Immunogold EM quantification of calcium-buffering proteins in rat cochlear hair cells across development (P7, P16, P26). Primary source for the buffer_ratio parameter in h05 ODE models. The “Müller et al. 2002” attribution for buffer ratio 200-1000× in the h05 literature table is not found in the literature — the Hackney 2005 paper is the correct primary source. No Müller 2002 calcium buffer paper has been identified in any search.

Key finding

Mature rat OHC (P26) contains oncomodulin (parvalbumin-β) at 2-3 mM and calbindin-D28k at 15-230 µM (apex to base), totaling ~5-6 mM Ca2+-binding sites. OHC buffer capacity is ~110× that of IHC. Both proteins are partially excluded from stereocilia bundles, permitting faster unbuffered Ca2+ regulation at the MET channels. This supports a high cytoplasmic buffer ratio while the apical compartment (stereocilia) has lower buffering — relevant to h05 model parameter split between cytoplasm and stereocilia.

Numbers that matter

ParameterValueUnitsConditionsNotes
Oncomodulin (parvalbumin-β) in OHC1954–2892µMRat P26, apex to baseExpressed as immunogold-calibrated concentration
Calbindin-D28k in OHC15–230µMRat P26, basal to apicalLarge tonotopic gradient
Total Ca2+-binding sites in OHC~5–6mMP26, mature OHCEquivalent Ca2+-binding capacity
OHC/IHC buffer ratio~110×P26OHC has ~110× more proteinaceous Ca2+ buffer
Stereocilia vs cytoplasm3.5–5× lowerBoth parvalbumin-β and calbindinPartially excluded from stereocilia
Buffer ratio (κ, derived)~100–1000Ca2+-bound/Ca2+-freeModel-dependentHackney 2005 gives binding sites but not explicit κ; model value 1000 is upper end

Buffer ratio derivation: With 5 mM binding sites and Kd ~300 nM for calbindin, κ = [Ca-buffer]/[Ca-free] ≈ 5e-3 / (Kd + [Ca]×…). At [Ca] = 30 nM, most sites free; κ is high. The model range 100–1000× is consistent with Hackney 2005 if binding kinetics of oncomodulin (Kd ~1 µM) and calbindin (Kd ~300 nM) are applied.

Limitations

  • Immunogold EM gives relative, not absolute concentrations; calibration with known standards introduces uncertainty of ~20-30%.
  • Partial exclusion from stereocilia means OHC body values overestimate stereocilia buffer.
  • No “Müller 2002” paper found in any literature search — that attribution in h05 literature table is unverified phantom.

Connections