Case series evidence for changed interhemispheric relationships in cortical structure in some amputees.

Limb amputation and related changes in body feelings are associated with cortical functional reorganization that is reflected by increased interhemispheric asymmetry of body maps in the postcentral somatosensory cortex (PCS). As a pilot test to determine if limb amputation affects interhemispheric symmetry in PCS structure, we used MRI and computational morphometry to examine interhemispheric relationships of PCS thicknesses in a case series of eight lower limb amputees compared with 11 control subjects. As a further control, the same relationships were compared in the lateral occipital visual cortex (LOV) which, by nature of its visual connectivity, would be expected to be less related to amputation. The PCS thicknesses in the left and right hemispheres were positively related in control subjects, but not in amputees. The range of the PCS interhemispheric thickness differences (ID) in amputees was larger than the range in control subjects, and four of eight amputees had PCS ID that were at or above the maximal control subject ID. In contrast, LOV thicknesses in the two hemispheres were positively related and LOV ID ranges were similar in both amputees and control subjects. The results from this case series suggest the hypothesis that amputation alters PCS interhemispheric thickness relationships in some amputees. Further tests of this hypothesis would be useful to determine whether changes in structural symmetry contribute to known post-amputation alterations in PCS functional map symmetry and body feeling.

Ultra-sharp metal and nanotube-based probes for applications in scanning microscopy and neural recording.

This paper discusses several methods for manufacturing ultra-sharp probes, with applications geared toward, but not limited to, scanning microscopy (STM, AFM) and intra-cellular recordings of neural signals. We present recipes for making tungsten, platinum/iridium alloy, and nanotube fibril tips. Electrical isolation methods using Parylene-C or PMMA are described.

Neonatally elevated serotonin levels alter terminal arbors of individual retinal ganglion cells in superior colliculus of hamsters.

Previous studies from this laboratory showed that sprouting of serotoninergic (5-HT) axons in the hamster's superior colliculus (SC), induced by a single subcutaneous injection of 5,7-dihydroxytryptamine (5,7-DHT) at birth (postnatal day 0 [P-0]), resulted in an increased terminal distribution of the uncrossed retinocollicular projection that was not associated with any changes in the number or distribution of ipsilaterally projecting retinal ganglion cells. The present study was undertaken to determine what effect this manipulation had on the terminal arbors of such axons. Retinocollicular axons of normal and 5,7-DHT-treated animals were anterogradely labeled with small intraretinal injections of the lipophilic dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) on P-16. After tissue processing on P-19, single retinocollicular axon arbors were reconstructed by using confocal microscopy. Quantitative analysis indicated that arbors from 5,7-DHT-treated hamsters had significantly greater total fiber lengths, areas, and volumes than those from normal animals. There were no differences between axons from the two groups in number of branch points, distribution of relative branch lengths, and numbers of bouton-like swellings. These results support the hypothesis that increased SC concentrations of 5-HT alter development of the uncrossed retinocollicular pathway such that a greater territory is covered by individual terminal arbors but that the number of synaptic contacts per arbor remains constant. This may explain, at least in part, the abnormally widespread distribution of the aggregate ipsilateral projection.

Clorgyline treatment elevates cortical serotonin and temporarily disrupts the vibrissae-related pattern in rat somatosensory cortex.

Manipulation of cortical serotonin (5-HT) levels in perinatal rodents produces significant alterations in the development of the layer IV cortical representation of the mystacial vibrissae. Monoamine oxidase A (MAO(A)) knockout mice have highly elevated cortical 5-HT and completely lack barrels in somatosensory cortex (S-I). The present study was undertaken to determine whether the effects on thalamocortical development seen in MAO(A) knockout mice can be replicated in perinatal rats treated with an MAO(A) inhibitor and, second, to determine whether these effects persist with continued treatment or after discontinuation of the drug. Littermates were injected with either clorgyline (5 mg/kg) or sterile saline five times daily. Clorgyline administration from birth to postnatal day (P) 6, 8, or 10 produced increases of 1,589.4 +/- 53.3%, 1660.2 +/- 43.1% and 1,700.5 +/- 84.5 %, respectively, in cortical 5-HT as compared with controls. Serotonin immunocytochemistry, 1,1;-dioctadecyl-3,3,3", 3;-tetramethylindocarbocyanine perchlorate (DiI) labeling of thalamocortical afferents and Nissl and cytochrome oxidase staining of layer IV cellular aggregates demonstrated that clorgyline treatment from P0 to P6 produced a complete absence of any segmentation of vibrissae-related patches in S-I. However, continued treatment until P8 or P10 did not prevent the appearance of these patches. Animals treated with clorgyline from birth to P6 and killed on P8 or P10 had increases of 546.8 +/- 33.2% and 268.8 +/- 6.3% in cortical 5-HT and they had qualitatively normal vibrissae-related patterns in S-I. These results indicate that clorgyline treatment produces a transient disruption of vibrissae-related patterns, despite the continued presence of elevated cortical 5-HT.

The cortical vibrissae representation is normal in transgenic mice lacking the 5-HT(1B) receptor.

Recent studies have suggested that 5-HT may modulate thalamocortical development in somatosensory cortex (S-I) of rats and mice, and that the 5-HT(1B) receptor may play a critical role in this process. Analysis of CO-stained sections through lamina IV of S-I in perinatal and adult 5-HT(1B) knockout mice revealed a normal vibrissae-related pattern, indicating that activation of the 5-HT(1B) receptor is not necessary for the normal development of the vibrissae representation in S-I.

Effects of norepinephrine on the activity of visual neurons in the superior colliculus of the hamster.

Single-unit recording and micropressure ejection techniques were used to test the effects of norepinephrine (NE) on the responses of neurons in the superficial layers (the stratum griseum superficiale and stratum opticum) of the hamster's superior colliculus (SC). Application of NE suppressed visually evoked responses by -30% in 75% of 40 neurons tested and produced > or = 30% augmentation of responses in only 5%. The decrement in response strength was mimicked by application of the alpha2 adrenoceptor agonist, p-aminoclonidine, the nonspecific beta agonist, isoproterenol, and the beta1 agonist, dobutamine. These agents had similar effects on responses evoked by electrical stimulation of the optic chiasm and visual cortex. The alpha1 agonist, methoxamine, augmented the light-evoked responses of 53% of 49 SC cells by > or = 30%, but had little effect on responses evoked by electrical stimulation of optic chiasm or visual cortex. The effects of adrenergic agonists upon the glutamate-evoked responses of SC cells that were synaptically "isolated" by concurrent application of Mg2+ were similar to those obtained during visual stimulation. Analysis of effects of NE on visually evoked and background activity indicated that application of this amine did not significantly enhance signal-to-noise ratios for most superficial layer SC neurons, and signal-to-noise ratios were in some cases reduced. These results indicate that NE acts primarily through alpha2 and beta1 receptors to suppress the visual responses of SC neurons. Activation of either of these receptors reduces the responses of SC neurons to either of their two major visual inputs as well as to direct stimulation by glutamate, and it would thus appear that these effects are primarily postsynaptic.

Effects of norepinephrine upon superficial layer neurons in the superior colliculus of the hamster: in vitro studies.

Intracellular recording techniques were used to evaluate the effects of norepinephrine (NE) on the membrane properties of superficial layer (stratum griseum superficiale and stratum opticum) superior colliculus (SC) cells. Of the 207 cells tested, 44.4% (N = 92) were hyperpolarized by > or = 3 mV and 8.7% (N = 18) were depolarized by > or = 3 mV by application of NE. Hyperpolarization induced by NE was dose dependent (EC50 = 8.1 microM) and was associated with decreased input resistance and outward current which had a reversal potential of -94.0 mV. Depolarization was associated with a very slight rise in input resistance and had a reversal potential of -93.1 mV for the single cell tested. Pharmacologic experiments demonstrated that isoproterenol, dobutamine, and p-aminoclonidine all hyperpolarized SC cells. These results are consistent with the conclusion that NE-induced hyperpolarization of SC cells is mediated by both alpha2 and beta1 adrenoceptors. The alpha1 adrenoceptor agonists, methoxamine and phenylephrine, depolarized 35% (6 of 17) of the SC cells tested by > or = 3 mV. Most of the SC cells tested exhibited responses indicative of expression of more than one adrenoceptor. Application of p-aminoclonidine or dobutamine inhibited transsynaptic responses in SC cells evoked by electrical stimulation of optic tract axons. Inhibition of evoked responses by these agents was usually, but not invariably, associated with a hyperpolarization of the cell membrane and a reduction in depolarizing potentials evoked by application of glutamate. The present in vitro results are consistent with those of the companion in vivo study which suggested that NE-induced response suppression in superficial layer SC neurons was primarily postsynaptic and chiefly mediated by both beta2 and beta1 adrenoceptors.

Increased serotonin in the developing superior colliculus affects receptive-field size of retinotectal afferents but not that of postsynaptic neurons.

Administration of a single subcutaneous dose of 5,7-dihydroxytryptamine (5,7-DHT) to newborn hamsters results in a significant increase in the density of serotoninergic (5-HT) fibers in the superficial layers of the superior colliculus (SC) and marked abnormalities in the uncrossed retinotectal projection when these animals reach adulthood (Rhoades et al., 1993). The present study was undertaken to determine whether elevation of 5-HT in the developing SC altered the visual representation in SC. Multi-unit recordings from SC cells demonstrated that the overall organization of the visual map in the superficial SC laminae was normal and that the receptive-field sizes for unit clusters were unchanged in the 5,7-DHT-treated animals. However, when a combination of CNQX and MK-801 was directly applied to the SC to block postsynaptic activity, the receptive fields of unit clusters (presumably retinotectal axon terminals) in the 5,7-DHT treated animals were significantly larger than those in the normally reared hamsters. These results are consistent with the conclusions that elevation of 5-HT levels in the developing SC reduces the postnatal refinement of the crossed retinotectal axons, and that mechanisms operating within the SC may act to maintain normal sizes for the receptive fields of its constituent neurons.

Organization of the corticotectal projection in hamsters with neonatally elevated levels of serotonin in the superior colliculus.

A previous study from this laboratory showed that elevated serotonin (5-HT) levels in the hamsters superior colliculus (SC), induced by a single subcutaneous injection of 5,7-dihydroxytryptamine (5,7-DHT) at birth, resulted in an abnormally widespread distribution of the uncrossed retinotectal projection. The present study investigated whether the corticotectal projection in such animals was altered. Adult normal and 5,7-DHT-treated hamsters were injected with horseradish peroxidase (HRP) into occipital cortex and processed for anterograde tracing of corticotectal terminals in the SC. Quantitative analysis showed that normal and 5,7-DHT-treated hamsters were not significantly different in total labeling or in the gradient of labeling density within the SC. These data indicate that corticotectal axons achieve normal terminal fields after neonatal elevation of 5-HT in the SC.

Augmentation of serotonin in the developing superior colliculus alters the normal development of the uncrossed retinotectal projection.

A previous study from this laboratory showed that sprouting of serotoninergic axons in the hamster's superior colliculus (SC) induced by a single subcutaneous injection of 5,7-dihydroxytryptamine (5,7-DHT) at birth (postnatal day 0; P-0) resulted in an abnormal terminal distribution of the uncrossed retinotectal projection. The present study provided further evidence to support the role of increased 5-HT levels within the SC in this phenomenon. Slow-release polymer (ELVAX) chips impregnated with serotonin (5-HT) were placed over the SC on either P-1 or P-3, and retinotectal projections were assessed via anterograde transport of horseradish peroxidase when animals reached P > 18. Analysis of ELVAX chips indicated that they released 5-HT in amounts of > or = 1 pmole/hour for at least 12 days. Assessment of the SC of treated hamsters indicated significantly elevated 5-HT concentrations as late as P-12, but not on P-16. Implantation of 5-HT chips, but not control chips, resulted in abnormalities in the uncrossed retinotectal projection similar to those observed in the 5,7-DHT-treated animals. The patches that normally develop in the rostral part of the stratum opticum were not present, and uncrossed axons were distributed densely in this layer and in the lower portion of the stratum griseum superficiale throughout the rostrocaudal and mediolateral extents of the SC. Quantitative analysis of these changes indicated significant differences between the organization of the uncrossed retinotectal projections of 5-HT-treated animals vs. either blank-implant treated or completely untreated animals but not between 5-HT-treated hamsters and animals that received neonatal 5,7-DHT injections. All of these results support the conclusion that increased SC concentrations of 5-HT altered retinotectal development.

Effects of neurotensin on visual neurons in the superficial laminae of the hamster's superior colliculus.

Autoradiography with 125I-neurotensin in normal and enucleated hamsters was used to define the distribution of receptors for this peptide in the superficial layers of the superior colliculus (SC). Neurotensin binding sites were densely distributed in the stratum griseum superficiale (SGS), and results from the enucleated animals indicated that they were not located on retinal axons. The effects of neurotensin on individual superficial layer cells were tested in single-unit recording experiments. Neurotensin was delivered via micropressure ejection during visual stimulation (n = 75 cells), or during electrical stimulation of either the optic chiasm (OX; n = 47 cells) or visual cortex (CTX; n = 29 cells). In comparison with control values, application of neurotensin decreased visual responses of all SC cells tested to 54.1 +/- 34.9% (mean +/- standard deviation; range of decrement 7.5 to 100%; nine cells showed no effect or an increase in visual activity, which for four of these was > or = 30%). Neurotensin application also reduced responses to electrical stimulation of either OX or CTX, respectively, to 65.8 +/- 36.5% of control values (range of decrement 2.6 to 97.4%; 12 neurons showed a weak increment < or = 30%) and 68.0 +/- 38.5% (range of decrement 3.3 to 100%; five cells showed no effect or an increment, in one case > or = 30%). Of the 25 neurons tested with both OX and CTX stimulation, the correlation of evoked response suppression by neurotensin was highly significant (r = 0.70; P < 0.001). This suggests that the suppressive effects of neurotensin were common to both pathways. To test whether the inhibitory effects of neurotensin were presynaptic or postsynaptic, Mg2+ ions were ejected iontophoretically to abolish synaptic responses, and the neurons (n = 16) were activated by iontophoresis of glutamate and then tested with neurotensin. Neurotensin reduced the glutamate-evoked responses to an average 59.3 +/- 37.9% of control values (range 2.3 to 92.5%; one cell showed an increment > 30%). This result suggests that the site of action of neurotensin is most likely postsynaptic.

Increased serotonin in the developing superior colliculus does not alter the number or distribution of retinotectal ganglion cells.

Administration of a single subcutaneous dose of 5,7-dihydroxytryptamine (5,7-DHT) to newborn hamsters results in a significant increase in the density of serotoninergic (5-HT) fibers in the superficial layers of the superior colliculus (SC) and marked abnormalities in both the crossed and uncrossed retinotectal projections when these animals reach adulthood (R. Rhoades, C. Bennett-Clarke, R. Lane, M. Leslie, and R. Mooney, 1993, J. Comp. Neurol. 334:397-409). The present study was undertaken to determine whether changes in the retinotectal projection of 5,7-DHT-treated animals were associated with alterations in the number or distribution of retinal ganglion cells in these animals. Nissl staining of retinae from normal adult and 5,7-DHT-treated hamsters revealed no differences between them in the number or average diameter of cells in the retinal ganglion cell layer. Retrograde labeling with horseradish peroxidase (HRP) demonstrated no effect of 5,7-DHT treatment on the number or distribution of ipsilaterally or contralaterally projecting ganglion cells. Neonatal 5,7-DHT administration also had no effect on the distribution of soma diameters for HRP-labeled retinal ganglion cells. Electron microscopic analysis demonstrated no significant difference between the number of optic nerve fibers in the normal and 5,7-DHT-treated hamsters. The results are consistent with the conclusion that the effect of 5,7-DHT on the retinotectal projection may primarily be a function of this toxin, or the increase in 5-HT it induces, on the terminal arbors of retinotectal axons rather than on their parent cells.

Serotonin modulates retinotectal and corticotectal convergence in the superior colliculus.

A dense serotonin (5-HT)-containing projection to the superficial layers of the superior colliculus (SC) has been demonstrated in diverse mammalian species, but how 5-HT may affect visual signals within these laminae is largely unknown. This study undertook to investigate the distribution of 2 types of 5-HT receptors in the SC and to ascertain their physiological effects on transmission of visual signals to the SC from the retinotectal and corticotectual pathways. Autoradiography of tissue sections exposed to [3H]-8-OH-DPAT (8-hydroxy-dipropylaminotetraline) or to [125I]cyanopindolol plus isoproterenol showed that 5-HT1A and 5-HT1B receptors, respectively, were present in the superficial SC layers. In unilaterally enucleated animals, binding of ligand to 5-HT1B receptors was greatly reduced on the deafferented (contralateral) side, which is consistent with the possibility that these receptors are located on preterminal axons. Binding to 5-HT1A receptors was unaltered by enucleation. In recordings of superficial layer neurons from SC slices, application of 5-HT during blockade of 5-HT1A receptors with spiperone reduced the amplitude of EPSPs evoked by stimulation of the optic tract. The 5-HT concentration for a 50% reduction in EPSP amplitude was 6 microM. Under these conditions, there were no significant alterations in either membrane potential or input resistance concurrent with 5-HT mediated reduction in EPSPs. During extracellular in vivo recordings, 5-HT, applied by iontophoresis or micropressure or by endogenous release produced by electrical stimulation of the dorsal raphé nucleus, strongly suppressed visual activity in SC neurons. The effectiveness of 5-HT application was significantly stronger on responses evoked by electrical stimulation of the optic chiasm (an average response decrement of 92.2%) than on these evoked in the same neurons by stimulation of visual cortex (an average response reduction of 32.3%). These results support the following conclusions. The 5-HT1B receptors are located preferentially on optic axon terminals and exert presynaptic inhibition of retinotectal inputs. Secondly, 5-HT1A receptors probably have a postsynaptic localization and may affect activity of SC neurons irrespective of the source of input. The combined effect of 5-HT at both subtypes would bias SC visual activity toward information received from the corticotectal pathway.

Effects of neonatal and adult enucleation on the synaptic organization of the serotoninergic projection to the superficial gray layer of the hamster's superior colliculus.

Previous experiments from this laboratory have demonstrated that either neonatal or adult enucleation results in a marked increase in the density of the serotoninergic (5-HT) projection to the superficial layers of the hamster's superior colliculus (SC). The present study was undertaken to determine whether this alteration in the density of the 5-HT projections to SC in hamsters that sustained eye removals was associated with any change in the synaptic organization of this pathway. In normal adult hamsters, only 4.0% of 500 5-HT-immunoreactive profiles made synaptic contacts in the stratum griseum superficiale (SGS). Of these, 65% were axoaxonic, 30% were axodendritic, and 5% were axosomatic. In contrast, 22% of 400 5-HT-positive profiles in the neonatally enucleated hamsters and 17.5% of 400 such profiles in hamsters that sustained eye removals in adulthood made synaptic contacts. In the neonatal enucleates, 66% of the contacts were axoaxonic, 33% were axodendritic, and 1% were axosomatic. In the adult enucleates, 73% of the contacts were axoaxonic, 26% were axodendritic, and 1% were axosomatic. Thus, both neonatal and adult enucleation increase the percentage of 5-HT-positive profiles that make synaptic contacts in SC, but neither manipulation significantly changes the distribution of these contacts among different types of postsynaptic targets.

Neonatal administration of the neurotoxin 5,7-dihydroxytryptamine results in synaptic reorganization in the superficial gray layer of the hamster's superior colliculus.

Neonatal subcutaneous administration of the neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) to hamsters results in a marked depletion of serotonin (5-HT) in cortex and an increase in the concentration of this amine in the superior colliculus (SC). To determine whether this increase was associated with an alteration in the synaptic organization of 5-HT-containing axons in the superficial gray layer of the SC, immunocytochemistry was combined with electron microscopy. In normal adult hamsters, only 4.0% of 500 5-HT-immunoreactive profiles make synaptic contacts in the superficial gray layer of the hamster's SC. In 5,7-DHT-treated animals, examination of 400 individual profiles indicated that 25.5% of 5-HT-positive profiles made synaptic contacts (P < 0.05). Given the recently demonstrated effect of 5-HT on retinotectal transmission in this species, the present results suggest that the functional organization of the SC may also be markedly altered in animals that sustain neonatal 5,7-DHT administration.

Effects of angiotensin II on visual neurons in the superficial laminae of the hamster's superior colliculus.

Superficial layer superior colliculus (SC) neurons were recorded extracellularly with multibarreled recording/ejecting micropipettes. Angiotensin II was delivered via micropressure ejection during visual stimulation (n = 215 cells), or during electrical stimulation of either the optic chiasm (OX; n = 150 cells) or visual cortex (CTX; n = 42 cells). Application of angiotensin II decreased visual responses of SC cells to 43.8% +/- 30.7% (mean +/- S.D.) and reduced responses to electrical stimulation of the OX and CTX to 58.6% +/- 34.1% and 43.8% +/- 30.7% of control values, respectively. Angiotensin II enhanced responses by at least 30% in only 6 cells (1.5%). Of the 35 neurons tested with both OX and CTX stimulation, the correlation of evoked response suppression by angiotensin II was highly significant (r = 0.69; P < 0.001). This suggests that the suppressive effects of angiotensin II were common to both pathways. To test whether the inhibitory effects of angiotensin II were presynaptic or postsynaptic, Mg2+ ions were ejected iontophoretically to abolish synaptic responses, and the neurons were activated by iontophoresis of glutamate and then tested with angiotensin II. Angiotensin II reduced the glutamate-evoked responses to an average 29.1% +/- 21.1% of control values (n = 9 cells). This suggest that the site of action of angiotensin II is most likely postsynaptic. To identify which receptors were involved in these effects, angiotensin II was ejected concurrently with the AT1 antagonist Losartan (DUP753) or with either of two AT2 antagonists, CGP42112A or PD123177. Losartan antagonized the action of angiotensin II in 65.6% of the cells tested (n = 99) and CGP42112A and PD123177 had antagonistic effects in 58% (n = 65) and 60% (n = 5), respectively. Both classes of antagonists were tested in 29 cells; and there was no significant correlation between their effectiveness. These results suggest that both AT1 and AT2 receptors may independently mediate the suppressive effects of angiotensin II, and that collicular neurons may have either or both receptor subtypes.

Effects of 5-HT on thalamocortical synaptic transmission in the developing rat.

1. Recent immunocytochemical and receptor binding data have demonstrated a transient somatotopic patterning of serotonin (5-HT)-immunoreactive fibers in the primary somatosensory cortex of developing rats and a transient expression of 5-HT1B receptors on thalamocortical axons from the ventral posteromedial thalamic nucleus (VPM). 2. These results suggest that 5-HT should strongly modulate thalamocortical synaptic transmission for a limited time during postnatal development. This hypothesis was tested in intracellular recording experiments carried out in thalamocortical slice preparations that included VPM, the thalamic radiations, and the primary somatosensory cortex. Effects of 5-HT and analogues were monitored on membrane potentials and input resistances of cortical neurons and on the amplitude of the synaptic potentials evoked in them by stimulation of VPM. 3. Results obtained from cortical neurons in slices taken from rats during the first 2 wk of life indicated that 5-HT strongly inhibited the VPM-evoked excitatory postsynaptic potential (EPSP) recorded from cortical neurons in a dose-dependent manner. In contrast, 5-HT had no significant effects on membrane potential, input resistance, or depolarizations induced by direct application of glutamic acid to cortical cells. 4. The effects of 5-HT were mimicked by the 5-HT1B receptor agonists 1-[3-(trifluoromethyl)phenyl]-piperazine (TFMPP) and 7-trifluoromethyl-4(4-methyl-1-piperazinyl)-pyrrolo[1,2-a]-quinoxaline maleate and antagonized by the 5-HT1B receptor antagonist (-)-pindolol. The 5-HT1A agonist [(+/-)8-hydroxydipropylaminotetralin HBr] (8-OH-DPAT) had less effect on the VPM-elicited EPSP, and the effects of 5-HT upon this response were generally not antagonized by either 1-(2-methoxyphenyl)-4-[4-(2- phthalimmido)butyl]piperazine HBr (a 5-HT1A antagonist) or ketanserine (a 5-HT2 antagonist) or spiperone (a 5-HT1A and 2 antagonist). 5. The ability of 5-HT to inhibit the VPM-evoked EPSP in cortical neurons was significantly reduced in slices from animals > 2 wk of age. The effectiveness of TFMPP in such animals was even more attenuated than that of 5-HT, and the effectiveness of 8-OH-DPAT was unchanged with age. These results are consistent with the disappearance of 5-HT1B receptors from thalamocortical axons after the second postnatal week and the maintenance of 5-HT1A receptors on some neurons. 6. All of the results obtained in this study are consistent with the conclusion that 5-HT has a profound, but developmentally transient, presynaptic inhibitory effect upon thalamocortical transmission in the rat's somatosensory cortex.

Modulation of retinotectal transmission by presynaptic 5-HT1B receptors in the superior colliculus of the adult hamster.

1. Radioligand binding with [125I]-cyanopindolol in the presence of isoproterenol was used to define the distribution of 5-HT1B receptors in the superior colliculus (SC) of adult hamsters. There was a high density of these receptors in the stratum griseum superficiale (SGS), and they were much less dense in other SC laminae. Enucleation of one eye produced a marked reduction in the density of these receptors in the contralateral SGS, suggesting that they are located primarily on retinotectal axon terminals. 2. Intracellular recording techniques were used to evaluate the effects of serotonin (5-HT) on the excitatory postsynaptic potentials (EPSPs) evoked in SC cells of adult hamsters by stimulation of the optic tract (OT) in vitro. Application of 5-HT produced a reduction of > or = 50% in OT-evoked EPSPs in 79% of the 67 cells tested. The average EPSP amplitude was 7.8 +/- 2.1 (SD) mV under control conditions and 2.7 +/- 1.9 mV in the presence of 5-HT (P < 0.01). For most of these neurons, application of 5-HT had little effect on their membrane potential or input resistance. The average percent change in membrane potential for cells tested with 5-HT was 0.5 +/- 6.0% and the average percent change in input resistance was 0.6 +/- 22.9%. 3. For four of six cells tested, application of 5-HT had no significant effects on the responses evoked by application of glutamate, either under normal bathing conditions or when the medium included low Ca2+ and high Mg2+. 4. Pharmacologic experiments indicated that the effects of 5-HT on retinotectal transmission were mimicked by the 5-HT1B agonists 1-[3-(trifluoromethyl)phenyl]-piperazine and 7-trifluoromethyl-4(4-methyl-1-piperazinyl) [1,2-a]-quinoxaline maleate and antagonized by the 5-HT1A/1B antagonists (-)-pindolol and methiothepin. The effects of 5-HT on the OT-evoked EPSP were not antagonized by either spiperone, ketanserin, 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl]-piperazine HBr, or [1-H-3 alpha-5 alpha-tropan-3-yl]-3,5-dichlorobenzoate. 5. Both the anatomic and physiological results are consistent with the conclusion that 5-HT presynaptically inhibits retinotectal transmission and that this effect is mediated by the 5-HT1B receptor.

Correlations between the receptive field properties and morphology of neurons in the deep layers of the hamster's superior colliculus.

Extracellular and intracellular recording, receptive field mapping, and intracellular HRP injection techniques were used to define the morphological classes of cells in the deep laminae of the hamster's superior colliculus and to determine whether there are any correlations between the structural and functional characteristics of these neurons. A total of 110 neurons were characterized and reconstructed. Of these, 23.6% (N = 26) were visual, 60% (N = 66) were somatosensory, 0.9% (N = 1) were bimodal (visual-somatosensory), and 15.4% (N = 17) were unresponsive. Of the somatosensory neurons, 72.7% (N = 48) were low threshold, 4.5% (N = 3) had a wide dynamic range, 9.1% (N = 6) responded only to noxious stimulation, and 13.6% (N = 9) had complex somatosensory receptive fields. Deep layer cells were divided into eight morphological classes. These classes were multipolar cells (26.4%, N = 29), bipolar cells (9.1%, N = 10), widefield vertical cells (7.3%, N = 8), horizontal cells (13.6%, N = 15), stellate cells (10.9%, N = 12), ventrally directed cells (5.5%, N = 6), sparse radial cells (17.3%, N = 19), and small sparse radial cells (6.4%, N = 7). Four cells (3.6%) did not fit into this classification scheme. Univariate and multivariate analyses of variance of properties such as soma area, number of branch points, total dendritic length, and volume and orientation of dendritic arbor indicated that these classes were significantly different. However, chi 2 analysis and multivariate analysis of variance indicated no significant relationships between morphological class and either laminar location or receptive field type. There was a significant positive relationship between the possession of dendrites that extended into the superficial laminae and visual responsivity.

Reduction in the percentage of serotoninergic axons making synapses during the development of the superficial layers of the hamster's superior colliculus.

Immunocytochemistry with an antibody directed against a serotonin (5-HT)-bovine serum albumin conjugate was combined with electron microscopy to assess the synaptic organization of the serotoninergic projection to the stratum griseum superficiale (SGS) of the superior colliculus of hamsters killed on postnatal days (P) 0, 3, 7, 12, 15, 20 and > 60. At least 200 5-HT-immunoreactive profiles were examined at each of these ages. In the newborn (P0) animals, 36.6% of the 5-HT-positive profiles made conventional synapses. This percentage remained fairly constant until P15 when only 17.3% of the profiles made conventional synaptic contacts. On P20, this value decreased to 10.5% and in the adult animals, it fell to 4.4%. These results thus indicate a dramatic age-related change in the synaptic organization of the 5-HT input to the hamster's superior colliculus.