title: Chronic assessment of cerebral hemodynamics during rat forepaw electrical stimulation using functional ultrasound imaging.
abstract: Functional ultrasound imaging is a method recently developed to assess brain activity via hemodynamics in rodents. Doppler ultrasound signals allow the measurement of cerebral blood volume (CBV) and red blood cells' (RBCs') velocity in small vessels. However, this technique originally requires performing a large craniotomy that limits its use to acute experiments only. Moreover, a detailed description of the hemodynamic changes that underlie functional ultrasound imaging has not been described but is essential for a better interpretation of neuroimaging data. To overcome the limitation of the craniotomy, we developed a dedicated thinned skull surgery for chronic imaging. This procedure did not induce brain inflammation nor neuronal death as confirmed by immunostaining. We successfully acquired both high-resolution images of the microvasculature and functional movies of the brain hemodynamics on the same animal at 0, 2, and 7 days without loss of quality. Then, we investigated the spatiotemporal evolution of the CBV hemodynamic response function (HRF) in response to sensory-evoked electrical stimulus (1 mA) ranging from 1 (200 μs) to 25 pulses (5s). Our results indicate that CBV HRF parameters such as the peak amplitude, the time to peak, the full width at half-maximum and the spatial extent of the activated area increase with stimulus duration. Functional ultrasound imaging was sensitive enough to detect hemodynamic responses evoked by only a single pulse stimulus. We also observed that the RBC velocity during activation could be separated in two distinct speed ranges with the fastest velocities located in the upper part of the cortex and slower velocities in deeper layers. For the first time, functional ultrasound imaging demonstrates its potential to image brain activity chronically in small animals and offers new insights into the spatiotemporal evolution of cerebral hemodynamics.
authors.affiliation: Université Paris Descartes Sorbonne Paris Cité, Centre de Psychiatrie et Neurosciences, INSERM S894, Centre Hospitalier Sainte-Anne, Paris, France. Electronic address: alan.urban@alan-urban.fr.
authors.forename: Alan
authors.initials: A
authors.lastname: Urban
authors.affiliation: 1A Allée des bois de Gagny, 93340 Le Raincy, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Mace
authors.affiliation: Université Paris Descartes Sorbonne Paris Cité, Centre de Psychiatrie et Neurosciences, INSERM S894, Centre Hospitalier Sainte-Anne, Paris, France.
authors.forename: Clément
authors.initials: C
authors.lastname: Brunner
authors.affiliation: Université Paris Descartes Sorbonne Paris Cité, Centre de Psychiatrie et Neurosciences, INSERM S894, Centre Hospitalier Sainte-Anne, Paris, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Heidmann
authors.affiliation: Université Paris Descartes Sorbonne Paris Cité, Centre de Psychiatrie et Neurosciences, INSERM S894, Centre Hospitalier Sainte-Anne, Paris, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Rossier
authors.affiliation: Université Paris Descartes Sorbonne Paris Cité, Centre de Psychiatrie et Neurosciences, INSERM S894, Centre Hospitalier Sainte-Anne, Paris, France.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Montaldo
mesh_terms: D000818:Animals; D001810:Blood Volume / Q000502:physiology; D001921:Brain / Q000098:blood supply / Q000502:physiology*; D002540:Cerebral Cortex / Q000098:blood supply / Q000000981:diagnostic imaging / Q000502:physiology; D004558:Electric Stimulation / Q000379:methods; D004912:Erythrocytes / Q000000981:diagnostic imaging; D005073:Evoked Potentials, Somatosensory / Q000502:physiology*; D005552:Forelimb / Q000502:physiology; D059907:Functional Neuroimaging / Q000379:methods*; D006439:Hemodynamics / Q000502:physiology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012886:Skull / Q000601:surgery; D017585:Ultrasonography, Doppler, Transcranial / Q000379:methods*
keywords: Cerebral blood volume (CBV); Doppler ultrasound; Functional imaging; Neurovascular coupling; Thinned skull