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Titel: Quantitative Fluorescent in situ Hybridization Reveals Differential Transcription Profile Sharpening of Endocytic Proteins in Cochlear Hair Cells Upon Maturation
VerfasserIn: Huang, Guobin UdsID
Eckrich, Stephanie
Sprache: Englisch
In:
Titel: Frontiers in Cellular Neuroscience
Bandnummer: 15
Verlag/Plattform: Frontiers
Erscheinungsjahr: 2021
Freie Schlagwörter: fluorescent in situ hybridization
cochlea
hair cells
endocytosis
dynamin-1
dynamin-3
endophilinA1
terminal differentiation
DDC-Sachgruppe: 610 Medizin, Gesundheit
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: The organ of Corti (OC) comprises two types of sensory cells: outer hair cells (OHCs) and inner hair cells (IHCs). While both are mechanotransducers, OHCs serve as cochlear amplifiers, whereas IHCs transform sound into transmitter release. Reliable sound encoding is ensured by indefatigable exocytosis of synaptic vesicles associated with efficient replenishment of the vesicle pool. Vesicle reformation requires retrieval of vesicle membrane from the hair cell’s membrane via endocytosis. So far, the protein machinery for endocytosis in pre-mature and terminally differentiated hair cells has only partially been deciphered. Here, we studied three endocytic proteins, dynamin1, dynamin-3, and endophilin-A1, by assessing their transcription profiles in pre-mature and mature mouse OCs. State-of-the-art RNAscope R fluorescent in situ hybridization (FISH) of whole-mount OCs was used for quantification of target mRNAs on singlecell level. We found that pre-mature IHCs contained more mRNA transcripts of dnm1 (encoding dynamin-1) and sh3gl2 (endophilin-A1), but less of dnm3 (dynamin3) than OHCs. These differential transcription profiles between OHCs and IHCs were sharpened upon maturation. It is noteworthy that low but heterogeneous signal numbers were found between individual negative controls, which highlights the importance of corresponding analyses in RNAscope R assays. Complementary immunolabeling revealed strong expression of dynamin-1 in the soma of mature IHCs, which was much weaker in pre-mature IHCs. By contrast, dynamin-3 was predominantly found in the soma and at the border of the cuticular plates of pre-mature and mature OHCs. In summary, using quantitative RNAscope R FISH and immunohistochemistry on wholemount tissue of both pre-mature and mature OCs, we disclosed the cellular upregulation of endocytic proteins at the level of transcription/translation during terminal differentiation of the OC. Dynamin-1 and endophilin-A1 likely contribute to the strengthening of the endocytic machinery in IHCs after the onset of hearing, whereas expression of dynamin-3 at the cuticular plate of pre-mature and mature OHCs suggests its possible involvement in activity-independent apical endocytosis.
DOI der Erstveröffentlichung: 10.3389/fncel.2021.643517
URL der Erstveröffentlichung: https://doi.org/10.3389/fncel.2021.643517
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-425302
hdl:20.500.11880/38160
http://dx.doi.org/10.22028/D291-42530
ISSN: 1662-5102
Datum des Eintrags: 2-Aug-2024
Bezeichnung des in Beziehung stehenden Objekts: Supplementary Material
In Beziehung stehendes Objekt: https://www.frontiersin.org/api/v3/articles/643517/file/Data_Sheet_1.PDF/643517_supplementary-materials_datasheets_1_pdf/1?isPublishedV2=false
https://www.frontiersin.org/api/v3/articles/643517/file/Data_Sheet_2.PDF/643517_supplementary-materials_datasheets_2_pdf/1?isPublishedV2=false
https://www.frontiersin.org/api/v3/articles/643517/file/Data_Sheet_3.PDF/643517_supplementary-materials_datasheets_3_pdf/1?isPublishedV2=false
https://www.frontiersin.org/api/v3/articles/643517/file/Data_Sheet_4.PDF/643517_supplementary-materials_datasheets_4_pdf/1?isPublishedV2=false
https://www.frontiersin.org/api/v3/articles/643517/file/Data_Sheet_5.PDF/643517_supplementary-materials_datasheets_5_pdf/1?isPublishedV2=false
Fakultät: M - Medizinische Fakultät
Fachrichtung: M - Biophysik
Professur: M - Prof. Dr. Jutta Engel
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes



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