Calretinin (CR) is a major calcium binding protein widely expressed in the CNS. However, its synaptic function remains largely elusive. At the auditory synapse of the endbulb of Held, CR is selectively expressed in different subtypes. Combining electrophysiology with immunohistochemistry, we investigated the synaptic transmission at the endbulb of Held synapses with and without endogenous CR expression in mature CBA/CAJ mice of either sex. Two synapse subtypes showed similar basal synaptic transmission, except a larger quantal size in CR-expressing synapses. During high-rate stimulus trains, CR-expressing synapses showed improved synaptic efficacy with significantly less depression and lower asynchronous release, suggesting more efficient exocytosis than non-CR-expressing synapses. Conversely, CR-expressing synapses had a smaller readily releasable pool size, which was countered by higher release probability and faster synaptic recovery to support sustained release during high-rate activity. EGTA-AM treatment did not change the synaptic transmission of CR-expressing synapses, but reduced synaptic depression and decreased asynchronous release at non-CR-expressing synapses, suggesting that CR helps to minimize calcium accumulation during high-rate activity. Both synapses express parvalbumin, another calcium-binding protein with slower kinetics and higher affinity than CR, but not calbindin. Furthermore, CR-expressing synapses only express the fast isoform of vesicular glutamate transporter 1 (VGluT1), while most non-CR-expressing synapses express both VGluT1 and the slower VGluT2, which may underlie their lagged synaptic recovery. The findings suggest that, paired with associated synaptic machinery, differential CR expression regulates synaptic efficacy among different subtypes of auditory nerve synapses to accomplish distinctive physiological functions in transmitting auditory information at high rates. CR is a major calcium-binding protein in the brain. It remains unclear how endogenous CR impacts synaptic transmission. We investigated the question at the large endbulb of Held synapses with selective CR expression and found that CR-expressing and non-CR-expressing synapses had similar release properties under basal synaptic transmission. During high-rate activity, however, CR-expressing synapses showed improved synaptic efficacy with less depression, lower asynchronous release, and faster recovery. Furthermore, CR-expressing synapses use exclusive VGluT1 to refill synaptic vesicles, while non-CR-expressing synapses use both VGluT1 and the slower isoform of VGluT2. Our findings suggest that CR may play significant roles in promoting synaptic efficacy during high-rate activity, and selective CR expression can differentially impact signal processing among different synapses.