Impaired reach-to-grasp responses in mice depleted of striatal cholinergic interneurons

by Nilupaer Abudukeyoumu, Marianela Garcia-Munoz, Yoko Nakano, Gordon W. Arbuthnott.
Brain Mechanism for Behavior Unit, Okinawa Institute of Science and Technology Graduate University, Okinawa, Japan

Cholinergic interneurons (ChIs) are sparsely distributed within the striatum, a nucleus that plays important roles in voluntary motor control, associated learning, procedural memory, execution of movement, action selection, and planning.[1]

ChIs comprise 1-3% of all striatal neurons, are the main source of striatal acetylcholine, and have long been associated with deficits in Parkinson’s disease. A striatal imbalance between dopamine and acetylcholine has been suggested as one of the causes of parkinsonism.[2]

To selectively deplete ChIs in the dorsolateral striatum of 21-day-old male mice (C57BL/6J), we used the saporin immunotoxin that targets choline acetyltransferase (Anti-ChAT-SAP, Cat. #IT-42). Experimental animals received a stereotaxic unilateral infusion of the targeted toxin (0.3μl/3min) and sham controls received the same volume of sterile saline (sham control). Our histological analysis encompassed Weeks 2-6 postsurgery performed at 2-week intervals (Fig. 1). The loss of cells reached a stable ~70% level by 4 to 6 weeks with the additional surprising finding that axon terminals stained with a vesicular acetylcholine transporter antibody were more numerous two weeks after the injection, returning to control levels by six weeks.

Figure 1.

From a functional point of view, it will be important to find out if despite the cell loss, axon terminals sprout to invade the Anti-ChAT-SAP injected area from ~30% surviving ChIs or from ChIs in the surrounding tissue. To begin the study of dorsolateral striatal function following Anti-ChAT-SAP-induced ChI loss, we followed the same procedures as before[3] and observed the animal’s perfomance in a reach-to-grasp task (Fig. 2). Mice were divided in two control groups (intact and sham) and one experimental Anti-ChAT-SAP-injected group. Training started one week postsurgery during the animal’s active circadian cycle and following 12 hours of food deprivation.

Figure 2. Damage to ChIs impaired the use of t

Once the animals passed the initial acquisition phase, the successful performance in the reach-to-grasp task — expressed as mean ± SD percentage is shown in Fig. 3.
Controls: 51.11 ± 3.83; n = 25 [intact], 48.79 ± 4.6; n = 9 [sham]
Treated: 26.28 ± 3.74; n = 13

The significantly-impaired performance of the experimental group compared to controls was present even when the animals were pretrained. The loss of ChIs impairs the performance of striatally-mediated motor tasks, which suggests that cholinergic synaptic function is more important than non-synaptic communication in this situation. A non-synaptic cholinergic tone may be important for setting functional striatal states in other circumstances,[4] however, these specific lesions of ChI cells suggest that performance of a learned forelimb task requires that the cholinergic synaptic circuits of the striatum are intact.


  1. Abudukeyoumu N, Hernández-Flores T, Garcia M, Arbuthnott, G. Cholinergic modulation of striatal microcircuits. (2018). Eur J Neurosci. 10.1111/ejn.13949.
  2. Aosaki T, Miura M, Suzuki T, Nishimura K, & Masuda M. Acetylcholine-dopamine balance hypothesis in the striatum: an update. (2010). Geriatr Gerontol Int., 10 Suppl 1 S148-157. 2010/07/16.
  3. Lopez-Huerta VG, Nakano Y, Bausenwein J, Jaidar O, Lazarus M, Cherassse Y, Garcia-Munoz M, & Arbuthnott G. The neostriatum: two entities, one structure? (2016). Brain Struct Funct, 221 (3):1737-1749. 2015/02/06. PMC4819794.
  4. Pittman-Polletta BR, Quach A, Mohammed AI, Romano M, Kondabolu K, Kopell NJ, Han X, & McCarthy MM. Striatal cholinergic receptor activation causes a rapid, selective and state-dependent rise in cortico-striatal beta activity. (2018). Eur J Neurosci. 48 (8):2857-2868.

Other References Using Anti-ChAT-SAP
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Abudukeyoumu N, Garcia-Munoz M, Jaidar OP, Arbuthnott G (2016) Striatal cholinergic interneurons: their depletion and its progression. Soc Neurosci Meeting Abstract 245.09
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