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Development of the Olfactory Epithelium and Nasal Glands in TMEM16A-/- and TMEM16A+/+ Mice.

Maurya DK, Henriques T, Marini M, Pedemonte N, Galietta LJ, Rock JR, Harfe BD, Menini A - PLoS ONE (2015)

Bottom Line: A comparison between the expression of the olfactory marker protein and adenylyl cyclase III shows that genetic ablation of TMEM16A did not seem to affect the maturation of olfactory sensory neurons and their ciliary layer.Moreover, we also observed that the morphology of Bowman's glands, nasal septal glands and lateral nasal glands did not change in the absence of TMEM16A.Our results indicate that the development of mouse olfactory epithelium and nasal glands does not seem to be affected by the genetic ablation of TMEM16A.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Olfactory Transduction, SISSA, International School for Advanced Studies, Trieste, Italy.

ABSTRACT
TMEM16A/ANO1 is a calcium-activated chloride channel expressed in several types of epithelia and involved in various physiological processes, including proliferation and development. During mouse embryonic development, the expression of TMEM16A in the olfactory epithelium is dynamic. TMEM16A is expressed at the apical surface of the entire olfactory epithelium at embryonic day E12.5 while from E16.5 its expression is restricted to a region near the transition zone with the respiratory epithelium. To investigate whether TMEM16A plays a role in the development of the mouse olfactory epithelium, we obtained the first immunohistochemistry study comparing the morphological properties of the olfactory epithelium and nasal glands in TMEM16A-/- and TMEM16A+/+ littermate mice. A comparison between the expression of the olfactory marker protein and adenylyl cyclase III shows that genetic ablation of TMEM16A did not seem to affect the maturation of olfactory sensory neurons and their ciliary layer. As TMEM16A is expressed at the apical part of supporting cells and in their microvilli, we used ezrin and cytokeratin 8 as markers of microvilli and cell body of supporting cells, respectively, and found that morphology and development of supporting cells were similar in TMEM16A-/- and TMEM16A+/+ littermate mice. The average number of supporting cells, olfactory sensory neurons, horizontal and globose basal cells were not significantly different in the two types of mice. Moreover, we also observed that the morphology of Bowman's glands, nasal septal glands and lateral nasal glands did not change in the absence of TMEM16A. Our results indicate that the development of mouse olfactory epithelium and nasal glands does not seem to be affected by the genetic ablation of TMEM16A.

No MeSH data available.


Related in: MedlinePlus

Expression of TMEM16A in various nasal glands of TMEM16A-/- and TMEM16A+/+ littermate mice.Expression of TMEM16A and aquaporin 5 in the Bowman’s gland (BG), nasal septal gland (NSG) and lateral nasal gland (LNG) of TMEM16A-/- and TMEM16A+/+ littermate mice. A, D: aquaporin 5 immunopositive signals were seen in Bowman’s glands in P4 mice. Aquaporin 5 expression in glands and ducts is clearly visible. TMEM16A immunoreactivity (goat anti-TMEM16A) was not present in Bowman’s glands of TMEM16A-/- nor of TMEM16A+/+ littermate mice (A, D). However, aquaporin 5 and TMEM16A were co-expressed in nasal septal glands (B) and lateral nasal glands (C) of TMEM16A+/+ mice. No immunoreactivity to TMEM16A was detectable in TMEM16A-/- mice (E, F). Glands marked by aquaporin 5 were similar in both types of mice. Images are averages of z-stacks of thickness of ~2.0 μm for A, D, or ~1 μm for B, C, E, F. Cell nuclei were stained by DAPI. Scale bars = 10 μm.
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pone.0129171.g008: Expression of TMEM16A in various nasal glands of TMEM16A-/- and TMEM16A+/+ littermate mice.Expression of TMEM16A and aquaporin 5 in the Bowman’s gland (BG), nasal septal gland (NSG) and lateral nasal gland (LNG) of TMEM16A-/- and TMEM16A+/+ littermate mice. A, D: aquaporin 5 immunopositive signals were seen in Bowman’s glands in P4 mice. Aquaporin 5 expression in glands and ducts is clearly visible. TMEM16A immunoreactivity (goat anti-TMEM16A) was not present in Bowman’s glands of TMEM16A-/- nor of TMEM16A+/+ littermate mice (A, D). However, aquaporin 5 and TMEM16A were co-expressed in nasal septal glands (B) and lateral nasal glands (C) of TMEM16A+/+ mice. No immunoreactivity to TMEM16A was detectable in TMEM16A-/- mice (E, F). Glands marked by aquaporin 5 were similar in both types of mice. Images are averages of z-stacks of thickness of ~2.0 μm for A, D, or ~1 μm for B, C, E, F. Cell nuclei were stained by DAPI. Scale bars = 10 μm.

Mentions: Fig 8A and 8D shows that aquaporin 5 stained the internal wall of the duct of a Bowman’s gland as well as microvilli of supporting cells. Previous reports showed expression of TMEM16A in the duct of Bowman’s glands and in the luminal surface of nasal septal glands and lateral nasal glands [16]. However, in TMEM16A+/+ mice, we did not find expression of TMEM16A in Bowman’s glands, whereas the luminal surface of nasal septal glands and lateral nasal glands expressed both TMEM16A and aquaporin 5 (Fig 8B and 8C). A comparison with results from TMEM16A-/- mice shows that immunostaining of aquaporin 5 remains unchanged, whereas TMEM16A immunoreactivity was absent (Fig 8D–8F). The morphology of Bowman’s gland, nasal septal glands and lateral nasal glands remained the same in both types of mice.


Development of the Olfactory Epithelium and Nasal Glands in TMEM16A-/- and TMEM16A+/+ Mice.

Maurya DK, Henriques T, Marini M, Pedemonte N, Galietta LJ, Rock JR, Harfe BD, Menini A - PLoS ONE (2015)

Expression of TMEM16A in various nasal glands of TMEM16A-/- and TMEM16A+/+ littermate mice.Expression of TMEM16A and aquaporin 5 in the Bowman’s gland (BG), nasal septal gland (NSG) and lateral nasal gland (LNG) of TMEM16A-/- and TMEM16A+/+ littermate mice. A, D: aquaporin 5 immunopositive signals were seen in Bowman’s glands in P4 mice. Aquaporin 5 expression in glands and ducts is clearly visible. TMEM16A immunoreactivity (goat anti-TMEM16A) was not present in Bowman’s glands of TMEM16A-/- nor of TMEM16A+/+ littermate mice (A, D). However, aquaporin 5 and TMEM16A were co-expressed in nasal septal glands (B) and lateral nasal glands (C) of TMEM16A+/+ mice. No immunoreactivity to TMEM16A was detectable in TMEM16A-/- mice (E, F). Glands marked by aquaporin 5 were similar in both types of mice. Images are averages of z-stacks of thickness of ~2.0 μm for A, D, or ~1 μm for B, C, E, F. Cell nuclei were stained by DAPI. Scale bars = 10 μm.
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pone.0129171.g008: Expression of TMEM16A in various nasal glands of TMEM16A-/- and TMEM16A+/+ littermate mice.Expression of TMEM16A and aquaporin 5 in the Bowman’s gland (BG), nasal septal gland (NSG) and lateral nasal gland (LNG) of TMEM16A-/- and TMEM16A+/+ littermate mice. A, D: aquaporin 5 immunopositive signals were seen in Bowman’s glands in P4 mice. Aquaporin 5 expression in glands and ducts is clearly visible. TMEM16A immunoreactivity (goat anti-TMEM16A) was not present in Bowman’s glands of TMEM16A-/- nor of TMEM16A+/+ littermate mice (A, D). However, aquaporin 5 and TMEM16A were co-expressed in nasal septal glands (B) and lateral nasal glands (C) of TMEM16A+/+ mice. No immunoreactivity to TMEM16A was detectable in TMEM16A-/- mice (E, F). Glands marked by aquaporin 5 were similar in both types of mice. Images are averages of z-stacks of thickness of ~2.0 μm for A, D, or ~1 μm for B, C, E, F. Cell nuclei were stained by DAPI. Scale bars = 10 μm.
Mentions: Fig 8A and 8D shows that aquaporin 5 stained the internal wall of the duct of a Bowman’s gland as well as microvilli of supporting cells. Previous reports showed expression of TMEM16A in the duct of Bowman’s glands and in the luminal surface of nasal septal glands and lateral nasal glands [16]. However, in TMEM16A+/+ mice, we did not find expression of TMEM16A in Bowman’s glands, whereas the luminal surface of nasal septal glands and lateral nasal glands expressed both TMEM16A and aquaporin 5 (Fig 8B and 8C). A comparison with results from TMEM16A-/- mice shows that immunostaining of aquaporin 5 remains unchanged, whereas TMEM16A immunoreactivity was absent (Fig 8D–8F). The morphology of Bowman’s gland, nasal septal glands and lateral nasal glands remained the same in both types of mice.

Bottom Line: A comparison between the expression of the olfactory marker protein and adenylyl cyclase III shows that genetic ablation of TMEM16A did not seem to affect the maturation of olfactory sensory neurons and their ciliary layer.Moreover, we also observed that the morphology of Bowman's glands, nasal septal glands and lateral nasal glands did not change in the absence of TMEM16A.Our results indicate that the development of mouse olfactory epithelium and nasal glands does not seem to be affected by the genetic ablation of TMEM16A.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Olfactory Transduction, SISSA, International School for Advanced Studies, Trieste, Italy.

ABSTRACT
TMEM16A/ANO1 is a calcium-activated chloride channel expressed in several types of epithelia and involved in various physiological processes, including proliferation and development. During mouse embryonic development, the expression of TMEM16A in the olfactory epithelium is dynamic. TMEM16A is expressed at the apical surface of the entire olfactory epithelium at embryonic day E12.5 while from E16.5 its expression is restricted to a region near the transition zone with the respiratory epithelium. To investigate whether TMEM16A plays a role in the development of the mouse olfactory epithelium, we obtained the first immunohistochemistry study comparing the morphological properties of the olfactory epithelium and nasal glands in TMEM16A-/- and TMEM16A+/+ littermate mice. A comparison between the expression of the olfactory marker protein and adenylyl cyclase III shows that genetic ablation of TMEM16A did not seem to affect the maturation of olfactory sensory neurons and their ciliary layer. As TMEM16A is expressed at the apical part of supporting cells and in their microvilli, we used ezrin and cytokeratin 8 as markers of microvilli and cell body of supporting cells, respectively, and found that morphology and development of supporting cells were similar in TMEM16A-/- and TMEM16A+/+ littermate mice. The average number of supporting cells, olfactory sensory neurons, horizontal and globose basal cells were not significantly different in the two types of mice. Moreover, we also observed that the morphology of Bowman's glands, nasal septal glands and lateral nasal glands did not change in the absence of TMEM16A. Our results indicate that the development of mouse olfactory epithelium and nasal glands does not seem to be affected by the genetic ablation of TMEM16A.

No MeSH data available.


Related in: MedlinePlus