Integrative Paper 10/14
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FMI-43
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Small fluorescent lipophilic dye Passes through open-cell transduction channels and is trapped in the cytoplasm
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DEG/ENac
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Family of channels which may mediate touch sensation in nematodes
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Math1
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Transcription factor essential for hair-cell specification
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Hensen's cells
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Type of supporting cell adjacent to hair cells
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0203mt antibody
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antibody purified against entire C-terminus
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C-terminus
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the end of an amino acid chain (protein or polypeptide), terminated by a free carboxyl group (-COOH).
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0203fl antibody
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antibody purified against a small peptide representing conserved sequences within the C-terminus
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Conserved sequence
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Sequence of nucleic acids/amino acids that is found in a variety of otherwise unrelated places in the genome
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GFP::mmTRPA1
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fusion protein labelled by the TRPa1 antibody and an antibody to GFP driven by a CMV promoter
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Myosin 1c
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the adaptation motor - a component of the transduction apparatus
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Cadherin 23
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a tip-link protein - a component of the transduction apparatus
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Kinocilium
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a special type of cilium on the apex of hair cells located in the sensory epithelium of the vertebrate inner ear
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Pericuticular Zone
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Vesicle-rich region of the apical surface
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Stereocilia
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Resembling hair-like projections/are arranged in bundles of 30-300/hair-cell opens mechanically gated ion channels/are the mechanosensing (many mechanisms by which cells convert mechanical stimulus into electro-chemical activity) organelles of hair cells.
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La3+ ions
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Can remove tip links and abolish mechanosensitivty in hair cells (abolish transduction channel function) More effective than BAPTA, almost completely eliminated FM1-43 labeling in all hair cells
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Ca2+ chelator BAPTA
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Can remove tip links and abolish mechanosensitivity in hair cells (abolish transduction channel function) Less effective than La3+ ions, resisted by hair cells at edge of sensory epithelium
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Morpholino Oligodendrocytes (Morpholinos)
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Can be injected into zebrafish embryos to inhibit protein expression
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Danio rerio
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zebrafish
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trpa1
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partial sequence in zebrafish encoding homologue drTRPA1
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drTRPA1
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zebrafish homologue of TRPA1 apparently necessary for complete function or development of embryonic lateral line hair cells
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trpa2
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partial sequence in zebrafish encoding homologue drTRPA2
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drTRPA2
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zebrafish homologue of TRPA2??? probably not involved in hair-transduction
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h.p.f
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hours post-fertilization
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AM1-43
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fixable analogue of FM1-43 activity-dependent fluorescent nerve terminal probe
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morph1
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drTRPA1's translation initiation site
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morph3
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drTRPA1's splice junction causde a frameshift mutation and a stop codon insertion before drTRPA1's transmembrane domains by blocking splicing of intron 20-21 or by using a cryptic splicing donor site 22 base pairs to the normal donor site
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Streptomycin
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Channel blocker that abolishes otocyst microphonic potential
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Otocyst Microphonic Potential
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Extracellular voltage resulting from current flow through transduction channels Recorded to assay inner ear hair cells of fish
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Otocyst
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the developing inner ear
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Utricle
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A small, fluid-filled sac in the vestibular system above the saccule that responds to static positions of the head
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siRNAs
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Small-interfering RNAs Cleave mRNAs after differentiation
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siRNA-6 and siRNA-14
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Small-interfering RNAs targeting mmTRPA1 Under the control of the H1 promoter in the pSUPER vector, which were then inserted into adenoviruses
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Adenovirus
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DNA virus with conjunctivitis or diarrhea Readily infect hair cells
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Adenoviral Vector
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Unlike first-gen adenoviruses, does not damage hair cell transduction Multiply deleted of the E1, polymerase, and pTP genes
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Ad-GFP
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Control adenovirus encoding GFP alone (no siRNA)
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Ad-6 abd Ad-14-infected cells
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Had FM1-43 accumulation and whole-cell transduction currents significantly smaller than uninfected or Ad-GFP-infected controls
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Ruthenium red
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Blocks transduction channel of turtle and mouse hair cells, similar to block of TRPA1 channels expressed in bullfrog oocytes
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Heteromultimeric
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contain two or more different polypeptide chains, with each polypeptide encoded by a separate gene
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Ankyrin domains
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TRPA1 and TRPN1 (NOMPC) are unique among the TRP channel superfamily in possessing a large number of these in their amino termini Might bind to motor proteins that maintain tension on the channel. Alternatively, might bind to other signalling or modulatory proteins
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N terminus
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terminus at which synthesis begins
