black bar, 3.4 ± 3, seven mice) and in KO mice (blue bar, 5 ± 1.4, seven mice). Data are shown as mean ± SEM.

Mentions: Mice (2–4 mo old) were sacrificed by cervical dislocation and decapitated, the head was immediately transferred to ice-cold ACSF solution, and the OBs were extracted. The fresh tissue was left overnight in a solution of 4% of paraformaldehyde (PFA) diluted in PBS. After the fixation, it was placed in 30% (wt/vol) sucrose and left overnight at 4°C for cryoprotection. 16-µm-thick sagittal sections were cut on a cryostat and stored at −20°C. Tissue sections washed in 0.1% (vol/vol) Tween 20 in PBS 30 min and rinsed with water. OB sections were incubated for 30 min with 4′-6-Diamidino-2-phenylindole (DAPI; 0.1 µg/ml) to reveal cell nuclei. Tissue sections were then washed and mounted with Vectashield (Vector Laboratories). Images were taken with an SP2 confocal microscope (Leica Microsystems) at a resolution of 1,024 × 1,024 pixels. Contrast and brightness of the images in Fig. 9 were manipulated with ImageJ software (National Institutes of Health) for the purpose of display only. Sectioning, staining, and glomerular counting were performed by an experimenter who was blinded to the genotype of the mice.


The Ca 2+ -activated Cl − channel TMEM16B regulates action potential firing and axonal targeting in olfactory sensory neurons
The number of I7-GFP glomeruli increased in TMEM16B KO mice. (A) The top row shows a series of three consecutive sagittal sections, where a single I7-GFP glomerulus (pointed by the arrows) is present in a WT mouse. The bottom row shows a series of consecutive sections from a KO mouse, where several glomeruli were observed (arrows). (B) Bar plot representing the total number of glomeruli per animal that significantly increased in the KO (8 ± 1, seven mice) compared with WT (4.8 ± 0.5, seven mice); unpaired t test: *, P = 0.015. (C) Bar plot representing the number of heterogeneous glomeruli in WT (black bar, 3.4 ± 3, seven mice) and in KO mice (blue bar, 5 ± 1.4, seven mice). Data are shown as mean ± SEM.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5037344&req=5

fig9: The number of I7-GFP glomeruli increased in TMEM16B KO mice. (A) The top row shows a series of three consecutive sagittal sections, where a single I7-GFP glomerulus (pointed by the arrows) is present in a WT mouse. The bottom row shows a series of consecutive sections from a KO mouse, where several glomeruli were observed (arrows). (B) Bar plot representing the total number of glomeruli per animal that significantly increased in the KO (8 ± 1, seven mice) compared with WT (4.8 ± 0.5, seven mice); unpaired t test: *, P = 0.015. (C) Bar plot representing the number of heterogeneous glomeruli in WT (black bar, 3.4 ± 3, seven mice) and in KO mice (blue bar, 5 ± 1.4, seven mice). Data are shown as mean ± SEM.
Mentions: Mice (2–4 mo old) were sacrificed by cervical dislocation and decapitated, the head was immediately transferred to ice-cold ACSF solution, and the OBs were extracted. The fresh tissue was left overnight in a solution of 4% of paraformaldehyde (PFA) diluted in PBS. After the fixation, it was placed in 30% (wt/vol) sucrose and left overnight at 4°C for cryoprotection. 16-µm-thick sagittal sections were cut on a cryostat and stored at −20°C. Tissue sections washed in 0.1% (vol/vol) Tween 20 in PBS 30 min and rinsed with water. OB sections were incubated for 30 min with 4′-6-Diamidino-2-phenylindole (DAPI; 0.1 µg/ml) to reveal cell nuclei. Tissue sections were then washed and mounted with Vectashield (Vector Laboratories). Images were taken with an SP2 confocal microscope (Leica Microsystems) at a resolution of 1,024 × 1,024 pixels. Contrast and brightness of the images in Fig. 9 were manipulated with ImageJ software (National Institutes of Health) for the purpose of display only. Sectioning, staining, and glomerular counting were performed by an experimenter who was blinded to the genotype of the mice.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

TMEM16B is expressed in olfactory sensory neurons, but previous attempts to establish a physiological role in olfaction have been unsuccessful. Pietra et al. find that genetic ablation of TMEM16B results in defects in the olfactory behavior of mice and the cellular physiology of olfactory sensory neurons.

No MeSH data available.


Related in: MedlinePlus