1Frogs must have sharp hearing abilities during the warm summer months to 2 successfully find mating partners. This study aims to understand how frog hair cell 3 ribbon-type synapses preserve both sensitivity and temporal precision during temperature 4 changes. We performed in vitro patch-clamp recordings of hair cells and their afferent 5 fibers in bullfrog amphibian papillae under room (23-25°C) and high (30-33°C) 6 temperature. Afferent fibers exhibited a wide heterogeneity in membrane input resistance 7 (R in ) from 100 MΩ to 1000 MΩ, which may contribute to variations in spike threshold 8 and firing frequency. At higher temperatures, most fibers increased their frequency of 9 action potential firing due to an increase in spontaneous EPSC frequencies. Hair cell 10 resting membrane potential (V rest ) remained surprisingly stable during temperature 11 increases, although both inward Ca 2+ current and outward K + current increased in 12 amplitude. This increase in Ca 2+ current may explain the higher spontaneous EPSC 13 frequencies. The larger "leak currents" at V rest lowered R in and produced higher electrical 14 resonant frequencies. However, lower R in should decrease sensitivity to sound detection 15 via smaller receptor potentials. Using membrane capacitance measurements, we suggest 16 that hair cells can partially compensate for this reduced sensitivity by increasing 17 exocytosis efficiency and the size of the readily releasable pool of synaptic vesicles. 18 Furthermore, paired recordings of hair cells and their afferent fibers showed that synaptic 19 delays become shorter and multivesicular release becomes more synchronous at higher 20 temperatures, which should improve temporal precision. Altogether, our results explain 21 many previous in vivo observations on the temperature dependence of spikes in auditory 22 nerves. 233
Significance Statement 24The vertebrate inner ear detects and transmits auditory information over a broad 25 dynamic range of sound frequency and intensity. It achieves remarkable sensitivity to soft 26 sounds and precise frequency selectivity. How does the ear of cold-blooded vertebrates 27 maintain its performance level as temperature changes? More specifically, how does the 28 hair cell to afferent fiber synapse in bullfrog amphibian papilla adjust to a wide range of 29 physiological temperatures without losing its sensitivity and temporal fidelity to sound 30 signals? This study uses in vitro experiments to reveal the biophysical mechanisms that 31 explain many observations made from in vivo auditory nerve fiber recordings. We find 32 that higher temperature facilitates vesicle exocytosis and electrical tuning to higher sound 33 frequencies, which benefits sensitivity and selectivity. 34Hair cells transduce sound vibrations into graded electrical signals, which are then 50 sent to the brain via all-or-none action potential spikes in the afferent fibers. At higher 51 temperatures, in vivo single afferent fiber recordings have revealed an increase in 52 spontaneous spike rates, a ...