Taiko Kitamura

Differences of the primate flocculus and ventral paraflocculus in the mossy and climbing fiber input organization

Potential sources of cerebellar cortical afferent fibers were identified in the vestibular ganglion, medulla oblongata, pons, and cerebellar nucleus of seven anesthetized Macaca fuscata after local injections of wheat germ agglutinin‐conjugated horseradish peroxidase or Fast Blue into the flocculus (FL) or ventral paraflocculus (VP). There were differences in the sources of mossy fibers to the FL and VP. Labeled neurons, after injections into the FL, were located mainly in the ipsilateral vestibular ganglion, bilaterally in the vestibular and prepositus hypoglossal nuclei, nucleus reticularis tegmenti pontis, and the central part of the mesencephalic reticular formation including the raphe nuclei. Labeled neurons were rarely seen in the pontine nuclei after injections into the FL. By contrast, after injections into the VP, numerous labeled neurons were located in the contralateral pontine nuclei, but relatively few in the vestibular nuclei bilaterally. Sources of climbing fibers to the FL and VP were completely contralateral to the injection side. After the injection into the FL and VP, labeled neurons were located in the dorsal cap, ventrolateral outgrowth, and ventral part of the medial accessory olivary nucleus. The projections from these three olivary areas were generally consistent with a zonal pattern of terminations in the FL and VP. The present results are consistent with a hypothesis that the FL is mainly involved in the control of vestibulo‐ocular reflex and that the VP is mainly involved in the control of smooth pursuit eye movements. J. Comp. Neurol. 382:480‐498, 1997.

Role of Primate Cerebellar Hemisphere in Voluntary Eye Movement Control Revealed by Lesion Effects

The anatomical connection between the frontal eye field and the cerebellar hemispheric lobule VII (H-VII) suggests a potential role of the hemisphere in voluntary eye movement control. To reveal the involvement of the hemisphere in smooth pursuit and saccade control, we made a unilateral lesion around H-VII and examined its effects in three Macaca fuscata that were trained to pursue visually a small target. To the step (3 degrees)-ramp (5-20 degrees/s) target motion, the monkeys usually showed an initial pursuit eye movement at a latency of 80-140 ms and a small catch-up saccade at 140-220 ms that was followed by a postsaccadic pursuit eye movement that roughly matched the ramp target velocity. After unilateral cerebellar hemispheric lesioning, the initial pursuit eye movements were impaired, and the velocities of the postsaccadic pursuit eye movements decreased. The onsets of 5 degrees visually guided saccades to the stationary target were delayed, and their amplitudes showed a tendency of increased trial-to-trial variability but never became hypo- or hypermetric. Similar tendencies were observed in the onsets and amplitudes of catch-up saccades. The adaptation of open-loop smooth pursuit velocity, tested by a step increase in target velocity for a brief period, was impaired. These lesion effects were recognized in all directions, particularly in the ipsiversive direction. A recovery was observed at 4 wk postlesion for some of these lesion effects. These results suggest that the cerebellar hemispheric region around lobule VII is involved in the control of smooth pursuit and saccadic eye movements.

Location of efferent terminals of the primate flocculus and ventral paraflocculus revealed by anterograde axonal transport methods.

Efferents of the flocculus (FL) and ventral paraflocculus (VP) were examined in seven anesthetized Macaca fuscata by anterograde axonal transport method using wheat germ agglutinin-conjugated horseradish peroxidase or phaseolus vulgaris leucoagglutinin. Several major foci of axon terminals were found in the vestibular nuclear complex and cerebellar nuclei. A difference was seen in the location of efferent terminals between the FL and VP. When the tracer covered the FL, labeled axon terminals were located within the medial and ventrolateral parts of the medial vestibular nucleus, superior vestibular nucleus and y-group. When the tracer covered the VP, labeled axon terminals were located within the caudo-ventral part of posterior interpositus and dentate nuclei, in addition to the medial and ventrolateral parts of the medial vestibular nucleus, superior vestibular nucleus and y-group. Labeled terminals were virtually absent in the basal interstitial nucleus of the cerebellum. On the points of neo- or paleo-cerebellar cortex fiber connections, these results correspond to our previous anatomical observations that the FL received mossy fiber afferents mainly from the vestibular system and nucleus reticularis tegmenti pontis and very little from the pontine nuclei, whereas the VP received mossy afferents mainly from the nucleus reticularis tegmenti pontis and pontine nuclei and very little from the vestibular system. These anatomical observations are consistent with a hypothesis in our previous anatomical and physiological study that the primate FL and VP mediate rather different functional roles in the oculomotor control.

Spinal cord cells innervating the bilateral parabrachial nuclei in the rat. A retrograde fluorescent double-labeling study.

The internal lateral nucleus (IL) of the parabrachial nucleus receives information from the spinal cord. The IL perhaps relays nociceptive signals to the intralaminar nuclei of the thalamus, apparently being implicated in the motivational-affective component of pain reactions. However, cells of origin of spinal fibers to the IL have not been investigated enough. We intended to clarify these cells, as well as their shapes, by retrograde double-labeling techniques. Fast blue and diamidino yellow dyes were injected, respectively, into the left and right ILs. The distribution of double-labeled cells was almost the same as that of single-labeled cells on both sides of the spinal cord. The total number of bilateral double-labeled cells was highest in the dorsolateral part of the lateral funiculus (DL), followed, in order, by lamina I, the dorsomedial part of the lateral funiculus (DM), lamina V and lamina VII. A few double-labeled cells were seen in laminae II-IV, VI, VIII and X. The ratio of the total number of bilateral double-labeled cells to the total number of bilateral single-labeled cells through the spinal cord was 43% in the DL, 37% in the DM, 28% in lamina V and 24% in lamina I. The ratio was 10% or less in the other remaining laminae. No marked differences were observed between the shapes of double- and single-labeled cells.

Role of primate cerebellar lobulus petrosus of paraflocculus in smooth pursuit eye movement control revealed by chemical lesion

The primate lobulus petrosus (LP) of the cerebellar paraflocculus receives inputs from visual system-related pontine nuclei, and projects to eye movement-related cerebellar nuclei. To reveal a potential involvement of LP in oculomotor control, we lesioned LP unilaterally by local injections of ibotenic acid in three Macaca fuscata. We examined the effects of lesion on eye movements evoked by step (3 degrees )-ramp (5-15 degrees/s) moving target. To step-ramp moving target, the monkeys showed an initial slow eye movement and later a small catch-up saccade, which was followed by the post-saccadic pursuit nearly matching to the velocity of the ramp target motion. After LP lesioning, the velocity of post-saccadic pursuits in the ipsiversive and down-ward directions decreased by 20-40% in all three monkeys. These deficits lasted for at least 1 month, and some recovery was observed. In the amplitudes of catch-up saccades, no consistent changes were seen among the three monkeys after LP lesioning. These results suggest an involvement of LP in the primate smooth pursuit eye movement control.

Changes in Expression of GABAA α4 Subunit mRNA in the Brain under Anesthesia Induced by Volatile and Intravenous Anesthetics

We investigated changes in levels of GABAA receptor α4 subunit mRNA in the mouse brain after administration of volatile or i.v. anesthetic, by performing quantitative RT-PCR. We also performed immunohistochemical assays for c-fos-like protein. During deep anesthesia (which was estimated by loss of righting reflex) after administration of propofol, levels of GABAA receptor α4 subunit mRNA in the hippocampus, striatum and diencephalons were significantly greater than those observed after administration of pentobarbital, midazolam or GOI (5.0% isoflurane and 70% nitrous oxide in oxygen). Under incomplete anesthesia, levels of GABAA receptor α4 subunit mRNA were significantly increased by midazolam in all brain regions, and were significantly increased by pentobarbital in the posterior cortex and striatum. Expression of GABAA receptor α4 subunit mRNA closely correlated with expression of c-fos-like protein. These results indicate that the GABAA receptor α4 subunit plays an important role in regulating the anesthetic stage of i.v. anesthetics.

Modulation of mThy28 nuclear protein expression during thymocyte development.

We have recently shown that down-regulation of mouse Thy28 (mThy28) protein expression appears to be accompanied by apoptotic processes. Thymocytes from mice contain moderate amounts of mThy28 protein and undergo proliferation, differentiation, or apoptosis during murine thymic maturation. As a first step to examine the potential role of the mThy28 protein in the thymocyte development, such as positive-negative selection, the expression of mThy28 protein in the thymocyte subsets was examined. Thymocytes are separated into four subpopulations by the expression levels of CD4 and CD8: CD4-CD8- (DN), CD4+CD8+ (DP), and CD4+CD8- or CD4-CD8+ (SP). Flow cytometry analysis using three-color staining demonstrated that the mThy28 expression in immature DP cells is lower than that in DN and SP cells. The down-regulation of the mThy28 expression in the DP stage was also detected by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). The immunostaining method also showed that mThy28 protein was expressed in the medulla containing mature thymocytes, but not the cortex having immature thymocytes. The mThy28 protein in the thymocytes was mainly localized in the nucleus, as recently demonstrated in lymphoma cells, indicating that the mThy28 protein resides in the nucleus, irrespective of the cyclic or resting stage of the cell cycle. Together, the observation that mThy28 expression is down-modulated during the DP stage suggests that mThy28 protein might play some role in the positive-negative selection step in thymic maturation.

Cytoarchitectonic subdivisions of the parabrachial nucleus in the Japanese monkey (Macacus fuscatus) with special reference to spinoparabrachial fiber terminals

The cytoarchitectonic subnuclear organization of the parabrachial nucleus (PB) surrounding the brachium conjunctivum (BC) in the monkey was examined using the Nissl method and the anterograde axonal flow method. PB of the monkey could be divided into the following subnuclei: the dorsal area (DPBM) along the medial surface of the medial three-fourths of BC in the caudal half of medial PB (PBM), the ventral area (VPBM) along the medial surface of the lateral one-fourth of BC in the rostral two-thirds of PB, the ventrolateral part of lateral PB (PBL) lateral to BC throughout PB (EL), the ventral part of the rostral half of PBL ventral to EL (EXL), the medial part of middle PBL along the dorsal surface of BC (VL), the dorsal and lateral marginal part of PBL in the rostral two-thirds of PB (DL), the cell cluster in the dorsomedial part of the rostral half of PBL between VL and DL (CL), the dorsocentral part appearing at the level of root exit of the trochlear nerve between DL and CL and extending to the rostral end of PBL (IL), the area between DL and IL in the rostral one-seventh of PBL (SL), and Kölliker-Fuse nucleus (KF) ventral to EL and BC in the middle one-third of PB and lateral to the lateral pontine tegmentum. After the injection of biotinylated dextran amine into the upper cervical segments, labeled fibers terminated in each subdivision of PB with different densities; most heavily in IL, more heavily in DL and KF, moderately in EL and VPBM, and scarcely in the rest of PB. The present study demonstrated for the first time the subdivisions of PB in the monkey, which were essentially common to those of the rat based on the cytoarchictecture of PB and spinal fiber terminals in it.

Morphological changes indicative of brain hemisphere development and handedness in fixed brains and on CT/MRI images

We measured the lengths of some parts of the right and left hemispheres (HEs) in 70 formalin-fixed brains and on 15 computed tomography/magnetic resonance imaging (CT/MRI) images (7 left-handed and 8 right-handed cases) to clarify the morphological changes indicating which HE developed earlier and handedness. In many cases of the fixed brains, 1) the distance from the frontal pole to the occipital pole was longer in the left HE than in the right HE, 2) the distance from the middle plane to the lateral-most portion of the HE was wider in the right HE than in the left HE, 3) the left occipital pole elongated more posteriorly and covered the right occipital pole, and 4) the volume of each HE was nearly the same. The results indicate that the left HE develops and grows slightly earlier in the larger semi-cranium (half of the cranium) than the right HE which develops later in the smaller semi-cranium. The whole brain was more spherical in the female cases than in the male cases. The morphological changes in both HEs for handedness were not evident on the CT/MRI images.

Three-dimensional structures of the dissected human brain: Visible with naked eye

We analyzed the brain activity related to the performance of the Wisconsin Card Sorting Test (WCST) by using functional magnetic resonance imaging (fMR1) in seven human subjects. In our computerized WCST, subjects continually sorted card stimuli using their right hands throughout runs. and. in addition, intermittently shifted cognitive sets after dimensional changes. Therefore, total MR signals observed during performance of the WCST contained both transient and sustained signals. Transient signals were elicited by the shifting of cognitive sets, and sustained signals were elicited by the sorting of card stimuli. We previously isolated the shift-related transient MR signals by comparing the MR signals obtained after the dimensional changes with those obtained before the dimensional changes, and subtracting out the baseline activation level derived from the sorting of card stimuli (S. Konishi et al., Nature Neurosci 1998). In the present study, we report on the baseline activation. We compared the MR signals obtained during sorting with those obtained during fixation. Activation was found mainly in the visual cortex, the left primary motor and two premotor areas. But these areas did not overlap with the shift-related areas, indicating that the validity of the decomposition into the shift-related and sort-related areas.