Flavoprotein fluorescence imaging-based electrode implantation for subfield-targeted chronic recording in the mouse auditory cortex.

BACKGROUND: Chronic neural recording in freely moving animals is important for understanding neural activities of cortical neurons associated with various behavioral contexts. In small animals such as mice, it has been difficult to implant recording electrodes into exact locations according to stereotactic coordinates, skull geometry, or the shape of blood vessels. The main reason for this difficulty is large individual differences in the exact location of the targeted brain area. NEW METHODS: We propose a new electrode implantation procedure that is combined with transcranial flavoprotein fluorescence imaging. We demonstrate the effectiveness of this method in the auditory cortex (AC) of mice. RESULTS: Prior to electrode implantation, we executed transcranial flavoprotein fluorescence imaging in anesthetized mice and identified the exact location of AC subfields through the skull in each animal. Next, we surgically implanted a microdrive with a tungsten electrode into exactly the identified location. Finally, we recorded neural activity in freely moving conditions and evaluated the success rate of recording auditory responses. COMPARISON WITH EXISTING METHOD(S): These procedures dramatically improved the success rate of recording auditory responses from 21.1% without imaging to 100.0% with imaging. We also identified large individual differences in positional relationships between sound-driven response areas and the squamosal suture or blood vessels. CONCLUSIONS: Combining chronic electrophysiology with transcranial flavoprotein fluorescence imaging before implantation enables the realization of reliable subfield-targeted neural recording from freely moving small animals.

Salicylate-induced frequency-map reorganization in four subfields of the mouse auditory cortex.

Salicylate is the active ingredient in aspirin, and in high-doses it is used as an experimental tool to induce transient hearing loss, tinnitus, and hyperacusis. These salicylate-induced perceptual disturbances are associated with tonotopic-map reorganization and neural activity modulation, and such neural correlates have been examined in the central auditory pathway, including the auditory cortex (AC). Although previous studies have reported that salicylate induces increases in noise-burst-evoked neural responses and reorganization of tonotopic maps in the primary AC, little is known about the effects of salicylate on other frequency-organized AC subfields such as the anterior auditory, secondary auditory, and dorsomedial fields. Therefore, to examine salicylate-induced spatiotemporal effects on AC subfields, we measured sound-evoked neural activity in mice before and after the administration of sodium salicylate (SS, 200 mg/kg), using flavoprotein auto-fluorescence imaging. SS-treatment gradually reduced responses driven by tone-bursts with lower (≤8 kHz) and higher (≥25 kHz) frequencies over 3 h, whereas evoked responses to tone-bursts within middle-range frequencies (e.g., 12 and 16 kHz) were sustained and unchanged in the four subfields. Additionally, in each of the four subfields, SS-treatment induced similar reorganization of tonotopic maps, and the response areas selectively driven by the middle-range frequencies were profoundly expanded. Our results indicate that the SS-induced tonotopic map reorganizations in each of the four AC subfields were similar, and only the extent of the activated areas responsive to tone-bursts with specific frequencies was subfield-dependent. Thus, we expect that examining cortical reorganization induced by SS may open the possibility of new treatments aimed at altering cortical reorganization into the normative functional organization.

Transcranial flavoprotein-autofluorescence imaging of sound-evoked responses in the mouse auditory cortex under three types of anesthesia.

The effects of anesthesia on the functional auditory characteristics of cortical neurons, such as spatial and temporal response properties, vary between an anesthetized and an awake subject. However, studies have shown that an appropriate anesthetic method that approaches the awake condition is still useful because of its greater stability and controllability. The present study compared neural response properties from two core fields of the mouse auditory cortex under three anesthetic conditions: urethane; ketamine and xylazine hydrochloride (KX) mixture; and a combination of medetomidine, midazolam, and butorphanol (MMB). To measure sound stimulation in vivo, we recorded flavoprotein-autofluorescent images of endogenous green fluorescence. Under all conditions, fluorescence changes in auditory core subfields in response to tones were observed, and response properties, such as peak intensity, latency, duration, and activated areas were analyzed. Results showed larger response peak intensity, latency, and duration in the core subfields under urethane compared with KX and MMB, with no significant differences between KX and MMB. Conversely, under KX anesthesia the activated areas showed characteristic response properties in a subfield-dependent manner. These results demonstrated the varied effects of anesthesia on response properties in the core subfields of the auditory cortex.