This research was supported by grants from the Research Foundation - Flanders (FWO, project Nr G.0420.02 and G.0443.11N), the Hercules Foundation (grant Nr AUHA0012), Concerted Research Actions (GOA funding) from the University of Antwerp, and partly sponsored by EC - FP6 project DiMI, LSHB-CT-2005-512146 and EC - FP6 project EMIL LSHC-CT-2004-503569 to A.VdL. G.DG and C.P. are Postdoctoral Fellows of the Research Foundation - Flanders (FWO).

1. Preparation of the Auditory Stimuli
<ol>
	<li>First record the sound-stimuli while being played inside the bore of the 7T MR system. The bore is a confined space that can distort the auditory stimuli resulting in enhancement of certain auditory frequencies. Figure 1 shows the frequencies enhanced and suppressed as shown by our recordings of white noise made at the location of the bird's head within the magnet bore using a fiber-optic microphone (Optimic 1160, Optoacoustics). To compensate this artif...

<ol>
	<li>Van Meir, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatiotemporal+properties+of+the+BOLD+response+in+the+songbirds'+auditory+circuit+during+a+variety+of+listening+tasks.">Spatiotemporal properties of the BOLD response in the songbirds' auditory circuit during a variety of listening tasks.</a> Neuroimage. 25, 1242-1255 (2005).</li><li>Boumans, T., Theunissen, F. E., Poirier, C., Van Der Linden, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+representation+of+spectral+and+temporal+features+of+song+in+the+auditory+forebrain+of+zebra+finches+as+revealed+by+functional+MRI.">Neural representation of spectral and temporal features of song in the auditory forebrain of zebra finches as revealed by functional MRI.</a> The European Journal of Neuroscience. 26, 2613-2626 (2007).</li><li>Boumans, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+magnetic+resonance+imaging+in+zebra+finch+discerns+the+neural+substrate+involved+in+segregation+of+conspecific+song+from+background+noise.">Functional magnetic resonance imaging in zebra finch discerns the neural substrate involved in segregation of conspecific song from background noise.</a> Journal of Neurophysiology. 99, 931-938 (2008).</li><li>Boumans, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+MRI+of+auditory+responses+in+the+zebra+finch+forebrain+reveals+a+hierarchical+organisation+based+on+signal+strength+but+not+selectivity.">Functional MRI of auditory responses in the zebra finch forebrain reveals a hierarchical organisation based on signal strength but not selectivity.</a> PloS ONE. 3, e3184 (2008).</li><li>Vignal, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+brain+hemodynamic+changes+in+a+songbird:+responses+to+hypercapnia+measured+with+functional+MRI+and+near-infrared+spectroscopy.">Measuring brain hemodynamic changes in a songbird: responses to hypercapnia measured with functional MRI and near-infrared spectroscopy.</a> Physics in Medicine and Biology. 53, 2457-2470 (2008).</li><li>Van der Linden, A., Van Meir, V., Boumans, T., Poirier, C., Balthazart, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI+in+small+brains+displaying+extensive+plasticity.">MRI in small brains displaying extensive plasticity.</a> Trends in Neurosciences. 32, 257-266 (2009).</li><li>Poirier, C., Van der Linden, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spin+echo+BOLD+fMRI+on+songbirds.">Spin echo BOLD fMRI on songbirds.</a> Methods Mol. Biol. 771, 569-576 (2011).</li><li>Poirier, C., Verhoye, M., Boumans, T., Van der Linden, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implementation+of+spin-echo+blood+oxygen+level-dependent+(BOLD)+functional+MRI+in+birds.">Implementation of spin-echo blood oxygen level-dependent (BOLD) functional MRI in birds.</a> NMR in Biomedicine. 23, 1027-1032 (2010).</li><li>Poirier, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+three-dimensional+MRI+atlas+of+the+zebra+finch+brain+in+stereotaxic+coordinates.">A three-dimensional MRI atlas of the zebra finch brain in stereotaxic coordinates.</a> Neuroimage. 41, 1-6 (2008).</li><li>Zhao, F., Wang, P., Kim, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+depth-dependent+gradient-echo+and+spin-echo+BOLD+fMRI+at+9.4T.">Cortical depth-dependent gradient-echo and spin-echo BOLD fMRI at 9.4T.</a> Magnetic Resonance in Medicine: Official Journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine. 51, 518-524 (2004).</li><li>Harel, N., Lin, J., Moeller, S., Ugurbil, K., Yacoub, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+imaging-histological+study+of+cortical+laminar+specificity+of+fMRI+signals.">Combined imaging-histological study of cortical laminar specificity of fMRI signals.</a> NeuroImage. 29, 879-887 (2006).</li><li>Duong, T. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microvascular+BOLD+contribution+at+4+and+7+T+in+the+human+brain:+gradient-echo+and+spin-echo+fMRI+with+suppression+of+blood+effects.">Microvascular BOLD contribution at 4 and 7 T in the human brain: gradient-echo and spin-echo fMRI with suppression of blood effects.</a> Magnetic Resonance in Medicine: Official Journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine. 49, 1019-1027 (2003).</li><li>Lee, S. P., Silva, A. C., Ugurbil, K., Kim, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diffusion-weighted+spin-echo+fMRI+at+9.4+T:+microvascular/tissue+contribution+to+BOLD+signal+changes.">Diffusion-weighted spin-echo fMRI at 9.4 T: microvascular/tissue contribution to BOLD signal changes.</a> Magnetic Resonance in Medicine: Official Journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine. 42, 919-928 (1999).</li><li>Uludag, K., Muller-Bierl, B., Ugurbil, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+integrative+model+for+neuronal+activity-induced+signal+changes+for+gradient+and+spin+echo+functional+imaging.">An integrative model for neuronal activity-induced signal changes for gradient and spin echo functional imaging.</a> NeuroImage. 48, 150-165 (2009).</li><li>Yacoub, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spin-echo+fMRI+in+humans+using+high+spatial+resolutions+and+high+magnetic+fields.">Spin-echo fMRI in humans using high spatial resolutions and high magnetic fields.</a> Magnetic Resonance in Medicine: Official Journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine. 49, 655-664 (2003).</li><li>Keilholz, S. D., Silva, A. C., Raman, M., Merkle, H., Koretsky, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+MRI+of+the+rodent+somatosensory+pathway+using+multislice+echo+planar+imaging.">Functional MRI of the rodent somatosensory pathway using multislice echo planar imaging.</a> Magnetic Resonance in Medicine: Official Journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine. 55, 316-324 (2006).</li><li>Keilholz, S. D., Silva, A. C., Raman, M., Merkle, H., Koretsky, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+MRI+of+the+rodent+somatosensory+pathway+using+multislice+echo+planar+imaging.">Functional MRI of the rodent somatosensory pathway using multislice echo planar imaging.</a> Magnetic Resonance in Medicine: Official Journal of the Society of Magnetic Resonance in Medicine / Society of Magnetic Resonance in Medicine. 52, 89-99 (2004).</li><li>Goloshevsky, A. G., Silva, A. C., Dodd, S. J., Koretsky, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=BOLD+fMRI+and+somatosensory+evoked+potentials+are+well+correlated+over+a+broad+range+of+frequency+content+of+somatosensory+stimulation+of+the+rat+forepaw.">BOLD fMRI and somatosensory evoked potentials are well correlated over a broad range of frequency content of somatosensory stimulation of the rat forepaw.</a> Brain Research. 1195, 67-76 (2008).</li><li>Kida, I., Yamamoto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulus+frequency+dependence+of+blood+oxygenation+level-dependent+functional+magnetic+resonance+imaging+signals+in+the+somatosensory+cortex+of+rats.">Stimulus frequency dependence of blood oxygenation level-dependent functional magnetic resonance imaging signals in the somatosensory cortex of rats.</a> Neuroscience Research. 62, 25-31 (2008).</li><li>Poirier, C., Boumans, T., Verhoye, M., Balthazart, J., Van der Linden, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Own-song+recognition+in+the+songbird+auditory+pathway:+selectivity+and+lateralization.">Own-song recognition in the songbird auditory pathway: selectivity and lateralization.</a> The Journal of Neuroscience: The Official Journal of the Society for Neuroscience. 29, 2252-2258 (2009).</li><li>Poirier, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Own+song+selectivity+in+the+songbird+auditory+pathway:+suppression+by+norepinephrine.">Own song selectivity in the songbird auditory pathway: suppression by norepinephrine.</a> PloS ONE. 6, e20131 (2011).</li><li>Logothetis, N. K., Pauls, J., Augath, M., Trinath, T., Oeltermann, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurophysiological+investigation+of+the+basis+of+the+fMRI+signal.">Neurophysiological investigation of the basis of the fMRI signal.</a> Nature. 412, 150-157 (2001).</li></ol>

No conflicts of interest declared.

In this report, we describe an optimized protocol for the detailed in vivo characterization of neural substrates of auditory stimulation in anaesthetized zebra finches.
In line with the presented protocol, the majority of functional brain activation studies in animals using BOLD fMRI, anaesthetize the animals during the acquisition. Training animals to accustom them to the magnet environment and the scanner noise during the study periods is also possible but rather time-consuming and ...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent/equipment</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments </td>
 </tr>
 <tr>
 <td>Isoflurane anaesthetic</td>
 <td>Isoflo</td>
 <td>05260-05</td>
 <td></td>
 </tr>
 <tr>
 <td>PC-Sam hardware/software</td>
 <td>SA-Instruments</td>
 <td></td>
 <td><a href="http://www.i4sa.com" target="_blank">http://www.i4sa.com</a></td>
 </tr>
 <tr>
 <td>Monitoring and gating system</td>
 <td></td>
 <td>1025</td>
 <td></td>
 </tr>
 <tr>
 <td>MR-compatible small rodent heater system</td>
 <td></td>
 <td></td>
 <td>Model 1025 compatible</td>
 </tr>
 <tr>
 <td>Rectal temperature probe</td>
 <td></td>
 <td>RTP-102B</td>
 <td>7'', 0.044'' </td>
 </tr>
 <tr>
 <td>7T MR scanner</td>
 <td>Bruker Biospin</td>
 <td>PHS 70/16 </td>
 <td></td>
 </tr>
 <tr>
 <td>Paravision software</td>
 <td></td>
 <td>5.1</td>
 <td></td>
 </tr>
 <tr>
 <td>Gradient Insert</td>
 <td></td>
 <td>BGA9S</td>
 <td>400 mT/m, 300A, 500V</td>
 </tr>
 <tr>
 <td>Gradient Amplifiers</td>
 <td>Copley Co., USA</td>
 <td>C256</td>
 <td></td>
 </tr>
 <tr>
 <td>Transmit resonators</td>
 <td></td>
 <td></td>
 <td>Inner diameter: 72 mm, transmit only, active decoupled</td>
 </tr>
 <tr>
 <td>Receiver antenna - 20 mm quadrature Mouse Head</td>
 <td></td>
 <td></td>
 <td>Receive only, active decoupled</td>
 </tr>
 <tr>
 <td>WaveLab software</td>
 <td>Steinberg</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Praat software</td>
 <td>Paul Boersma, University of Amsterdam</td>
 <td></td>
 <td><a href="http://www.praat.org" target="_blank">http://www.praat.org</a></td>
 </tr>
 <tr>
 <td>Non-magnetic dynamic speakers</td>
 <td>Visation, Germany</td>
 <td>HK 150</td>
 <td></td>
 </tr>
 <tr>
 <td>Fiber optic microphone</td>
 <td>Optoacoustics, </td>
 <td>Optimic 1160</td>
 <td></td>
 </tr>
 <tr>
 <td>Sound amplifier</td>
 <td>Phonic corporation</td>
 <td>MM 1002a</td>
 <td></td>
 </tr>
 <tr>
 <td>Presentation software</td>
 <td>Neurobehavioral Systems Inc.</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>MRIcro</td>
 <td>Chris Rorden</td>
 <td></td>
 <td><a href="http://www.cabiatl.com/mricro/mricro/" target="_blank">http://www.cabiatl.com/mricro/mricro/</a></td>
 </tr>
 <tr>
 <td>Statistical Parametric Mapping (SPM)</td>
 <td>Welcome Trust Centre for Neuroimaging</td>
 <td>8</td>
 <td><a href="http://www.fil.ion.ucl.ac.uk/spm/" target="_blank">http://www.fil.ion.ucl.ac.uk/spm/</a></td>
 </tr>
 </tbody>
</table>

functional magnetic resonance imaging (fmri) with auditory stimulation in songbirds

The neurobiology of birdsong, as a model for human speech, is a pronounced area of research in behavioral neuroscience. Whereas electrophysiology and molecular approaches allow the investigation of either different stimuli on few neurons, or one stimulus in large parts of the brain, blood oxygenation level dependent (BOLD) functional Magnetic Resonance Imaging (fMRI) allows combining both advantages, i.e. compare the neural activation induced by different stimuli in the entire brain at once. fMRI in songbirds is challenging because of the small size of their brains and because their bones and especially their skull comprise numerous air cavities, inducing important susceptibility artifacts. Gradient-echo (GE) BOLD fMRI has been successfully applied to songbirds 1-5 (for a review, see 6). These studies focused on the primary and secondary auditory brain areas, which are regions free of susceptibility artifacts. However, because processes of interest may occur beyond these regions, whole brain BOLD fMRI is required using an MRI sequence less susceptible to these artifacts. This can be achieved by using spin-echo (SE) BOLD fMRI 7,8 . In this article, we describe how to use this technique in zebra finches (Taeniopygia guttata), which are small songbirds with a bodyweight of 15-25 g extensively studied in behavioral neurosciences of birdsong. The main topic of fMRI studies on songbirds is song perception and song learning. The auditory nature of the stimuli combined with the weak BOLD sensitivity of SE (compared to GE) based fMRI sequences makes the implementation of this technique very challenging.

We here visually presented an optimized sequence of procedures for successful imaging of neural substrates of auditory stimuli in the zebra finch brain. Firstly, the described procedure for preparation of the auditory stimuli results in stimuli that can be incorporated into an ON/OFF block paradigm (Figure 2) and that are normalized to eliminate potential differences in sound pressure level that could evoke a differential response in the brain. After preparing the zebra finch for MRI scanning and ...

Watch this Scientific Journal Video about Functional Magnetic Resonance Imaging fMRI with Auditory Stimulation in Songbirds at JoVE.com

Functional Magnetic Resonance Imaging fMRI with Auditory Stimulation in Songbirds

This article shows an optimized procedure for imaging of the neural substrates of auditory stimulation in the songbird brain using functional Magnetic Resonance Imaging (fMRI). It describes the preparation of the sound stimuli, the positioning of the subject and the acquisition and subsequent analysis of the fMRI data.

Functional Magnetic Resonance Imaging (fMRI) with Auditory Stimulation in Songbirds

The neurobiology of birdsong, as a model for human speech, is a pronounced area of research in behavioral neuroscience. Whereas electrophysiology and ...

functional-magnetic-resonance-imaging-fmri-with-auditory-stimulation

Research

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Behavior

18.0K vistas.  University of Antwerp. En este artículo se muestra un procedimiento optimizado para obtener imágenes de los sustratos neurales de estimulación auditiva en el cerebro de un pájaro usando resonancia magnética funcional (fMRI). Se describe la preparación de los estímulos de sonido, el posicionamiento del sujeto y la adquisición y el análisis posterior de los datos de resonancia magnética funcional.