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. 2021 May 11;10:e63668. doi: 10.7554/eLife.63668

Figure 6. Differential development of cerebellar regions impacts the maturation of cerebellar-specific behaviors.

(A1) Schematic illustration of eye movement recording setup. Mice are head-fixed in the center of a turntable (green arrow) for vestibular stimulation and surrounded by a random dotted pattern drum (red arrow) for visual stimulation. A CCD camera was used for infrared (IR) video-tracking of the left eye (top left). Examples of nasal (N) and temporal (T) eye positions. Red circles = pupil fit; black cross = corneal reflection (CR); white cross = pupil (P) center. Example trace of eye position (gray) with drum position (red), during stimulation at an amplitude of 5° and frequency of 0.6 Hz (top right). Cerebellar circuitry controlling compensatory eye movements and their adaptation. Purkinje cells in the flocculus (FL) receive vestibular and visual input via the mossy fiber (MF) - parallel fiber (PF) system (green) and climbing fiber which influence eye movements via the vestibular nuclei (VN) and the oculomotor (OM) neurons. PN, pontine nuclei; GC, granule cell (bottom left). Photomicrograph of the flocculus a ZebrinII+ Purkinje cell predominant region; Purkinje cells labeled with Aldolase C (green) (bottom right). (A2) Results of 5 days of vestibule-ocular reflex (VOR) phase reversal training, probed by recording VOR (in the dark before, between and after sessions) with mice kept in the dark in between experimental sessions in young (P21-25, dotted line) and adult (P70-90, full line) mice. (B1) Schematic illustration of the eyeblink conditioning setup. Head-fixed mice on a freely moving treadmill, are presented a green LED light (conditioned stimulus, CS) followed several hundred milliseconds later by a weak air-puff on the eye (unconditioned stimulus, US). Eyelid movements were recorded with a camera (top). Cerebellar circuitry controlling eyeblink conditioning. Purkinje cells in the paravermal region around the primary fissure receive inputs carrying sensory information from for example the pontine nuclei (PN) through the mossy fiber-parallel fiber (MF-PF) pathway and the error signal from the inferior olive (IO) through the climbing fiber (CF). These Purkinje cells in turn influence eyelid muscles via the anterior interposed nucleus (AIN) and motor nuclei (MN) (bottom left). Photomicrograph of the base of paravermis a ZebrinII– Purkinje cell predominant region; Purkinje cells labeled with Aldolase C (green) (bottom right). (B2) As a result of repeated conditioned stimulus (CS)-unconditioned stimulus (US) pairings, mice will eventually learn to close their eye in response to the conditioned stimulus (CS), which is called the conditioned response (CR) (top). Percentage of conditioned response (CR%) in young (dotted line) and adult (full line) mice during 5 days of training (bottom). Error bars represent SEM., for values see Supplementary file 1. * denotes p<0.05, **p<0.001, and ***p<0.0001. Scale bars = (A) 200 µm.

Figure 6—source data 1. Eyeblink source data 1.
Figure 6—source data 2. Eye movement source data 2.
elife-63668-fig6-data2.xlsx (136.8KB, xlsx)

Figure 6.

Figure 6—figure supplement 1. Compensatory eye movements in young and adult mice.

Figure 6—figure supplement 1.

Gain (top) and phase (bottom) of baseline performance of compensatory eye movements: (A1 and B1) the optokinetic reflex (OKR), (A2 and B2) the vestibulo-ocular reflex (VOR) and (A3 and B3) the visually enhanced VOR (VVOR) in young (dotted line) and adult (full line) mice. Error bars represent SEM., for values see Supplementary file 1. * denotes p<0.05, **p<0.001 and ***p<0.0001.
Figure 6—figure supplement 2. Vestibulo-ocular reflex phase reversal gain, overnight consolidation, and gaze during training differ between juvenile and adult mice.

Figure 6—figure supplement 2.

(A) Example trace of eye movement position in young (blue) and adult (green) mice before and after training in relation with the table position (black) at an amplitude of 5°. (B) Gain of 5 days of vestibulo-ocular reflex (VOR) phase reversal, probed by recording VOR (in the dark before, between, and after sessions) with mice kept in the dark in between experimental sessions, in young (dotted line blue) and adult (full line green) mice. (C) Illustration of VOR gain consolidation analysis and the percentage of consolidation between young (blue) and adult (green) mice at days 1-2, 2–3, and 3–4. (D) Gain and (E) phase of 5 days of VOR phase reversal training, in young (dotted line blue) and adult (full line green) mice. Error bars represent SEM., for values see Supplementary file 1. * denotes p<0.05, **p<0.001, and ***p<0.0001.
Figure 6—figure supplement 3. Polar plot for vestibulo-ocular reflex phase reversal gain.

Figure 6—figure supplement 3.

Vestibulo-ocular reflex (VOR) phase reversal gain in (A) individual sessions and (B) averaged sessions per 5 days in young (blue) and adult (green) mice.
Figure 6—figure supplement 4. Eyeblink conditioning in young and adult mice.

Figure 6—figure supplement 4.

(A1) Example traces of a single unconditioned response and (A2) a single conditioned response. Insets: mouse eye video captures show eyelid closure ranging from 0 (fully-open) to 1 (fully-closed). (B1) The onset and (B2) peak time of the unconditioned response (UR) in young (dotted square) and adult (full square) mice. Comparison of fraction of eyelid closure between young (top) and adult mice (bottom). (C1 and C3) Waterfall plot showing the averaged eyeblink trace during conditioned stimulus (CS)-only trials for the 10 sessions (color intensity increasing from sessions 1 to 10). (C2 and C4) Session averages (thin gray lines) per mouse and overall average (thick black lines) for the 10 sessions.