Of the six glutamate receptor (GluR) channel subunit families identified by molecular cloning, five have been shown to constitute either the AMPA, kainate, or NMDA receptor channel, whereas the function of the delta subunit family remains unknown. The selective localization of the delta 2 subunit of the GluR delta subfamily in cerebellar Purkinje cells prompted us to examine its possible physiological roles by the gene targeting technique. Analyses of the GluR delta 2 mutant mice reveal that the delta 2 subunit plays important roles in motor coordination, formation of parallel fiber-Purkinje cell synapses and climbing fiber-Purkinje cell synapses, and long-term depression of parallel fiber-Purkinje cell synaptic transmission. These results suggest a close relationship between synaptic plasticity and synapse formation in the cerebellum.

Conjunctive stimulation of climbing and parallel fibres in the cerebellum evokes a long-term depression of parallel-fibre Purkinje-cell transmission, a phenomenon implicated as the cellular mechanism for cerebellar motor learning. It is suspected that the increase in cyclic GMP concentration that occurs after activation of climbing fibres is required to evoke long-term depression. Excitatory amino acids are known to cause the release of nitric oxide (NO), resulting in elevation of the cGMP level in the cerebellum. Here we report that endogenous NO is released after stimulation of climbing fibres, that long-term depression evoked by conjunctive stimulation of parallel and climbing fibres is blocked by haemoglobin (which strongly binds NO) or L-NG-monomethyl-arginine (an inhibitor of NO synthase), and that exogenous NO or cGMP can substitute for the stimulation of climbing fibres to cause long-term depression in rat cerebellar slices. These results demonstrate that the release of endogenous NO is essential for the induction of synaptic plasticity in the cerebellum.

Mice devoid of glial fibrillary acidic protein (GFAP), an intermediate filament protein specifically expressed in astrocytes, develop normally and do not show any detectable abnormalities in the anatomy of the brain. In the cerebellum, excitatory synaptic transmission from parallel fibers (PFs) or climbing fibers (CFs) to Purkinje cells is unaltered, and these synapses display normal short-term synaptic plasticity to paired stimuli in GFAP mutant mice. In contrast, long-term depression (LTD) at PF-Purkinje cell synapses is clearly deficient. Furthermore, GFAP mutant mice exhibited a significant impairment of eyeblink conditioning without any detectable deficits in motor coordination tasks. These results suggest that GFAP is required for communications between Bergmann glia and Purkinje cells during LTD induction and maintenance. The data support the notion that cerebellar LTD is a cellular mechanism closely associated with eyeblink conditioning, but is not essential for motor coordination tasks tested.

We used autofluorescence of mitochondrial flavoproteins to image cortical neural activity in the rat. Green autofluorescence in blue light was examined in slices obtained from rat cerebral cortex. About half of the basal autofluorescence was modulated by the presence or absence of O2 or glucose in the medium. Repetitive electrical stimulation at 20 Hz for 1 s produced a localized fluorescence increase in the slices. The amplitude of the increase was 27 +/- 2 % (mean +/- S.D., n = 35). Tetrodotoxin or diphenyleneiodonium, an inhibitor of flavoproteins, blocked the autofluorescence responses. The autofluorescence responses were not observed in slices perfused with calcium-, glucose- or O2-free medium. In the primary somatosensory cortex of rats anaesthetized with urethane (1.5 g kg-1, I.P.), an activity-dependent increase in autofluorescence of 20 +/- 4 % (n = 6) was observed after electrical cortical stimulation at 100 Hz for 1 s, and an increase of 2.6 +/- 0.5 % (n = 33) after vibratory skin stimulation at 50 Hz for 1 s applied to the plantar hindpaw. These responses were large enough to allow visualization of the neural activity without having to average a number of trials. The distribution of the fluorescence responses after electrical or vibratory skin stimulation was comparable to that of the cortical field potentials in the same rats. The fluorescence responses were followed by an increase in arterial blood flow. The former were resistant to an inhibitor of nitric oxide synthase, while the latter was inhibited. Thus, activity-dependent changes in the autofluorescence of flavoproteins are useful for functional brain imaging in vivo.

The preservation of visual property differences among the higher visual areas following V1 lesions and their loss following SC lesions indicate that pathways from the SC through the thalamus to higher cortical areas are sufficient to support these differences.

No abstract

The primary auditory cortex (AI) is the representative recipient of information from the ears in the mammalian cortex. However, the delineation of the AI is still controversial in a mouse. Recently, it was reported, using optical imaging, that two distinct areas of the AI, located ventrally and dorsally, are activated by high-frequency tones, whereas only one area is activated by low-frequency tones. Here, we show that the dorsal high-frequency area is an independent region that is separated from the rest of the AI. We could visualize the two distinct high-frequency areas using flavoprotein fluorescence imaging, as reported previously. SMI-32 immunolabeling revealed that the dorsal region had a different cytoarchitectural pattern from the rest of the AI. Specifically, the ratio of SMI-32-positive pyramidal neurons to nonpyramidal neurons was larger in the dorsal high-frequency area than the rest of the AI. We named this new region the dorsomedial field (DM). Retrograde tracing showed that neurons projecting to the DM were localized in the rostral part of the ventral division of the medial geniculate body with a distinct frequency organization, where few neurons projected to the AI. Furthermore, the responses of the DM to ultrasonic courtship songs presented by males were significantly greater in females than in males; in contrast, there was no sex difference in response to artificial pure tones. Our findings offer a basic outline on the processing of ultrasonic vocal information on the basis of the precisely subdivided, multiple frequency-organized auditory cortex map in mice.

1. Nitric oxide (NO) release following repetitive electrical stimulation was studied in the molecular layer of rat cerebellar slices using electrochemical NO probes. 2. In parasagittal slices of the vermis, most Purkinje cells showed climbing fibre responses in response to white matter stimulation without accompanying NO release. 3. In frontal slices, parallel fibre volley potentials and NO release were elicited concurrently by parallel fibre stimulation. 4. The NO release following parallel fibre stimulation was not affected by blockers of non-NMDA, NMDA and metabotropic glutamate receptors. 5. The NO release was reduced significantly (P < 0f001) to 29% of the control level after climbing fibre deafferentation with 3-acetylpyridine treatment.6. The rate of NO release was roughly proportional to the second or third power of the stimulus frequency, and to the third power of the extracellular Ca2+ concentration. 7. The rate of NO release was not affected by nicardipine (10 ASM). It was reduced to 87 + 4%(n = 5, mean + S.E.M.) of the control level by w-conotoxin GVIA (0 3 uM), and to 18 + 4% (n = 4) by w-agatoxin IVA (0 3 uM). 8. Tetanic parallel fibre stimulation potentiated NO release by 24 + 5 % (n = 5). 9. These data indicate that NO is derived mainly from parallel fibres. The relationship between NO release and cerebellar synaptic plasticity is discussed.Of the isoforms of nitric oxide (NO) synthase (NOS), the neuronal type (nNOS) is distributed widely in the brain (Bredt & Snyder, 1994). The activity of nNOS is modulated by calcium through calmodulin, and the product of nNOS, NO, can diffuse through the cell membrane freely. NO may therefore be an intercellular messenger that carries information without depending on synaptic connections. However, the exact function of NO in the brain is not very clear. Among various functions assigned to NO, the role in synaptic plasticity is probably the most enigmatic. NO synthesis in the brain was first detected in the cerebellum (Garthwaite, Charles & Chess-Williams, 1988), where nNOS is present at a high concentration (Bredt, Hwang & Snyder, 1990) and most of the NOS activity is eliminated by disruption of the nNOS gene (Huang, Dawson, Bredt, Snyder & Fishman, 1993). The main synaptic plasticity in the cerebellum is manifested by long-term depression (LTD) in parallel fibre-Purkinje cell synapses, which is suggested to be the cellular mechanism for motor learning (Ito, 1989 (Ito, Sakurai & Tongroach, 1982;Sakurai, 1987). In a study using caged NO, LTD could be induced by light-liberated NO as well as by parallel fibre stimulation (Lev-Ram, Makings, Keitz, Kao & Tsien, 1995), strongly suggesting that NO is released from parallel fibres in order to induce LTD. The purpose of the present study was to demonstrate and characterize possible NO release from parallel fibres.In our previous studies, NO release was demonstrated in the molecular layer of cerebellar slices following white matter stimulation (Shibuki, 1990b;Shibuki & Okada, 1991