Frans VanderWerf

Deletion of FMR1 in Purkinje cells enhances parallel fiber LTD, enlarges spines, and attenuates cerebellar eyelid conditioning in Fragile X syndrome.

Absence of functional FMRP causes Fragile X syndrome. Abnormalities in synaptic processes in the cerebral cortex and hippocampus contribute to cognitive deficits in Fragile X patients. So far, the potential roles of cerebellar deficits have not been investigated. Here, we demonstrate that both global and Purkinje cell-specific knockouts of Fmr1 show deficits in classical delay eye-blink conditioning in that the percentage of conditioned responses as well as their peak amplitude and peak velocity are reduced. Purkinje cells of these mice show elongated spines and enhanced LTD induction at the parallel fiber synapses that innervate these spines. Moreover, Fragile X patients display the same cerebellar deficits in eye-blink conditioning as the mutant mice. These data indicate that a lack of FMRP leads to cerebellar deficits at both the cellular and behavioral levels and raise the possibility that cerebellar dysfunctions can contribute to motor learning deficits in Fragile X patients.

Eyelid movements during blinking in patients with Parkinson's disease

We examined eyelid movements during spontaneous, voluntary, and trigeminal reflex blinks in 16 patients with mild to moderate Parkinsons disease (PD) off medication and 14 controls. Voluntary and reflex blink amplitudes tended to be smaller than normal for PD patients, whereas eyelid kinematics (amplitude–maximum velocity relationship) for all three blink types were normal. Spontaneous blink rate was less than normal for 10 patients and abnormally high for 6 patients. A significant positive correlation between spontaneous blink amplitude and blink rate was found. These observations suggest that PD modifies the gain of a premotor blink circuit shared by spontaneous, voluntary, and reflex blinks.

Anti-malaria drug mefloquine induces motor learning deficits in humans

Mefloquine (a marketed anti-malaria drug) prophylaxis has a high risk of causing adverse events. Interestingly, animal studies have shown that mefloquine imposes a major deficit in motor learning skills by affecting the connexin 36 gap junctions of the inferior olive. We were therefore interested in assessing whether mefloquine might induce similar effects in humans. The main aim of this study was to investigate the effect of mefloquine on olivary-related motor performance and motor learning tasks in humans. We subjected nine participants to voluntary motor timing (dart throwing task), perceptual timing (rhythm perceptual task) and reflex timing tasks (eye-blink task) before and 24 h after the intake of mefloquine. The influence of mefloquine on motor learning was assessed by subjecting participants with and without mefloquine intake (controls: n = 11 vs mefloquine: n = 8) to an eye-blink conditioning task. Voluntary motor performance, perceptual timing, and reflex blinking were not affected by mefloquine use. However, the influence of mefloquine on motor learning was substantial; both learning speed as well as learning capacity was impaired by mefloquine use. Our data suggest that mefloquine disturbs motor learning skills. This adverse effect can have clinical as well as social clinical implications for mefloquine users. Therefore, this side-effect of mefloquine should be further investigated and recognized by clinicians.

Savings and extinction of conditioned eyeblink responses in fragile X syndrome

The fragile X syndrome (FRAXA) is the most widespread heritable form of mental retardation caused by the lack of expression of the fragile X mental retardation protein (FMRP). This lack has been related to deficits in cerebellum‐mediated acquisition of conditioned eyelid responses in individuals with FRAXA. In the present behavioral study, long‐term effects of deficiency of FMRP were investigated by examining the acquisition, savings and extinction of delay eyeblink conditioning in male individuals with FRAXA. In the acquisition experiment, subjects with FRAXA displayed a significantly poor performance compared with controls. In the savings experiment performed at least 6 months later, subjects with FRAXA and controls showed similar levels of savings of conditioned responses. Subsequently, extinction was faster in subjects with FRAXA than in controls. These findings confirm that absence of the FMRP affects cerebellar motor learning. The normal performance in the savings experiment and aberrant performance in the acquisition and extinction experiments of individuals with FRAXA suggest that different mechanisms underlie acquisition, savings and extinction of cerebellar motor learning.

Long-term changes in cerebellar activation during functional recovery from transient peripheral motor paralysis.

Localized altered cerebellar cortical activity can be associated with short-term changes in motor learning that take place in the course of hours, but it is unknown whether it can be correlated to long-term recovery from transient peripheral motor diseases, and if so, whether it occurs concomitantly in related brain regions. Here we show in a longitudinal fMRI study of patients with unilateral Bells palsy that increases in ipsilateral cerebellar activity follow the recovery course of facial motor functions over at least one and a half years. These findings hold true for changes in brain activity related to both oral and peri-orbital activation, even though these processes are differentially mediated by unilateral and bilateral brain connectivities, respectively. Activation of non-facial musculature, which was studied for control, does not show any change in cerebellar activity over time. The localized changes in cerebellar activities following activation of facial functions occur concomitantly with increases in activity of the facial region in the contralateral primary motor cortex suggesting that the cerebellum acts together with the cerebral cortex in long-term adaptation to transient pathological sensorimotor processing.

Exceptional good cognitive and phenotypic profile in a male carrying a mosaic mutation in the FMR1 gene.

Fragile X (FRAX) syndrome is a commonly inherited form of mental retardation resulting from the lack of expression of the fragile X mental retardation protein (FMRP). It is caused by a stretch of CGG repeats within the fragile X gene, which can be unstable in length as it is transmitted from generation to generation. Once the repeat exceeds a threshold length, the FMR1 gene is methylated and no protein is produced resulting in the fragile X phenotype. The consequences of FMRP absence in the mechanisms underlying mental retardation are unknown. We have identified a male patient in a classical FRAX family without the characteristic FRAX phenotype. His intelligence quotient (IQ) is borderline normal despite the presence of a mosaic pattern of a pre‐mutation (25%), full mutation (60%) and a deletion (15%) in the FMR1 gene. The cognitive performance was determined at the age of 28 by the Raven test and his IQ was 81. However, FMRP expression studies in both hair roots and lymphocytes, determined at the same time as the IQ test, were within the affected male range. The percentage of conditioned responses after delay eyeblink conditioning was much higher than the average percentage measured in FRAX studies. Moreover, this patient showed no correlation between FMRP expression and phenotype and no correlation between DNA diagnostics and phenotype.

Reticulo-collicular and spino-collicular projections involved in eye and eyelid movements during the blink reflex

Reflex blinking provides a useful experimental tool for various functional studies on the peripheral and central nervous system, yet the neuronal circuitry underlying this reflex is not precisely known. In the present study, we investigated as to whether neurons in the reticular formation and rostral cervical spinal cord (C1) may be involved in the blink reflex in rats. To this end we investigated c-Fos expression in these areas following supraorbital nerve stimulation combined with retrograde tracing of gold conjugated horse radish peroxidase (Gold-HRP) from the superior colliculus. We observed many double labeled neurons in the parvocellular reticular nucleus, medullary reticular formation, and laminae IV and V of C1. Thus, these brain regions contain neurons that may be involved in blink reflexes as well as eye movements, because they both can be activated following peri-orbital stimulation and project to the superior colliculus. Consequently, we suggest that the medullary reticular formation and C1 region play a central role in the coordination of eye and eyelid movements during reflex blinking.

Are locus coeruleus neurons involved in blinking

To investigate the involvement of the noradrenergic locus coeruleus (LC) in the reflex blink circuit, c-Fos and neuronal tracer experiments were performed in the rat. LC neurons involved in reflex blink were localized by analyzing c-Fos protein expression after electrical stimulation of the supraorbital nerve. Subsequently, neuronal tracers were injected in two different nuclei which are part of the reflex blink circuit. Anterograde tracer experiments in the sensory trigeminal complex (STC) explored the trigemino-coerulear connection; retrograde tracer experiments in the latero-caudal portion of the superior colliculus (SC) established coerulear-collicular connections. The combination of retrograde tracer injections into the latero-caudal SC portion combined with electrical stimulation of the supraorbital nerve identified c-Fos positive LC neurons that project to the latero-caudal SC. Our results revealed the existence of a STC-LC-SC loop.

Reticulo-collicular projections: a neuronal tracing study in the rat

Neuroanatomical tract-tracing methods were used to study the topography of the reticulocollicular projections. Injections of gold-HRP or BDA tracers into the medial and/or central portions of the superior colliculus resulted in labelled neurones mainly in the medial reticular formation, whereas injections into the lateral portion of the superior colliculus showed labelling in the medial and lateral reticular formation. When tracer was injected into the lateral portion of the caudal superior colliculus, extensive lateral labelling was observed in the contralateral parvocellular reticular nucleus and the contralateral dorsal medullary reticular nucleus, two areas involved in reflex blinking. The present study shows that these reticular areas project to the lateral superior colliculus, which is known to be involved in the coordination of eye and eyelid movements.

Connections between the lacrimal gland and sensory trigeminal neurons: a WGA/HRP study in the cynomolgous monkey

The sensory innervation of the lacrimal gland (LG) in the cynomolgous monkey was studied using the retrograde wheat germ agglutinin/horsereadish peroxidase (WGA/HRP) tracer technique. A small solidified piece of WGA/HRP was implanted in the LG. Labelled sensory first‐order neurons were found in the ipsilateral trigeminal ganglion (TG) and in the ipsilateral mesencephalic trigeminal nucleus (MTN). The distribution of labelled TG neurons was restricted to ophthalmic and maxillary ganglionic parts. Sensory innervation of LG by primary afferents is not only restricted to TG; an MTN involvement has also been found. This may imply that there is a central sensory role in the production and release of tears.