Phase Relations of Purkinje Cells in the Rabbit Flocculus During Compensatory Eye Movements
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JOURNALOFNEUROPHYSIOLOGY Vol. 74, No. 5, November 1995. Printed in U.S.A. Phase Relations of Purkinje Cells in the Rabbit Flocculus During Compensatory Eye Movements C. I. DE ZEEUW, D. R. WYLIE, J. S. STAHL, AND J. I. SIMPSON Department of Physiology and Neuroscience, New York University Medical Center, New York 1OOM; and Department of Anatomy, Erasmus University Rotterdam, 3000 DR Rotterdam, Postbus 1738, The Netherlands SUMMARY AND CONCLUSIONS 17 cases (14%) showed CS modulation. In the majority (15 of 1. Purkinje cells in the rabbit flocculus that respond best to 17) of these cases, the CS activity increased with contralateral rotation about the vertical axis (VA) project to flocculus-receiving head rotation; these modulations occurred predominantly at the neurons (FRNs) in the medial vestibular nucleus. During sinusoi- higher stimulus velocities. dal rotation, the phase of FRNs leads that of medial vestibular 7. On the basis of the finding that FRNs of the medial vestibular nucleus neurons not receiving floccular inhibition (non-FRNs) . If nucleus lead non-FRNs, we predicted that floccular VA Purkinje the FRN phase lead is produced by signals from the ~~OCCU~US,then cells would in turn lead FRNs. This prediction is confirmed in the the Purkinje cells should functionally lead the FRNs. In the present present study. The data are therefore consistent with the hypothesis study we recorded from VA Purkinje cells in the flocculi of awake, that the phase-leading characteristics of FRN modulation could pigmented rabbits during compensatory eye movements to deter- come about by summation of VA Purkinje cell activity with that mine whether Purkinje cells have the appropriate firing rate phases of cells whose phase would otherwise be identical to that of non- to explain the phase-leading characteristics of the FRNs. FRNs. The floccular SS output appears to increase the phase lead 2. Awake rabbits were sinusoidally rotated about the VA in the of the net preoculomotor signal, which is in part composed of the light and the dark at 0.05-0.8 Hz with different amplitudes. The FRN and non-FRN signals. phase of the simple spike ( SS ) modulation in reference to eye 8. The phase lead of the floccular output may be created by and head position was calculated by determining the eye position emphasizing the velocity component of mossy fiber signals origi- sensitivity and the eye velocity sensitivity using multivariate linear nating in the vestibular nuclei. regression and Fourier analysis. The phase of the SS modulation in reference to head position was compared with the phase of the FRN modulation, which was obtained in prior experiments with INTRODUCTION the same stimulus paradigms. The cerebellar flocculus is involved in the control of eye 3. The SS activity of nearly all of the 88 recorded floccular VA movements. In rabbits, the main oculomotor behaviors con- Purkinje cells increased with contralateral head rotation. During rotation in the light, the SS modulation showed a phase lead in trolled by the flocculus are compensatory eye movements reference to contralateral head position that increased with increas- manifest as the vestibuloocular reflex (VOR) and the optoki- ing frequency (median 56.9” at 0.05 Hz, 78.6’ at 0.8 Hz). The SS netic reflex (OKR). The flocculus may contribute to the modulation led the FRN modulation significantly at all frequencies. control of both the gain and phase of compensatory eye The difference of medians was greatest (19.2”) at 0.05 Hz and movements. The possible short- and long-term roles of the progressively decreased with increasing frequency (all Ps < 0.005, flocculus in gain control have been extensively studied (rab- Wilcoxon rank-sum test). bit: Ito et al. 1982; Nagao 1983, 1989b; primate: Lisberger 4. During rotation in the dark, the SS modulation had a greater and Pavelko 1988; Lisberger et al. 1984; Takemori and Co- phase lead in reference to head position than in the light (median hen 1974; Waespe et al. 1983; Zee et al. 1981; chinchilla: 110.3’ at 0.05 Hz, 86.6’ at 0.8 Hz). The phase of the SS modulation Daniels et al. 1978; cat: Robinson 1976). Ito (Ito 1982; Ito in the dark led that of the FRNs significantly at all frequencies et al. 1974, 1982; see also Waespe and Henn 1981) initially (difference of medians varied from 24.2O at 0.05 Hz to 9.1” at 0.8 Hz; all Ps < 0.005). proposed that the flocculus augments OKR gain and medi- 5. The complex spike (CS) activity of all VA Purkinje cells ates enhancement of VOR gain by vision. Lesions of the increased with ipsilateral head rotation in the light. Fourier analy- flocculus and the ventral paraflocculus can change the gain sis of the cross-correlogram of the CS and SS activity showed of the VOR and OKR (Daniels et al. 1978; Ito et al. 1982; that the phase lag of the CS modulation in reference to the SS Lisberger et al. 1984; Nagao 1983, 1989b; Robinson 1976; modulation at 0.05 Hz in the light was not significantly different Takemori and Cohen 1974; Waespe et al. 1983; Zee et al. from that at 0.8 Hz (median 199.7’ at 0.05 Hz, 198.3O at 0.8 198 1) , and gain changes have been correlated with changes Hz), even though the phases of the SS modulation at these two in the simple spike (SS ) response of floccular Purkinje cells frequencies were significantly different (P < 0.001). These data (Nagao 1988, 1989b). indicate that the average temporal reciprocity between CS and SS modulation is fixed across the range of frequencies used in The role of the flocculus in the control of the phase of the present study. compensatory eye movements is less well documented. Eye 6. The CS activity of most Purkinje cells did not modulate movements in flocculectomized animals lag those of normals during rotation in the dark. Of 124 cases (each case consisting of during OKR and, to a lesser degree, during VOR (rabbit: the CS and SS data of a VA Purkinje cell obtained at 1 particular Ito et al. 1982; Nagao 1983; cat: Robinson 1976). Nagao frequency) examined over the frequency range of 0.05-0.8 Hz, ( 1983) reported that optokinetic stimulation in phase with 0022-3077/95 $3.00 Copyright 0 1995 The American Physiological Society 2051 2052 C. I. DE ZEEUW, D. R. WYLIE, J. S. STAHL, AND J. I. SIMPSON Light lead - 160 - FIG. 1. Summary of the phase relationships among different elements of the vestibuloocular re- flex (VOR) circuit (From Stahl and Simpson 1992, 1995b). The flocculus-receiving neurons (FRNs) lead the non-FRNs at all frequencies in both the JZIIl Nerve dark and the light. Phases (medians) are referenced to contralateral head position ( +, lead). For clarity of presentation, the phases of the medial vestibular FRN nucleus neurons (FRNs and non-FRNs) and primary afferents (VIIIth nerve) have been shifted by 180” nonFRN to reflect, respectively, their direct and indirect in- JZI hibitory action on the ipsilateral VIth nucleus neu- rons. VIth nucleus data were obtained in the light and the VIth nucleus phase values in the dark were obtained by referencing to eye position (Eye) in the dark. Eye 0.05 0.1 0.5 1.0 0.05 0.1 Stimulus Frequency (Hz) whole body rotation, but at twice the amplitude, induced a METHODS significant adaptive phase lead in the VOR that was abol- ished by bilateral lesion of floccular Purkinje cells with kai- Animal preparation nit acid. Injections of y-aminobutyric acid agonists into the Seven pigmented rabbits were prepared for chronic recording rabbit flocculus, on the other hand, influenced the gain but with the use of sterile surgical techniques. General anesthesia was not the phase of the VOR and OKR (Van Neerven et al. induced with a combination of ketamine (32 mg/kg im), acepro- 1989). The issue of a floccular contribution to phase control mazine (0.32 mg/ kg im) , and xylazine ( 5.0 mg/ kg om) , and sup- was also raised by Stahl and Simpson (1992, 1995b), who plemental doses (9 mg/kg ketamine, 0.09 mg/kg acepromazine, demonstrated in awake, pigmented rabbits that flocculus- 2 mg/kg xylazine) were given every 30-45 min. An acrylic head fixation pedestal was formed and fixed to the skull by screws receiving neurons ( FRNs ) in the medial vestibular nucleus implanted in the calvarium. The pedestal was oriented so that the have a phase lead with respect to head position that is sig- animal’s nasal bone made an angle of 57’ to the earth’s horizontal nificantly greater than that of other oculomotor projecting plane. With this orientation, the responses of the horizontal semicir- neurons in the medial vestibular nucleus that do not receive cular canals during rotation about the VA are close to their theoreti- an input from the flocculus (non-FRNs) . This phase differ- cal optimum, whereas the responses of the anterior and posterior ence was present throughout the tested frequency range canals are essentially zero (Soodak and Simpson 1988). A craniot- (0.05-0.8 Hz) for sinusoidal rotation about the vertical axis omy was made over the left paramedian lobule of the cerebellum, (VA) in both dark and light (Fig. 1) as well as for optoki- and a metal recording chamber to allow introduction of microelec- netic stimulation about the VA. This finding suggests that trodes into the flocculus was fixed around the craniotomy by ex- the phase lead of the FRNs over that of the non-FRNs could tending the acrylic head fixation pedestal. This cylindrical chamber be produced by the signal contributed by floccular Purkinje was oriented so that its axis was in a sagittal plane and made an angle of 27O to the vertical.