We thank Mr. Rodrigo de Souza for assistance in maintaining mouse colonies. L.A.V.M is a CAPES postdoctoral fellow. This work was supported by the grant PRONEX (FAPEMIG: APQ-00476-14).

Individually-housed male adult (8-12 weeks) C57BL/6 mice were used in this experiment. Animals received proper care before, during and after experimental procedures with food and water ad libitum. All procedures were approved by the animal ethics committee from Federal University of Minas Gerais (protocol number 59/2014).
1. Electrode Placement
<ol>
	<li>Sedating and fixating the animal onto the stereotaxic apparatus
	<ol>
		<li>Sterilize all the necessary surgical instruments. 
		Note. Surgical instruments were sterilized for 3 minutes at 440 &#176;C. Cotton swabs were autoclaved at 20 psi (pounds per square inch) at 121 &#176;C for 20 min.</li>
		<li>Adjust the thermal platform controller to 37 &#176;C.</li>
		<li>Weigh the animal and calculate the appropriate dose for anesthesia induction. Use a mixture of ketamine and xylazine at a dose of 100 mg/kg ketamine and 8 mg/kg xylazine, given intraperitoneally (needle size, 31 G). The animal should fall asleep within 2 to 3 min.</li>
		<li>Use an electric shaver or razor to shave down the surgical site.</li>
		<li>Place the animal onto the stereotaxic apparatus over the pre-warmed heating plate.</li>
		<li>Hold the animal&#39;s head and insert the tip ear bars into each of the animal&#39;s ears to fix it to the stereotaxic platform.</li>
		<li>Verify there is no lateral head shifting and little vertical movement after by slowly shifting the animal&#39;s head positioning.</li>
		<li>Gently slide the anesthesia mask over the mouse&#39;s nose and fix it in place by tightening the screw.</li>
		<li>Set the isoflurane to 1% with 1.0 L/min of O2.</li>
		<li>Apply eye ointment to the animal&#39;s eyes to prevent corneal drying during the surgery.</li>
	</ol>
	</li>
	<li>Attaching the implant to the animal&#39;s head
	<ol>
		<li>Use the cotton swabs to prepare the surgical site with three alternating scrubs of povidone-iodine (or 2% chlorhexidine) and 70% ethanol.</li>
		<li>Use a pair of tweezers to verify anesthesia depth by lightly squeezing the animal&#39;s toes and verifying the loss of animal&#39;s pedal withdrawal (toe pinch) reflex.</li>
		<li>Make an incision about 3 mm posterior to the animal&#39;s ear line and stop at the eye line. The incision site must have approximately 1 cm in length to be large enough to receive the implant.</li>
		<li>Gently scrape the cranium with a bone scraper to improve glue and cement adherence. Do this light handedly with the intention of creating micro scratches.</li>
		<li>Carefully position surgical hooks to the loose skin to maintain an open surgical field and free of obstructions such as skin and fur.</li>
		<li>Use a sterile cotton swab to dry the animal&#39;s scalp.</li>
		<li>Use a dissecting microscope to visualize the top of the animal&#39;s cranium.</li>
		<li>Attach a needle to the stereotaxic holder and locate the bregma. Position the needle directly above the animal&#39;s head slightly touching the bregma.</li>
		<li>Zero out all coordinates on the digital tracer and then raise the needle.</li>
		<li>Fix the tDCS implant on the stereotaxic holder. Position the implant over the animal&#39;s head and lower it slowly onto the region of interest using the proper stereotaxic coordinates.</li>
		<li>Use a needle to spread 1 drop (approximately 35 &#956;L) of super glue onto the implant&#39;s base.</li>
		<li>Slowly move the holder downwards until it touches the skull. Be sure that the implant base is entirely in contact with the surface.</li>
		<li>Prepare the surgical cement according to the manufacturer&#39;s instructions.</li>
		<li>After precise positioning, apply 3 thin, even layer of cement across the cranium and onto the lower portion of the implant. Apply drop per drop using an application brush. Layers must form a hill-shaped structure for further structural support of the implant.</li>
		<li>Leave the implant&#39;s screw thread clean of cement to allow a smooth, unobstructed connection.</li>
		<li>Allow each layer to dry for approximately 4 minutes.</li>
		<li>When dry, carefully remove the holder until it is completely detached from the implant. Always use extreme caution when handling the implant, since it may be accidentally extracted from the animal&#39;s skull.</li>
	</ol>
	</li>
	<li>Finishing surgery and post-surgical care
	<ol>
		<li>Hydrate the animal&#39;s skin in the incision site with a saline-soaked cotton swab.</li>
		<li>Coat the skin over the base of the tDCS implant.</li>
		<li>Use a pair of tweezers to bring the tissue together and close the incision with a drop of surgical tissue glue per 0.2 cm of tissue.</li>
		<li>Infiltrate 1-2% lidocaine in the incision site and underlying tissues.</li>
		<li>Hydrate the mouse with 500 &#181;L of lactate Ringer&#39;s solution subcutaneously.</li>
		<li>Place the mouse into a pre-warmed (37 &#176;C) clean, single-housed cage.</li>
		<li>Put a small dish with wet food pellets in the cage for easy access to food in the following hours.</li>
		<li>Register the animal&#39;s post-surgical weight.</li>
		<li>Give the animal ketoprofen (5 mg/kg) subcutaneously after the surgery and on the next 2 days.</li>
		<li>Monitor the recovery of the animal closely for at least 1 week. Assess any sign of distress, such as piloerection, lack of grooming, reduced locomotion, wound scratching and inflammation of the surgical site.</li>
	</ol>
	</li>
</ol>
2. tDCS Setup and Stimulation
<ol>
	<li>tDCS Setup (see Figure 2) 
	&#8203;Note. Make sure that the tDCS stimulator is fully charged.
	<ol>
		<li>Attach the anode and cathode cables to the tDCS stimulator and make them available near the stimulation site. Attach the pin-type electrode to the stereotaxic holder.</li>
		<li>Set the thermal platform to 37 &#176;C.</li>
		<li>Turn on the oxygen flowmeter on the inhalation anesthesia system to 1 L/min.</li>
		<li>Place the mouse into the anesthesia induction chamber.</li>
		<li>Turn on the isoflurane vaporizer to 3%. Allow the animal to undergo isoflurane effects for 4 min.</li>
		<li>While the animal is in the induction chamber, use a sterile syringe to fill the body electrode with 0.9% saline solution.</li>
		<li>Remove the animal from the induction chamber and position its chest over the body electrode.</li>
		<li>Gently slide the anesthesia mask over the mouse&#39;s nose and fix it in place. Lower the isoflurane output to 1.5%.</li>
		<li>Fill the implant and the pin-type electrode with saline and carefully attach them.</li>
		<li>Adjust stimulation time and current intensity.</li>
		<li>Verify the contact quality on the tDCS stimulator.Optimal contact goes from 7 to 10 on a 1 to 10 scale.</li>
	</ol>
	</li>
	<li>Stimulation
	<ol>
		<li>Start the stimulation.</li>
		<li>Observe the current ramping up for 30 s to the selected value and maintaining itself steady for the established time, then, at the end of the session ramping down again.</li>
		<li>Activate the sham button for control mice.</li>
		<li>Observe the current ramping up for 30 s to the selected value and then down to 1 for the rest of the stimulation period with a final ramp to the selected value at the end with a consecutive ramp down.</li>
		<li>Once the stimulation session is complete, carefully transfer the animal to a pre-warmed (37 &#176;C) cage for 10 min. 
		Note. Animals start to awaken after 3 min.</li>
		<li>Turn the inhalation anesthesia system off.</li>
	</ol>
	</li>
</ol>

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In recent years, neurostimulation techniques have been entering clinical practice as a promising procedure to treat neuropsychiatric disorders23. To reduce the constraint imposed by the lack of knowledge of the mechanisms of neurostimulation, we presented here a tDCS mouse model carrying an electrode that can target brain regions. Since the electrode is chronically implantable, this animal model enables the investigation of long-lasting biological effects evoked by tDCS (for at least 1 month) in c...

Transcranial Direct Current Stimulation (tDCS) is a non-invasive, low-cost, therapeutic technique, which focuses on neuronal modulation through the use of low-intensity continuous currents1. There are currently two setups (anodal and cathodal) for tDCS. While the anodal stimulation exerts a current electric field too weak to trigger action potentials, electrophysiology studies have shown that this method produces changes in synaptic plasticity2. For example, evidence shows that tDCS induces long-term potentiation (LTP) effects such as increased peak amplitude of the excitatory postsynaptic potentials3,4 and modulation of cortical excitability5.
Conversely, cathodal stimulation induces inhibition, resulting in membrane hyperpolarization6. A hypothesis for this mechanism is based on the physiological findings where tDCS is described to modulate action potential frequency and duration in the neuronal body3. Notably, this effect does not directly evoke action potentials, though it can shift the depolarization threshold and facilitate or hamper neuronal firing7. These contrasting effects have been previously demonstrated. For example, anodal and cathodal stimulation produced opposing effects in conditioned responses registered via electromyography activity in rabbits8. However, studies have also shown that prolonged anodal stimulation sessions may decrease excitability while increasing cathodal currents may lead to excitability, presenting self-contrasting effects3.
Both anodal and cathodal stimuli aggregate the use of electrode pairs. For example, in anodal stimulation, the &#34;active&#34; or &#34;anode&#34; electrode is placed over the brain region to be modulated whereas the &#34;reference&#34; or &#34;cathode&#34; electrode is situated over a region where the effect of current is assumed to be insignificant9. In the cathodal stimulation, electrode disposition is inverted. The stimulation intensity for effective tDCS depends on the current intensity and electrode dimensions, which affect the electric field differently10. In most published studies, the average current intensity is between 0.10 to 2.0 mA and 0.1 mA to 0.8 mA for human and mice, respectively6,11. Although the electrode size of 35 cm2 is typically used in humans, there is no proper understanding regarding electrode dimensions for rodents and a more thorough investigation is needed6.
tDCS has been proposed in clinical studies with the attempt of offering an alternative or complementary treatment for several neurological and neuropsychiatric disorders11 such as epilepsy12, bipolar disorder13, stroke5, major depression14, Alzheimer&#39;s disease15, multiple sclerosis16 and Parkinson&#39;s disease17. Despite growing interest in tDCS and its use in clinical trials, detailed cellular and molecular evoked alterations in brain tissue, short and long-lasting effects, as well as behavioral outcomes, are yet to be more deeply investigated18,19. Since a direct human approach to thoroughly study tDCS is not viable, the use of a tDCS animal model may offer valuable insights into the cellular and molecular events underlying the therapeutic mechanisms of tDCS due to the accessibility to the animal&#39;s brain tissue.
Available evidence is limited regarding tDCS models in mice. Most of the reported models used different implanting layouts, electrode dimensions, and materials. For example, Winkler et al. (2017) implanted the head electrode (Ag/AgCl, 4 mm in diameter) filled with saline and fixed it to the cranium with acrylic cement and screws20. Different from our approach, their chest electrode was implanted (platinum, 20 x 1.5 mm). Nasehi et al. (2017) used a procedure very similar to ours, although the thoracic electrode was made from a saline-soaked sponge (carbon filled, 9.5 cm2)21. Another study implanted both electrodes into the animal&#39;s head, which was achieved by using fixed plates and covering the animal&#39;s head with a hydrogel conductor22. Here, we describe a tDCS mouse model that uses a chronically implanted electrode through simple surgical procedures and tDCS setup (Figure 1).

<table border="0" cellpadding="0" cellspacing="0" style="width:1152px;" width="1152">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:457px;">BD Ultra-Fine 50U Syringe</td>
			<td style="width:175px;">BD</td>
			<td style="width:101px;">10033430026</td>
			<td style="width:419px;">For intraperitonially injection.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Shaver (Philips Multigroom)</td>
			<td>Philips (Brazil)</td>
			<td>QG3340/16</td>
			<td style="width:419px;">For surgical site trimming.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical Equipment</td>
			<td></td>
			<td></td>
			<td style="width:419px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Model 940 Small Animal Stereotaxic Instrument with Digital Display Console</td>
			<td>KOPF</td>
			<td>940</td>
			<td style="width:419px;">For animal surgical restriction and positioning.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Model 922 Non-Rupture 60 Degree Tip Ear Bars</td>
			<td>KOPF</td>
			<td>922</td>
			<td style="width:419px;">For animal surgical restriction and positioning.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cannula Holder</td>
			<td>KOPF</td>
			<td>1766-AP</td>
			<td style="width:419px;">For implant positioning.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Precision Stereo Zoom Binocular Microscope (III) on Boom Stand</td>
			<td>WPI</td>
			<td>PZMIII-BS</td>
			<td style="width:419px;">For bregma localization and implant positioning.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Temperature Control System Model&#160;</td>
			<td>KOPF</td>
			<td>TCAT-2LV</td>
			<td style="width:419px;">For animal thermal control.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cold Light Source&#160;</td>
			<td>WPI</td>
			<td>WA-12633</td>
			<td style="width:419px;">For focal brightness</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tabletop Laboratory Animal Anesthesia System with Scavenging</td>
			<td>VetEquip</td>
			<td>901820</td>
			<td style="width:419px;">For isoflurane delivery and safety.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">VaporGuard Activated Charcoal Adsorption Filter</td>
			<td>VetEquip</td>
			<td>931401</td>
			<td style="width:419px;">Delivery system safety measures.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Model 923-B Mouse Gas Anesthesia Head Holder</td>
			<td>KOPF</td>
			<td>923-B</td>
			<td style="width:419px;">For animal restriction and O2 and isoflurane delivery.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen regulator, E-cylinder&#160;</td>
			<td>VetEquip</td>
			<td>901305</td>
			<td style="width:419px;">For O2 regulation and delivery.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen hose &#8211; green&#160;</td>
			<td>VetEquip</td>
			<td>931503</td>
			<td style="width:419px;">For O2 and isoflurane delivery.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Infrared Sterilizer 800 &#186;C</td>
			<td>Marconi</td>
			<td>MA1201</td>
			<td style="width:419px;">For instrument sterilization.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical Instruments</td>
			<td></td>
			<td></td>
			<td style="width:419px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fine Scissors - ToughCut</td>
			<td>Fine Science Tools</td>
			<td>14058-11</td>
			<td style="width:419px;">For incision.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical Hooks</td>
			<td>INJEX</td>
			<td>1636</td>
			<td style="width:419px;">In House Fabricated - Used to clear the surgical site from skin and fur.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Standard Tweezers or Forceps</td>
			<td>-</td>
			<td>-</td>
			<td style="width:419px;">For skin grasping.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical Consumables</td>
			<td></td>
			<td></td>
			<td style="width:419px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vetbond</td>
			<td>3M</td>
			<td>SC-361931</td>
			<td style="width:419px;">For incision closing.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cement and Catalyzer KIT (Duralay)</td>
			<td>Reliance</td>
			<td>2OZ</td>
			<td style="width:419px;">For implant fixation.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile Cotton Swabs (Autoclaved)</td>
			<td>JnJ</td>
			<td>75U</td>
			<td style="width:419px;">For surgical site antisepsis.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">24 Well Plate (Tissue Culture Plate)</td>
			<td>SARSTEDT</td>
			<td>831,836</td>
			<td style="width:419px;">For cement preparation.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Application Brush</td>
			<td>parkell</td>
			<td>S286</td>
			<td style="width:419px;">For cement mixing and application.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pharmaceutics</td>
			<td></td>
			<td></td>
			<td style="width:419px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylazin (ANASEDAN 2%)</td>
			<td>Ceva Pharmaceutical (Brazil)</td>
			<td>P10160</td>
			<td style="width:419px;">For anesthesia induction.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketamine (DOPALEN 10%)</td>
			<td>Ceva Pharmaceutical (Brazil)</td>
			<td>P30101</td>
			<td style="width:419px;">For anesthesia induction.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane (100%)</td>
			<td>Crist&#225;lia (Brazil)</td>
			<td>100ML</td>
			<td style="width:419px;">For anesthesia maintenance.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lidocaine (XYLESTESIN 5%)</td>
			<td>Cristal Pharma</td>
			<td>-</td>
			<td style="width:419px;">For post-surgical care.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketoprofen (PROFENID 100 mg)</td>
			<td>Sanofi Aventis</td>
			<td>20ML</td>
			<td style="width:419px;">For post-surgical care.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ringer&#39;s Lactate Solution</td>
			<td>SANOBIOL LAB</td>
			<td>############</td>
			<td style="width:419px;">For post-surgical care.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TobraDex (Dexamethasone 1 mg/g)</td>
			<td>Alcon</td>
			<td>631</td>
			<td style="width:419px;">For eye lubrification and protection.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stimulation</td>
			<td></td>
			<td></td>
			<td style="width:419px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Animal Transcranial Stimulator</td>
			<td>Soterix Medical</td>
			<td>2100</td>
			<td style="width:419px;">For current generation.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pin-type electrode Holder (Cylindrical Holder Base)</td>
			<td>Soterix Medical</td>
			<td>2100</td>
			<td style="width:419px;">Electrode support (Implant).</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pin-type electrode (Ag/AgCl)</td>
			<td>Soterix Medical</td>
			<td>2100</td>
			<td style="width:419px;">For current delivery (electrode).&#160;</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;">Pin-type electrode cap</td>
			<td>Soterix Medical</td>
			<td>2100</td>
			<td style="width:419px;">For implant protection.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;">Body Electrode (Ag/AgCl Coated)</td>
			<td>Soterix Medical</td>
			<td>2100</td>
			<td style="width:419px;">For current delivery (electrode).&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Saline Solution (0.9%)</td>
			<td>FarmaX</td>
			<td>############</td>
			<td style="width:419px;">Conducting medium for current delivery.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Standard Tweezers or Forceps</td>
			<td>-</td>
			<td>-</td>
			<td style="width:419px;">For tDCS setup.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:457px;">Real Time Polymerase Chain Reaction</td>
			<td style="width:175px;"></td>
			<td style="width:101px;"></td>
			<td style="width:419px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:457px;">BioRad CFX96 Real Time System</td>
			<td style="width:175px;">BioRad</td>
			<td style="width:101px;">C1000</td>
			<td style="width:419px;">For qPCR</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SsoAdvancedTM Universal SYBR Green Supermix (5 X 1mL)</td>
			<td style="width:175px;">BioRad</td>
			<td>1725271</td>
			<td style="width:419px;">For qPCR</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hard Shell PCR Plates PCT COM 50 p/ CFX96</td>
			<td>BioRad</td>
			<td>HSP9601</td>
			<td style="width:419px;">For qPCR</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microseal &#34;B&#34; seal pct c/ 100</td>
			<td>BioRad</td>
			<td>MSB1001</td>
			<td style="width:419px;">For qPCR</td>
		</tr>
	</tbody>
</table>

transcranial direct current stimulation (tdcs) in mice

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for several neuropsychiatric diseases. The biological effects of tDCS are not fully understood, which is in part explained due to the difficulty in obtaining human brain tissue. This protocol describes a tDCS mouse model that uses a chronically implanted electrode allowing the study of the long-lasting biological effects of tDCS. In this experimental model, tDCS changes the cortical gene expression and offers a prominent contribution to the understanding of the rationale for its therapeutic use.

The surgical protocol presented long-term implant stability for at least one month, with no inflammatory signals at the stimulated site nor any other undesired effect. All the animals survived the surgical procedure and tDCS sessions (n = 8). In this experiment, tDCS implants were positioned over the M1 and M2 cortices (+1.0 mm anterior-posterior and 0.0 mm lateral to bregma). One week later, tDCS (n = 3-4) and sham (n = 3) mice were stimulated for five consecutive days during 10 min at 0...

Watch this Scientific Journal Video about Transcranial Direct Current Stimulation tDCS in Mice at JoVE.com

Transcranial Direct Current Stimulation tDCS in Mice

Transcranial direct current stimulation (tDCS) is a therapeutic technique proposed to treat psychiatric diseases. An animal model is essential for understanding the specific biological alterations evoked by tDCS. This protocol describes a tDCS mouse model that uses a chronically implanted electrode.

Transcranial Direct Current Stimulation (tDCS) in Mice

Transcranial direct current stimulation (tDCS) is a non-invasive neuromodulation technique proposed as an alternative or complementary treatment for ...

transcranial-direct-current-stimulation-tdcs-in-mice

Research

JoVE Journal

Neuroscience

13.9K Views.  Universidade Federal de Minas Gerais. Transcranial direct current stimulation (tDCS) is a therapeutic technique proposed to treat psychiatric diseases. An animal model is essential for understanding the specific biological alterations evoked by tDCS. This protocol describes a tDCS mouse model that uses a chronically implanted electrode.

Video: Transcranial Direct Current Stimulation tDCS in Mice

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful rest."1,2,3 It is thought the default mode network corresponds to self-referential or "internal mentation".2,3
It has been hypothesized that, in humans, activity within the default mode network is correlated with certain pathologies (for instance, hyper-activation has been linked to schizophrenia 4,5,6 and autism spectrum disorders 7 whilst hypo-activation of the network has been linked to Alzheimer's and other neurodegenerative diseases 8). As such, noninvasive modulation of this network may represent a potential therapeutic intervention for a number of neurological and psychiatric pathologies linked to abnormal network activation. One possible tool to effect this modulation is Transcranial Magnetic Stimulation: a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability (either increasing or decreasing it) via the application of localized magnetic field pulses.9
In order to explore the default mode network's propensity towards and tolerance of modulation, we will be combining TMS (to the left inferior parietal lobe) with functional magnetic resonance imaging (fMRI). Through this article, we will examine the protocol and considerations necessary to successfully combine these two neuroscientific tools.

In this article, we examine the methodology and considerations relevant to the combination of TMS and fMRI to examine the effects of brain stimulation on the default network.

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful ...

Electrode Positioning and Montage in Transcranial Direct Current Stimulation

Transcranial direct current stimulation (tDCS) is a technique that has been intensively investigated in the past decade as this method offers a non-invasive and safe alternative to change cortical excitability2. The effects of one session of tDCS can last for several minutes, and its effects depend on polarity of stimulation, such as that cathodal stimulation induces a decrease in cortical excitability, and anodal stimulation induces an increase in cortical excitability that may last beyond the duration of stimulation6. These effects have been explored in cognitive neuroscience and also clinically in a variety of neuropsychiatric disorders &#x2013; especially when applied over several consecutive sessions4. One area that has been attracting attention of neuroscientists and clinicians is the use of tDCS for modulation of pain-related neural networks3,5. Modulation of two main cortical areas in pain research has been explored: primary motor cortex and dorsolateral prefrontal cortex7. Due to the critical role of electrode montage, in this article, we show different alternatives for electrode placement for tDCS clinical trials on pain; discussing advantages and disadvantages of each method of stimulation.

Transcranial direct current stimulation (tDCS) is an established technique to modulate cortical excitability1,2. It has been used as an investigative tool in neuroscience due to its effects on cortical plasticity, easy operation, and safe profile. One area that tDCS has been showing encouraging results is pain alleviation 3-5.

Transcranial direct current stimulation (tDCS) is a technique that has been intensively investigated in the past decade as this method offers a ...

Direct-current Stimulation and Multi-electrode Array Recording of Seizure-like Activity in Mice Brain Slice Preparation

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the stimulation parameters (e.g., orientation, field strength, and stimulation duration) need to be examined in mice brain slice preparations. Testing and arranging the orientation of the electrode relative to the position of the mice brain slice are feasible. The present method preserves the thalamocingulate pathway to evaluate the effect of DCS on anterior cingulate cortex seizure-like activities. The results of the multichannel array recordings indicated that cathodal DCS significantly decreased the amplitude of the stimulation-evoked responses and duration of 4-aminopyridine and bicuculline-induced seizure-like activity. This study also found that cathodal DCS applications at 15 min caused long-term depression in the thalamocingulate pathway. The present study investigates the effects of DCS on thalamocingulate synaptic plasticity and acute seizure-like activities. The current procedure can test the optimal stimulation parameters including orientation, field strength, and stimulation duration in an in vitro mouse model. Also, the method can evaluate the effects of DCS on cortical seizure-like activities at both the cellular and network levels.

Studies have shown that cathodal transcranial direct-current stimulation can produce suppressive effects on drug-resistant seizures. In this study, an in vitro experimental setup was devised in which the direct-current stimulation and multielectrode array recording of seizure-like activity were evaluated in mice brain slice preparation. The direct-current stimulation parameters were evaluated.

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the ...

Effects of Transcranial Alternating Current Stimulation on the Primary Motor Cortex by Online Combined Approach with Transcranial Magnetic Stimulation

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific frequency and in turn modulate ongoing cortical oscillatory activity. This neurotool allows the establishment of a causal link between endogenous oscillatory activity and behavior. Most of the tACS studies have shown online effects of tACS. However, little is known about the underlying action mechanisms of this technique because of the AC-induced artifacts on Electroencephalography (EEG) signals. Here we show a unique approach to investigate online physiological frequency-specific effects of tACS of the primary motor cortex (M1) by using single pulse Transcranial Magnetic Stimulation (TMS) to probe cortical excitability changes. In our setup, the TMS coil is placed over the tACS electrode while Motor Evoked Potentials (MEPs) are collected to test the effects of the ongoing M1-tACS. So far, this approach has mainly been used to study the visual and motor systems. However, the current tACS-TMS setup can pave the way for future investigations of cognitive functions. Therefore, we provide a step-by-step manual and video guidelines for the procedure.

Transcranial Alternating Current Stimulation (tACS) allows the modulation of cortical excitability in a frequency-specific fashion. Here we show a unique approach which combines online tACS with single pulse Transcranial Magnetic Stimulation (TMS) in order to "probe" cortical excitability by means of Motor Evoked Potentials.

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific ...

Remotely Supervised Transcranial Direct Current Stimulation: An Update on Safety and Tolerability

The remotely supervised tDCS (RS-tDCS) protocol enables participation from home through guided and monitored self-administration of tDCS treatment while maintaining clinical standards. The current consensus regarding the efficacy of tDCS is that multiple treatment sessions are needed to observe targeted behavioral reductions in symptom burden. However, the requirement for patients to travel to clinic daily for stimulation sessions presents a major obstacle for potential participants, due to work or family obligations or limited ability to travel. This study presents a protocol that directly overcomes these obstacles by eliminating the need to travel to clinic for daily sessions.
This is an updated protocol for remotely supervised self-administration of tDCS for daily treatment sessions paired with a program of computer-based cognitive training for use in clinical trials. Participants only need to attend clinic twice, for a baseline and study-end visit. At baseline, participants are trained and provided with a study stimulation device, and a small laptop computer. Participants then complete the remainder of their stimulation sessions at home while they are monitored via videoconferencing software.
Participants complete computerized cognitive remediation during stimulation sessions, which may serve a therapeutic role or as a &#34;placeholder&#34; for other computer-based activity. Computers are enabled for real-time monitoring and remote control by study staff.
Outcome measures that assess feasibility and tolerance are administered remotely with the aid of visual analogue scales that are presented onscreen. Following completion of all RS-tDCS sessions, participants return to clinic for a study end visit in which all study equipment is returned.
Results support the safety, feasibility, and scalability of the RS-tDCS protocol for use in clinical trials. Across 46 patients, 748 RS-tDCS sessions have been completed. This protocol serves as a model for use in future clinical trials involving tDCS.

This manuscript provides an updated remote supervision protocol that enables participation in transcranial direct current stimulation (tDCS) clinical trials while receiving treatment sessions from home. The protocol has been successfully piloted in both patients with multiple sclerosis and Parkinson&#39;s disease.

The remotely supervised tDCS (RS-tDCS) protocol enables participation from home through guided and monitored self-administration of tDCS treatment while ...

Transcranial Electrical Brain Stimulation in Alert Rodents

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in humans is transcranial direct current stimulation (tDCS). Less frequently, transcranial alternating current stimulation (tACS) or transcranial random noise stimulation (tRNS), a specific form of tACS using an electrical current applied randomly within a pre-defined frequency range, is used. The increase of noninvasive electrical brain stimulation research in humans, both for experimental and clinical purposes, has yielded an increased need for basic, mechanistic, safety studies in animals. This article describes a model for transcranial electrical brain stimulation (tES) through the intact skull targeting the motor system in alert rodents. The protocol provides step-by-step instructions for the surgical set-up of a permanent epicranial electrode socket combined with an implanted counter electrode on the chest. By placing a stimulation electrode into the epicranial socket, different electrical stimulation types, comparable to tDCS, tACS, and tRNS in humans, can be delivered. Moreover, the practical steps for tES in alert rodents are introduced. The applied current density, stimulation duration, and stimulation type may be chosen depending on the experimental needs. The caveats, advantages, and disadvantages of this set-up are discussed, as well as safety and tolerability aspects.

This protocol describes a surgical set-up for a permanent epicranial electrode socket and an implanted chest electrode in rodents. By placing a second electrode into the socket, different types of transcranial electrical brain stimulation can be delivered to the motor system in alert animals through the intact skull.

Transcranial electrical brain stimulation can modulate cortical excitability and plasticity in humans and rodents. The most common form of stimulation in ...

Chronic Transcranial Electrical Stimulation and Intracortical Recording in Rats

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a closed-loop event-triggered manner. Although many studies are focusing on the possible benefits and side-effects of TES in healthy and pathologic brains, there are still many fundamental open questions regarding the mechanism of action of the stimulation. Therefore, there is a clear need for a robust and reproducible method to test the acute and the chronic effects of TES in rodents. TES can be combined with regular behavioral, electrophysiological, and imaging techniques to investigate neuronal networks in vivo. The implantation of transcranial stimulation electrodes does not impose extra constraints on the experimental design while it offers a versatile, flexible tool to manipulate brain activity. Here we provide a detailed, step-by-step protocol to fabricate and implant transcranial stimulation electrodes to influence brain activity in a temporally constrained manner for months.

This detailed protocol describes transcranial stimulation electrode placement on the temporal bone in order to investigate the short- and long-term effects of transcranial electrical stimulation in freely moving rats.

Transcranial electrical stimulation (TES) is a powerful and relatively simple approach to diffusely influence brain activity either randomly or in a ...

The Combined Use of Transcranial Direct Current Stimulation and Robotic Therapy for the Upper Limb

Neurologic disorders such as stroke and cerebral palsy are leading causes of long-term disability and can lead to severe incapacity and restriction of daily activities due to lower and upper limb impairments. Intensive physical and occupational therapy are still considered main treatments, but new adjunct therapies to standard rehabilitation that may optimize functional outcomes are being studied.
Transcranial direct current stimulation (tDCS) is a noninvasive brain stimulation technique that polarizes underlying brain regions through the application of weak direct currents through electrodes on the scalp, modulating cortical excitability. Increased interest in this technique can be attributed to its low cost, ease of use, and effects on human neural plasticity. Recent research has been performed to determine the clinical potential of tDCS in diverse conditions such as depression, Parkinson&#39;s disease, and motor rehabilitation after stroke. tDCS helps enhance brain plasticity and seems to be a promising technique in rehabilitation programs.
A number of robotic devices have been developed to assist in the rehabilitation of upper limb function after stroke. The rehabilitation of motor deficits is often a long process requiring multidisciplinary approaches for a patient to achieve maximum independence. These devices do not intend to replace manual rehabilitation therapy; instead, they were designed as an additional tool to rehabilitation programs, allowing immediate perception of results and tracking of improvements, thus helping patients to stay motivated.
Both tDSC and robot-assisted therapy are promising add-ons to stroke rehabilitation and target the modulation of brain plasticity, with several reports describing their use to be associated with conventional therapy and the improvement of therapeutic outcomes. However, more recently, some small clinical trials have been developed that describe the associated use of tDCS and robot-assisted therapy in stroke rehabilitation. In this article, we describe the combined methods used in our institute for improving motor performance after stroke.

The combined use of transcranial direct current stimulation and robotic therapy as an add-on for conventional rehabilitation therapy may result in improved therapeutic outcomes due to modulation of brain plasticity. In this article, we describe the combined methods used in our institute for improving motor performance after stroke.

Neurologic disorders such as stroke and cerebral palsy are leading causes of long-term disability and can lead to severe incapacity and restriction of ...

A Protocol for Transcranial Photobiomodulation Therapy in Mice

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide range of neurological and psychiatric disorders, and memory enhancement in age-related cognitive decline and neurodegenerative diseases. We describe a laboratory protocol for transcranial photobiomodulation therapy (PBMT) in mice. Aged BALB/c mice (18 months old) are treated with a 660 nm laser transcranially, once daily for 2 weeks. Laser transmittance data shows that approximately 1% of the incident red light on the scalp reaches a 1 mm depth from the cortical surface, penetrating the dorsal hippocampus. Treatment outcomes are assessed by two methods: a Barnes maze test, which is a hippocampus-dependent spatial learning and memory task evaluation, and measuring hippocampal ATP levels, which is used as a bioenergetics index. The results from the Barnes task show an enhancement of the spatial memory in laser-treated aged mice when compared with age-matched controls. Biochemical analysis after laser treatment indicates increased hippocampal ATP levels. We postulate that the enhancement of memory performance is potentially due to an improvement in hippocampal energy metabolism induced by the red laser treatment. The observations in mice could be extended to other animal models since this protocol could potentially be adapted to other species frequently used in translational neuroscience, such as rabbit, cat, dog, or monkey. Transcranial photobiomodulation is a safe and cost-effective modality which may be a promising therapeutic approach in age-related cognitive impairment.

Photobiomodulation therapy is an innovative noninvasive modality for the treatment of a wide range of neurological and psychiatric disorders and can also improve healthy brain function. This protocol includes a step-by-step guide to performing brain photobiomodulation in mice by transcranial light delivery, which can be adapted for use in other laboratory rodents.

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide ...

Stimulation Location Determination using a 3D Digitizer with High-Definition Transcranial Direct Current Stimulation

The abundance of neuroimaging data and rapid development of machine learning has made it possible to investigate brain activation patterns. However, causal evidence of brain area activation leading to a behavior is often left missing. Transcranial direct current stimulation (tDCS), which can temporarily alter brain cortical excitability and activity, is a noninvasive neurophysiological tool used to study causal relationships in the human brain. High-definition transcranial direct current stimulation (HD-tDCS) is a noninvasive brain stimulation (NIBS) technique that produces a more focal current compared to conventional tDCS. Traditionally, the stimulation location has been roughly determined through the 10-20 EEG system, because determining precise stimulation points can be difficult. This protocol uses a 3D digitizer with HD-tDCS to increase accuracy in determination of stimulation points. The method is demonstrated using a 3D digitizer for more accurate localization of stimulation points in the right temporo-parietal junction (rTPJ).

Presented here is a protocol to achieve higher accuracy in determination of stimulation location combining a 3D digitizer with high-definition transcranial direct current stimulation.

The abundance of neuroimaging data and rapid development of machine learning has made it possible to investigate brain activation patterns. However, ...