Inhibitory influences on receptive field size in the dorsal column nuclei.

Receptive fields (RFs) of neurons in the dorsal column nuclei (DCN) expand within minutes after the RFs are anesthetized via subcutaneous lidocaine injections (e.g., Pettit and Schwark). The mechanism of this rapid reorganization is of great interest. It has been proposed that such RF expansion results from a decline in inhibition within the DCN that unmasks previously ineffective synapses. To study the role of GABAergic inhibition in the DCN in controlling RF size, we applied by iontophoresis bicuculline methiodide to block gamma-aminobutyric acidA (GABA(A)) receptors and 2-OH-saclofen to block GABA(B) receptors. Blockade of GABA(A) receptors resulted in RF expansions in 79% of the neurons, while blockade of GABA(B) receptors resulted in RF expansions in 53% of the neurons. The effectiveness of receptor blockade in producing RF expansion was not related to neuronal response characteristics. Glutamate application resulted in RF expansions in only 2 of 23 neurons tested, suggesting that RF expansion was not simply due to increased excitability. The modality and adaptation characteristics of the expanded portions of the RFs were similar to those of the original RF. The results of the present study suggest that GABAergic inhibition can play a role in controlling RF size in the DCN, and that both GABA(A) and GABA(B) receptors may be involved in this process.

Distribution of substance P receptor binding in dorsal column nuclei of rat, cat, monkey and human.

In the present study, substance P receptor binding was localized in the dorsal column nuclei (DCN) of the rat, cat, monkey, and human. Bolton-Hunter-labeled [125I]substance P binding was most concentrated in the cell nests of the core region, but was present throughout the DCN of each species. The distribution of substance P receptors may reconcile apparent mismatches between the widespread responsiveness of DCN neurons to substance P and the restricted distribution of substance P containing afferents.

Effects of whisker trimming on GABA(A) receptor binding in the barrel cortex of developing and adult rats.

Both sensory deprivation and blockade of gamma-aminobutyric acid A (GABA(A)) receptors result in signs of cortical disinhibition. To investigate whether down-regulation of GABA(A) receptors could underlie effects of sensory deprivation, [3H]muscimol binding was assessed in rat whisker barrels after chronic whisker trimming. Vibrissae in row C or rows A,B,D, and E were trimmed during certain developmental periods. When whiskers were trimmed for the first 6 postnatal weeks, [3H]muscimol binding was 8.3% lower in deprived barrel rows than in adjacent nondeprived rows (P < 0.001). The effect may be somewhat selective for GABA(A) receptors because there was no evident change in N-methyl-D-aspartate (NMDA) receptors as indicated by [3H]MK-801 binding. Ten weeks after whiskers were allowed to regrow, the decrease in [3H]muscimol binding was partly reversed (P < 0.002), leaving a 3.3% decrease (P < 0.001). These declines in GABA(A) receptors could contribute to persisting electrophysiological signs of reduced inhibition in similarly deprived barrel neurons (Simons and Land [1987] Nature 326:694-697). A 6-week deprivation beginning in adulthood resulted in a 7.7% decrease (P < 0.001), indicating that the effect is not restricted to an early critical period. In rats trimmed for the first 10 postnatal days, [3H]muscimol binding declined 2.3% (P < 0.05), which is a small change compared with the magnitude of the developmental peak; thus, normal whisker input apparently is not required for the developmental increase in GABA(A) receptors. The present study suggests that sensory input can regulate cortical GABA(A) receptors in adulthood and during ontogeny. Down-regulation of cortical GABA(A) receptors may be a compensatory mechanism that serves to disinhibit the reduced sensory input.

Regional distribution of GABAA receptor binding sites in cat somatosensory and motor cortex.

Inhibition in primary sensory cortex plays a role in neuronal responses to peripheral stimuli. For many neurons in cat primary somatosensory cortex, blockade of GABAA receptors by bicuculline results in receptive field enlargement. The magnitude of this effect varies with the neuron's adaptation characteristics and its location in particular laminae and submodality regions. To test whether these variations are correlated with the distribution of GABAA receptors, we analyzed [3H]muscimol binding in cat primary somatosensory and motor cortical areas. The highest levels of binding were in layers I-III, and the lowest levels were in layers V-VI. In somatosensory cortical areas, layer IV was distinguished by higher levels of binding than in adjacent layers. Within layer IV, levels of binding were significantly higher in posterior area 3b than in anterior area 3b. These differences may correspond to the rapidly adapting and slowly adapting submodality regions which have been described in this area. The laminar distribution of [3H]muscimol binding differed from that of [3H]flunitrazepam, and neither resembled the distribution of the magnitude of bicuculline's effects on receptive field size. The laminar distribution of [3H]muscimol binding was highly correlated with the areal density of GABA-immunoreactive neurons described in a companion study.

NMDA receptor binding in rodent suprachiasmatic nucleus.

NMDA receptors are thought to mediate effects of light on circadian rhythms and on immediate-early gene expression in the suprachiasmatic nucleus (SCN), the primary circadian pacemaker in mammals. The present study characterized NMDA receptors in autoradiographs of SCN incubated with the NMDA antagonist [3H]MK-801. In both rat and hamster, [3H]MK-801 binding did not delineate the SCN and was fairly uniformly distributed across the SCN region. Binding levels were unaffected by circadian time, light vs. dark conditions, or enucleation. Scatchard analyses revealed species differences in both receptor number and affinity in the SCN. The [3H]MK-801 binding sites characterized in this study could mediate the NMDA antagonist-sensitive effects of light on the SCN and circadian rhythms.

Distribution of [3H]QNB and [125I]alpha-bungarotoxin binding and acetylcholinesterase activity in visual system and hippocampal structures of eleven mammalian species.

This study assessed interspecies differences in regional brain distribution of [3H]QNB binding, [125I]alpha-bungarotoxin binding and acetylcholinesterase activity, by autoradiographic and histochemical methods. Eleven mammalian species were examined, including carnivores (cat, dog), a lagomorph (rabbit), and rodents (squirrel, guinea pig, gerbil, hamster, vole, lemming, rat, mouse). Comparisons were based on primary visual system structures (superior colliculus, lateral geniculate nucleus, primary visual cortex) and the hippocampal formation. The two radioligands differed greatly in the degree of interspecies variation: while the pattern of [3H]QNB binding was quite similar across species, [125I]alpha-bungarotoxin showed striking interspecies diversity. This contrast was most obvious in laminar patterns of the visual cortex and hippocampal formation. Regional distributions of acetylcholinesterase staining were fairly diverse, and were unlike the patterns of either [3H]QNB or [125I]alpha-bungarotoxin. The two ligands showed more consistency in overall levels across species than did acetylcholinesterase. Possible correlates of the differences in interspecies diversity are discussed.

Distribution of NMDA receptor binding in developing mouse spinal cord.

The role of NMDA receptors in spinal sensorimotor processes has been studied in neonates and adults, while NMDA binding has previously been described only in mature spinal cord. In autoradiographs of mouse lumbar cord, NMDA-sensitive [3H]glutamate binding peaked transiently around postnatal days 6-10. Differentiation increased progressively from day 10 to adulthood. Mature levels were highest in substantia gelatinosa and moderate in dorsomedial dorsal horn. The results are described in relation to functional and anatomical features of developing spinal cord.

[125I]Vasoactive intestinal peptide binding in rodent suprachiasmatic nucleus: developmental and circadian studies.

The suprachiasmatic nucleus (SCN) of rat and hamster have been studied extensively and shown to play critical roles in circadian rhythmicity. [125I]Vasoactive intestinal peptide (VIP) binding levels are high in the rat SCN, suggesting that VIP receptors may be an important component of SCN function. In contrast to previously demonstrated diurnal variations in VIP immunoreactivity and VIP mRNA, the present study found [125I]VIP binding to be stable across the light-dark cycle in both rat and hamster SCN. High [125I]VIP labeling appeared to be coextensive with the rat SCN but extended somewhat beyond the cytoarchitectonic boundaries of the hamster SCN. Binding density in hamster SCN was slightly higher than in rat. In the developing rat SCN, [125I]VIP binding levels distinguished the SCN on embryonic day 18, and appeared to increase to postnatal day 10 before declining to adult levels. The early presence of [125I]VIP binding suggests possible involvement of VIP receptors in fetal entrainment of circadian rhythms.

5'-Nucleotidase activity increases in aging rat brain.

The enzyme 5'-nucleotidase is present in glial and neuronal membranes, and catalyzes the formation of adenosine, which in turn can act as a neuromodulator or neurotransmitter. The present study found marked increases in histochemically demonstrated 5'-nucleotidase activity in most regions of rat brain from young adulthood (3-4 months) to middle age (12-18 months), with smaller or no changes between middle and old age (24-32 months). The aging adult cerebellum showed alterations in the histochemical pattern, with declines in the molecular layer and increases in the Purkinje layer. Both myelin and synaptic plasma membrane fractions from forebrain showed increases in enzyme activity. Assays of various other body tissues suggested that the increases are fairly specific to brain, and thus apparently do not represent a ubiquitous cellular mechanism of aging. Changes in brain 5'-nucleotidase activity during aging probably reflect the increasing number and size of glial cells, and perhaps also affect synaptic transmission through regulation of adenosine.

Effect of environmental complexity on size of the superior colliculus.

The present study examined effects of environmental complexity on the size of the superior colliculus, a subcortical structure involved in visuomotor functions. Long-Evans hooded rats raised together in a complex environment for 48 weeks were compared with their littermates housed in individual cages. The depth and area of the superficial gray layer of the superior colliculus were about 5-6% greater in the group from the complex environment, while the deeper layers of the superior colliculus showed no significant differences. The magnitude of the differences approached those reported for the neocortex, which has been considered to be distinctive in its morphological responsiveness to differential environmental complexity. The findings also extend previous observations that visual deprivation leads to shrinkage of the superficial gray layer and indicate that the morphology of this subcortical visual area is responsive to varying degrees of environmental stimulation.

[125I]alpha-bungarotoxin binding marks primary sensory area developing rat neocortex.

The postnatal ontogeny of [125I]alpha-bungarotoxin (alpha-Btx) binding distribution in rat neocortex was described and quantified using autoradiography of in vitro labeled brain sections. During the first two weeks, distinctive transitory radial and laminar patterns emerged. Dense columnar bands of alpha-Btx binding extended through the depth of primary sensory cortex, including somatosensory, visual and auditory areas. An association of alpha-Btx binding with thalamic input zones was further demonstrated within developing somatosensory cortex, where discrete radial bands appeared over the whisker barrels around the time that ingrowing thalamocortical fibers segregate as they selectively innervate the barrels. The early laminar distribution of alpha-Btx binding also resembled that of developing thalamocortical afferents. From P12 to P20, alpha-Btx radial distinctions faded and the laminar pattern changed further to achieve the adult distribution. The spatiotemporal ontogeny of alpha-Btx binding suggests a role for alpha-Btx binding sites in the development of cortical connectivity.

Alpha-bungarotoxin binding in relation to functional organization of the rat suprachiasmatic nucleus.

[125I]alpha-Bungarotoxin (alpha-Btx) binding was quantified in autoradiographs of in vitro labeled sections through the rat suprachiasmatic nucleus (SCN). Recent evidence suggests that acetylcholine and alpha-Btx binding sites in the SCN region may mediate effects of light on circadian functions. In contrast to the ventrolateral termination of retinal axons, there was a consistent pattern of relatively high alpha-Btx binding levels in the dorsolateral SCN and low levels in the ventromedial SCN. In addition, enucleation had little or no effect. We also investigated the possibility that phase-dependent influences of light on circadian functions could reflect the presence of light-induced or daily rhythms in alpha-Btx binding. Animals were tested in a light-dark cycle or after 1-2 days in constant light. The dorsolateral SCN showed a small but significant decline of alpha-Btx binding in animals in light at night compared to darkness at night. alpha-Btx binding in the SCN showed no statistically significant daily variation in animals in the light-dark cycle.

Two-point discriminability: relation to properties of the somatosensory system.

The ability to resolve two closely spaced cutaneous stimuli presumably depends upon the degree of overlap between the two populations of responding neurons. The degree of overlap is determined by receptive field (RF) geometry and location, and by interactive factors such as lateral inhibition. In this paper, we first consider some aspects of RFs that would be expected to influence two-point acuity. In some somatosensory brain regions, relatively few RFs overlap the body midline. As would be expected, discrimination is enhanced for two points straddling the backbone. This does not simply reflect a mediolateral gradient of acuity, as we found higher acuity laterally. On the limbs, where RFs are elongated along the length of the limb, transverse two-point acuity was greater than longitudinal acuity. However, on the back, where RFs are fairly round, there was an even larger orientation effect, with two-point acuity greater for stimuli longitudinal than for stimuli transverse to the spine. Thus, the substantial variation of two-point acuity with stimulus orientation on the back cannot be explained by RF geometry alone. We discuss the possibility that differences in lateral inhibition and degree of similarity in dermatomal composition contribute to the observed stimulus orientation effects.

Development of circadian rhythmicity and light responsiveness in the rat suprachiasmatic nucleus: a study using the 2-deoxy[1-14C]glucose method.

The suprachiasmatic nucleus (SCN) of the hypothalamus is thought to play a critical role in circadian rhythm generation and entrainment to the light/dark cycle. In adult rats, the SCN shows a circadian rhythm in metabolic activity level as indicated by 2-deoxy[1-14C]glucose uptake. In the present study, the development of this rhythm was investigated. No diurnal difference in uptake was evident in fetal rats 1--2 days before birth. A significant diurnal difference in SCN 2-deoxyglucose uptake was preseno light at night increased the SCN metabolic levels. According to previous studies, on day 1 the SCN is poorly developed and contains few synapses. At this time the retinohypothalamic tract has not yet developed. We found progressive functional maturation of the SCN through day 21, when the rhythm and light responsiveness resembled those of adult rats.

Somatotopic representation of hindlimb skin in cat dorsal horn.

Single-unit exploration of the dorsal horn of segments L4-S2 of unanesthetized cats with the neuraxis transected at lower thoracic levels reveals a somototopic organization in the horizontal plane. The dorsal horn dermatomes correspond closely to the dermatomes of the corresponding dorsal roots, and the ML gradient is equally well described by two different projection schemes: a distoproximal gradient and a ventrodorsal one (5, 33). There is no evidence of segmental discontinuity of the map. As is the case in other nuclear regions of the CNS, the relative area devoted to projections from the foot is disproportionately large relative to the area devoted to skin regions of similar size which are located more proximally on the limb. From our data, and from the close correspondence to anatomical data obtained by others, we suggest that at least some cutaneous afferent fibers from a given skin area project directly to any dorsal horn region where that skin area is represented. This assumption, together with the organization of the dorsal horn map, yields a model which predicts a precise somatotopic organization of presynaptic neuropil in the substantia gelatinosa.

Parametric studies of dorsal horn neurons responding to tactile stimulation.

Dorsal horn neurons responding to tactile input were recorded in segments L3-S2 of unanesthetized, low-spinal cats. Single units were characterized with regard to receptive field (RF) location, RF size and shape, spontaneous discharge rate, central delay, and convergence of four tactile afferent types. 1. RF size increased from the toes to the calf region. 2. Length-width ratio increased from the toes to the calf and declined from the calf to the hip. 3. The relation between RF size and position on the limb was independent of segmental and laminar location of the neurons. 4. RF size was positively correlated with spontaneous discharge rate. 5. The relation between RF size and shape and RF position can be interpreted in terms of regional variations in the magnitude of the gradient of representation in the dorsal horn somatotopic map. 6. Central delay was negatively correlated with both RF size and rate of ongoing discharge. 7. There were no statistically significant differences among the laminae with respect to central delay, RF size or shape, ongoing discharge, or convergence combinations of the four tactile afferent systems. 8. Data presented were at variance with Wall's laminar cascading model for laminae IV-VI. Our results suggest that the model should be modified, at least to emphasize monosynaptic tactile input to all three of these laminae.