BACKGROUND: Electrophysiological studies using brain slices have revealed that the developmental regulation of synaptic plasticity in vocal learning pathway is essential for song learning in zebra finches. Publications reporting in vivo electrophysiological investigation are scarce. Many aspects of neural mechanisms underlying song learning and production still remain uncertain. OBJECTIVE: To observe the efficacy of paired pulses and the effect on synaptic transmission induced by low-frequency stimulations, high-frequency stimulations, and theta-burst stimulations. DESIGN, TIME AND SETTING: A comparative observation. The experiment was conducted from October 2006 to October 2007 in the Neurophysiology Laboratory of South-China Normal University. MATERIALS: Twenty-four adult male zebra finches were supplied by the Department of Animal Experiment of College of Life Sciences, South China Normal University. A SEN-7203 stimulator (NIHON KOHDEN), as well as a DSJ-731WF microelectrode amplifier and DSJ-F amplifier (provided by South-China Normal University), were used to stimulate and record, respectively. METHODS: Animals were randomly divided into low-frequency, high-frequency, and theta-burst frequency stimulation groups. After recording evoked potentials, an input-output curve was evaluated. Subsequently, the efficacy of paired pulses with different stimulus intensity (1/3, 1/2, 2/3, or 3/4 of the value that induced the largest synaptic response), as well as interpulse intervals (50, 75, and 100 ms), was measured in each group. The test stimulus intensity was set to a level that evoked 1/2 or 1/3 amplitude of the maximum response. MAIN OUTCOME MEASURES: Changes in amplitude, slope, and area of evoked potentials elicited by different stimulus patterns. RESULTS: (1) Efficacy of paired pulses: there was significant paired-pulse facilitation in the high vocal center and robust nucleus of the arcopallium (HVC-RA) synapse. Efficacy decreased when paired-pulse intervals or stimulus intensities were increased. (2) Low-frequency stimulations at 1 Hz had a negligible effect on efficacy of synaptic transmission, while 5 Hz depressed synaptic transmission for only 5 minutes. (3) High-frequency stimulations at 50 Hz or 100 Hz induced synaptic depression that lasted for up to 30 minutes. (4) Theta-burst stimulation depressed synaptic transmission efficiency significantly for about 10 minutes. However, in contrast to low-frequency or high-frequency stimulations, theta-burst stimulations also induced slight potentiation of synaptic transmission for up to 60 minutes following depression phase, although the slope or area did not change. CONCLUSION: Paired pulses induced a remarkable efficacy of paired pulses in the high vocal center and robust nucleus of the arcopallium pathway. Low-frequency, high-frequency, or theta-burst frequency stimulation did not induce long-lasting changes in synaptic transmission.