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Spontaneous and Acetylcholine Evoked Calcium Transients in the Developing Mouse Utricle.

Abstract

Spontaneous calcium transients are present during early postnatal development in the mouse retina and cochlea, and play an important role in maturation of the sensory organs and neural circuits in the central nervous system (CNS). It is not known whether similar calcium transients occur during postnatal development in the vestibular sensory organs. Here we demonstrate spontaneous intracellular calcium transients in sensory hair cells (HCs) and supporting cells (SCs) in the murine utricular macula during the first two postnatal weeks. Calcium transients were monitored using a genetically encoded calcium indicator, GCaMP5G (G5), at 100 ms-frame-1 in excised utricle sensory epithelia, including HCs, SCs, and neurons. The reporter line expressed G5 and tdTomato (tdT) in a Gad2-Cre dependent manner within a subset of utricular HCs, SCs and neurons. Kinetics of the G5 reporter limited temporal resolution to calcium events lasting longer than 200 ms. Spontaneous calcium transients lasting 1-2 s were observed in the expressing population of HCs at birth and slower spontaneous transients lasting 10-30 s appeared in SCs by P3. Beginning at P5, calcium transients could be modulated by application of the efferent neurotransmitter acetylcholine (ACh). In mature mice, calcium transients in the utricular macula occurred spontaneously, had a duration 1-2 s, and could be modulated by the exogenous application of acetylcholine (ACh) or muscarine. Long-lasting calcium transients evoked by ACh in mature mice were blocked by atropine, consistent with previous reports describing the role of muscarinic receptors expressed in calyx bearing afferents in efferent control of vestibular sensation. Large spontaneous and ACh evoked transients were reversibly blocked by the inositol trisphosphate receptor (IP3R) antagonist aminoethoxydiphenyl borate (2-APB). Results demonstrate long-lasting calcium transients are present in the utricular macula during the first postnatal week, and that responses to ACh mature over this same time period.

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  • Authors+Show Affiliations

    ,

    Department of Biomedical Engineering, University of Utah, Salt Lake City, UT, United States.

    ,

    Department of Biomedical Engineering, University of Utah, Salt Lake City, UT, United States. School of Biomedical Science and Pharmacy, Hunter Medical Research Institute, University of Newcastle, Newcastle, NSW, Australia.

    ,

    Department of Biomedical Engineering, University of Utah, Salt Lake City, UT, United States.

    Department of Biomedical Engineering, University of Utah, Salt Lake City, UT, United States. Neuroscience Program, University of Utah, Salt Lake City, UT, United States. Otolaryngology-Head and Neck Surgery, University of Utah, Salt Lake City, UT, United States.

    Source

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    31133810

    Citation

    Holman, Holly A., et al. "Spontaneous and Acetylcholine Evoked Calcium Transients in the Developing Mouse Utricle." Frontiers in Cellular Neuroscience, vol. 13, 2019, p. 186.
    Holman HA, Poppi LA, Frerck M, et al. Spontaneous and Acetylcholine Evoked Calcium Transients in the Developing Mouse Utricle. Front Cell Neurosci. 2019;13:186.
    Holman, H. A., Poppi, L. A., Frerck, M., & Rabbitt, R. D. (2019). Spontaneous and Acetylcholine Evoked Calcium Transients in the Developing Mouse Utricle. Frontiers in Cellular Neuroscience, 13, p. 186. doi:10.3389/fncel.2019.00186.
    Holman HA, et al. Spontaneous and Acetylcholine Evoked Calcium Transients in the Developing Mouse Utricle. Front Cell Neurosci. 2019;13:186. PubMed PMID: 31133810.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Spontaneous and Acetylcholine Evoked Calcium Transients in the Developing Mouse Utricle. AU - Holman,Holly A, AU - Poppi,Lauren A, AU - Frerck,Micah, AU - Rabbitt,Richard D, Y1 - 2019/05/07/ PY - 2018/12/03/received PY - 2019/04/12/accepted PY - 2019/5/29/entrez PY - 2019/5/28/pubmed PY - 2019/5/28/medline KW - GCaMP5G KW - calcium KW - hair cell KW - neuron KW - supporting cell KW - utricle SP - 186 EP - 186 JF - Frontiers in cellular neuroscience JO - Front Cell Neurosci VL - 13 N2 - Spontaneous calcium transients are present during early postnatal development in the mouse retina and cochlea, and play an important role in maturation of the sensory organs and neural circuits in the central nervous system (CNS). It is not known whether similar calcium transients occur during postnatal development in the vestibular sensory organs. Here we demonstrate spontaneous intracellular calcium transients in sensory hair cells (HCs) and supporting cells (SCs) in the murine utricular macula during the first two postnatal weeks. Calcium transients were monitored using a genetically encoded calcium indicator, GCaMP5G (G5), at 100 ms-frame-1 in excised utricle sensory epithelia, including HCs, SCs, and neurons. The reporter line expressed G5 and tdTomato (tdT) in a Gad2-Cre dependent manner within a subset of utricular HCs, SCs and neurons. Kinetics of the G5 reporter limited temporal resolution to calcium events lasting longer than 200 ms. Spontaneous calcium transients lasting 1-2 s were observed in the expressing population of HCs at birth and slower spontaneous transients lasting 10-30 s appeared in SCs by P3. Beginning at P5, calcium transients could be modulated by application of the efferent neurotransmitter acetylcholine (ACh). In mature mice, calcium transients in the utricular macula occurred spontaneously, had a duration 1-2 s, and could be modulated by the exogenous application of acetylcholine (ACh) or muscarine. Long-lasting calcium transients evoked by ACh in mature mice were blocked by atropine, consistent with previous reports describing the role of muscarinic receptors expressed in calyx bearing afferents in efferent control of vestibular sensation. Large spontaneous and ACh evoked transients were reversibly blocked by the inositol trisphosphate receptor (IP3R) antagonist aminoethoxydiphenyl borate (2-APB). Results demonstrate long-lasting calcium transients are present in the utricular macula during the first postnatal week, and that responses to ACh mature over this same time period. SN - 1662-5102 UR - https://www.unboundmedicine.com/medline/citation/31133810/Spontaneous_and_Acetylcholine_Evoked_Calcium_Transients_in_the_Developing_Mouse_Utricle L2 - https://dx.doi.org/10.3389/fncel.2019.00186 DB - PRIME DP - Unbound Medicine ER -