Short-term lithium treatment promotes neuronal survival and proliferation in rat striatum infused with quinolinic acid, an excitotoxic model of Huntington's disease.

We assessed the ability of lithium to reduce neurodegeneration and to stimulate cell proliferation in a rat model of Huntington's disease in which quinolinic acid (QA) was unilaterally infused into the striatum. LiCl (0.5-3.0 mEq/kg) was injected subcutaneously 24 h before and 1 h after QA infusion. At 7 days after QA injection, lithium significantly diminished the loss of neurons immunostained for Neuronal Nuclei (NeuN) in the injured striatum, but failed to prevent the reduction of NADPH-diaphorase-positive striatal interneurons. Lithium also reduced the number of neurons showing DNA damage or activated caspase-3. This neuroprotection was associated with an upregulation of Bcl-2 protein levels in the striatal tissue and an increase in the number and density of Bcl-2 immunostaining in striatal neurons. Bromodeoxyuridinie (BrdU) labeling in the lithium-treated injured striatum revealed the presence of large numbers of proliferating cells near the QA-injection site, with a reduction of BrdU-labeled cells in the subventricular zone (SVZ). All BrdU-labeled cells in the SVZ and the majority of BrdU-labeled cells near the QA-injection site were negative for either NeuN or glial fibrillary acidic protein (GFAP), suggesting that they are undifferentiated progenitor cells. However, a small number of BrdU-positive cells found in the QA-injected and lithium-treated striatum site were positive for either NeuN or GFAP. Our results suggest that lithium is neuroprotective in the QA-injection model of Huntington's disease not only due to its ability to inhibit apoptosis but also because it can stimulate neuronal and astroglial progenitor proliferation in the QA-injected striatum or their migration from the SVZ.

Distinct forms of cholinergic modulation in parallel thalamic sensory pathways.

Mammalian thalamus is a critical site where early perception of sensorimotor signals is dynamically regulated by acetylcholine in a behavioral state-dependent manner. In this study, we examined how synaptic transmission is modulated by acetylcholine in auditory thalamus where sensory relay neurons form parallel lemniscal and nonlemniscal pathways. The former mediates tonotopic relay of acoustic signals, whereas the latter is involved in detecting and transmitting auditory cues of behavioral relevance. We report here that activation of cholinergic muscarinic receptors had opposite membrane effects on these parallel synaptic pathways. In lemniscal neurons, muscarine induced a sustained membrane depolarization and tonic firing by closing a linear K(+) conductance. In contrast, in nonlemniscal neurons, muscarine evoked a membrane hyperpolarization by opening a voltage-independent K(+) conductance. Depending on the level of membrane hyperpolarization and the strength of local synaptic input, nonlemniscal neurons were either suppressed or selectively engaged in detecting and transmitting synchronized synaptic input by firing a high-frequency spike burst. Immunohistochemical and Western blotting experiments showed that nonlemniscal neurons predominantly expressed M2 muscarinic receptors, whereas lemniscal cells had a significantly higher level of M1 receptors. Our data indicate that cholinergic modulation in the thalamus is pathway-specific. Enhanced cholinergic tone during behavioral arousal or attention may render synaptic transmission in nonlemniscal thalamus highly sensitive to the context of local synaptic activities.

Extracortical descending projections to the rat inferior colliculus.

Feedback controlling is an important element in the sensory processing in the auditory system. It has been long recognized that the inferior colliculus (IC) sends direct ascending projections to the medial geniculate body (MGB), but receives feedback regulation from the auditory cortex. In the present study we probed the shorter extracortical projections to the IC, including the direct descending pathway from the MGB. In the rat, the fluorescence retrograde tracers Fluorogold, True Blue or Rhodamine latex microspheres were injected into the IC, and the auditory thalamus and surrounding regions were examined for fluorescent neurones. We did not find any retrograde labelling in the ventral division of the MGB. However, retrogradely labelled neurones were found in the medial and suprageniculate nuclei of the MGB. We also observed densely packed groups of fluorescent neurones in the peripeduncular nucleus and numerous labelled neurones in the nucleus of the brachium of the IC. The existence of a direct descending pathway to the IC from at least some auditory thalamic nuclei challenges the perception of the colliculo-thalamic relationship as one-way traffic and suggests more direct involvement of the auditory thalamus in the feedback regulation of the incoming acoustic signals.

Lithium suppresses excitotoxicity-induced striatal lesions in a rat model of Huntington's disease.

Huntington's disease is a progressive, inherited neurodegenerative disorder characterized by the loss of subsets of neurons primarily in the striatum. In this study, we assessed the neuroprotective effect of lithium against striatal lesion formation in a rat model of Huntington's disease in which quinolinic acid was unilaterally infused into the striatum. For this purpose, we used a dopamine receptor autoradiography and glutamic acid decarboxylase mRNA in situ hybridization analysis, methods previously shown to be adequate for quantitative analysis of the excitotoxin-induced striatal lesion size. Here we demonstrated that subcutaneous injections of LiCl for 16 days prior to quinolinic acid infusion considerably reduced the size of quinolinic acid-induced striatal lesion. Furthermore, these lithium pre-treatments also decreased the number of striatal neurons labeled with the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay. Immunohistochemistry and western blotting demonstrated that lithium-elicited neuroprotection was associated with an increase in Bcl-2 protein levels. Our results raise the possibility that lithium may be considered as a neuroprotective agent in treatment of neurodegenerative diseases such as Huntington's disease.

Sodium-potassium adenosine triphosphatase inhibition enhances membrane accumulation of DiI in rat hippocampus in vivo.

Transient brain ischemia induces significant alterations in lipid structures of neuronal membranes, which are believed to result from lipid peroxidation and free radical attack. Such a membrane structural change may serve as an important histological marker of cell injury. In the present study, we examined how the dynamics of DiI/membrane incorporation may reflect early membrane metabolism and dynamic changes following sodium-potassium pump inhibition. Ouabain (1mM) was stereotactically co-administered with either 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate DiI (50 microg/ml) or ethidium homodimer (4 microM) into the granule cell layer of the adult rat hippocampus. Tissue was cryosectioned and examined with epifluorescence microscopy at 1, 2, 3, 4, 6, 8 and 72h post-injection. Alternate sections were stained with thionine or haematoxylin and eosin to evaluate morphological changes. Ouabain-induced pump inhibition resulted in a dramatic increase in DiI fluorescence in granule cell layer neurons as early as 4h post-injection. This increase in DiI incorporation coincided both spatially and temporally with the appearance of reactive changes characterizing early neuronal injury. However, the fluorescence increase was not a result of membrane breakdown because ethidium homodimer, a membrane-impermeable nucleic acid probe used for labeling cells with compromised membranes, when applied in a similar fashion, failed to show any fluorescence changes. The results of this study suggest that pump inhibition results in a specific increase in membrane lipophilicity possibly due to altered lipid structure.

Regulation of Na+,K+-ATPase by persistent sodium accumulation in adult rat thalamic neurones.

The present study investigated the regulatory mechanism of the Na+, K+-ATPase and the level of internal Na+ and Ca2+ in response to persistent Na+ influx in acutely dissociated rat thalamic neurones. Whole-cell patch-clamp recordings and Na+ imaging revealed a stable [Na+]i and low background pump activity. Exposure to veratridine (50 microM) for 1 h resulted in a progressive rise in [Na+]i (DeltaFNa = 64 +/-22%) and [Ca2+]i (DeltaFCa = 44 +/- 14%) over 3 h. Increases in [Na+]i and [Ca2+]i were also observed during neuronal exposure to the Na+ ionophore monensin (50 microM). Subcellular confocal immunofluorescence quantification of alpha3 catalytic Na+-K+ pump subunits showed that a veratridine-induced rise in [Na+]i was accompanied by a significant increase in pump density in both membrane and cytoplasmic compartments, by 39 and 54%, respectively. Similar results were also obtained in experiments when neurones were treated with monensin. A fluorescent 9-anthroylouabain binding assay detected a 60 and 110% increase in phosphorylated (active) pumps after veratridine and monensin exposure, respectively. During the entire experiment, application of ouabain or veratridine alone induced little cell swelling and death, but pump inhibition in cells pre-loaded with Na+ led to rapid cell swelling and necrosis. The above results indicate that a persistent influx of Na+ may trigger rapid enhancement of pump synthesis, membrane redistribution and functional activity. However, these compensatory mechanisms failed to prevent persistent Na+ accumulation.

Ethidium bromide staining reveals rapid cell dispersion in the rat dentate gyrus following ouabain-induced injury.

The dentate gyrus of the hippocampal formation undergoes extensive network remodelling including progenitor cell proliferation, migration as well as apoptotic cell death in response to prolonged excitotoxic insults. Previous studies have shown that such a proliferative cell population may undergo aberrant migration and later persist in ectopically located positions within the molecular cell layer. In this study we have developed an experimental model to characterize the spatiotemporal patterns of such an injury-induced network remodelling. Ouabain (1 microl, 1 mM), a Na+, K+-ATPase blocker, was stereotactically co-injected into the rat dentate gyrus with ethidium bromide (1 microl, 40 microM). The latter is a fluorescent nucleic acid intercalating dye, which was used for labeling cells undergoing early phases of apoptosis or proliferation. Our results revealed that within an hour after the injection, a subpopulation of cells characterized by spindle- or ovoid-shaped somata and bipolar morphology, were intensely labeled with ethidium bromide. These cells were found initially clustered both inside and outside the dentate granule cell layer and later on markedly increased in number as well as spread radially in the next few hours into the dentate molecular layer. The unusual pattern of cell dispersion encountered in our study may represent aberrantly migrating progenitor cells consistent with earlier observations of ectopically-located granule cells in human temporal lobe epilepsy specimens and epilepsy animal models. Alternatively, the described phenomenon may represent dispersion of Cajal-Retzius cells that may be involved in post-lesion network remodelling.

Projections of medullary and pontine noradrenergic neurons to the horizontal limb of the nucleus of diagonal band in the rat.

Recent investigations in the rat have implicated a noradrenergic innervation to the horizontal nucleus of the diagonal band of Broca as a critical link in a neural circuit that conveys baroreceptor information centrally to inhibit the firing of vasopressin-secreting neurons in the hypothalamic supraoptic nucleus. In this study we used small intra-diagonal band injections of a retrograde tracer, rhodamine latex microspheres, in combination with tyrosine hydroxylase histochemistry to identify brainstem noradrenergic cells contributing to this innervation. In three cases where tracer injections were limited to the horizontal limb of the diagonal band, we observed 20-50 double-labelled neurons ipsilaterally in the dorsal part of the locus coeruleus (A6) and the caudal nucleus tractus solitarius (A2), and bilaterally in the caudal ventrolateral medulla (A1). Double-labelled neurons were also noted in the ventral tegmental area (dopaminergic A10 cell group). Although all major brainstem noradrenergic cell groups contribute fibers to the horizontal limb of the nucleus of diagonal band, data from physiological studies suggest that the noradrenergic A2 neurons in the nucleus tractus solitarius are the most likely pathway through which it receives this baroreceptor information.

Unidirectional axonal transport in in vitro adult rat brain explants.

The present study examined the properties of anterograde and retrograde transport in central axonal pathways maintained in vitro. The commonly-used tracers biocytin, dextran rhodamine B, FluoroGold, True Blue or rhodamine latex microspheres were injected into the medial geniculate body or the inferior colliculus of the adult rat brain explant. Injection of biocytin into the inferior colliculus consistently resulted in extensive anterograde labelling of axonal trunks and terminals in the ipsilateral medial geniculate body and in the contralateral inferior colliculus. Labelled axons were obtained 2-3 h after the injection at a site 3-4 mm away from the injection site and could be found up to 1.5 mm below the explant surface. Despite massive anterograde labelling with biocytin, all the tracers applied in the gray or white matter failed to show retrograde transport. These results suggest that axonal transport can occur in an anterograde-selective fashion in adult brain explants in vitro.

Expression of cholecystokinin messenger RNA in reciprocally-connected auditory thalamus and cortex in the rat.

Cholecystokinin exerts a potent antiepileptic action in mammalian auditory system and undergoes seizure-mediated up-regulation. The present study investigated cholecystokinin messenger RNA expression in the reciprocally-connected auditory thalamus and cortex in the rat. Immunofluorescence in situ hybridization was performed using a 24-base cholecystokinin-messenger RNA oligonucleotide probe. Corticothalamic projection neurons were identified by means of the retrograde fluorescent tracer rhodamine latex microspheres injected into the medial geniculate body. In our experiments, cholecystokinin messenger RNA transcripts were found in about 80% of neurons located within the reciprocally-connected regions of the medial geniculate body and the auditory cortices. These observations provide evidence of cholecystokinin production in the reciprocally-connected regions of the auditory thalamus and cortex, the structures which jointly create the thalamo-corticothalamic circuit which has been implicated in seizure genesis.

The electrogenic effects of Na(+)-K(+)-ATPase in rat auditory thalamus.

1. The electrogenic effects of the Na(+)-K(+)-ATPase in thalamic neurones were investigated by means of intracellular and whole-cell patch-clamp recording techniques in rat medial geniculate body (MGB) maintained in vitro. 2. In twenty-six out of thirty-one neurones recorded intracellularly, application of the Na(+)-K+ pump inhibitor strophanthidin induced two different types of membrane depolarization: a small, reversible depolarization with a peak amplitude of 4 +/- 2.6 mV or a prolonged depolarization of large amplitude (48.6 +/- 9.0 mV) with or without a decrease in apparent membrane resistance. Blockade of glutamate receptors with kynurenic acid or 6-cyano-7-nitroquinoxaline-2,3-dione and (+/-)-2-amino-5-phosphonopentanoic acid did not prevent either type of pump response, but the large depolarization was not seen when the medium contained the sodium channel blocker TTX. 3. Whole-cell patch-clamp recording showed that the small membrane depolarization is mediated by an inward membrane current (39.00 +/- 5.70 pA) that exhibited a weak voltage dependence. An inward current of similar amplitude was also induced in MGB cells when the pipette solution contained nominally zero Na+ or when K+ was temporarily omitted from the extracellular medium. The large membrane depolarization or the corresponding membrane current was not observed in whole-cell conditions. 6. Transient inhibition of the electrogenic Na(+)-K(+)-ATPase consistently led to a change in the mode of synaptic transmission in MGB cells, during which the synaptically evoked burst response was either blocked or converted into a single spike discharge. 7. Taken together, these data suggest that blockade of the electrogenic pump produces a dual membrane effect in mammalian thalamic neurones: a small electrogenic membrane depolarization and a large depolarization response that can be prevented by artificially maintaining the transmembrane ionic gradients. The electrogenic activity of the Na(+)-K(+)-ATPase may play an important role in setting the mode of synaptic transmission in sensory thalamus.

Differential Na(+)-K(+)-ATPase activity in rat lemniscal and non-lemniscal auditory thalami.

1. Using whole-cell recording and confocal immunofluorescent microscopy, we have investigated the differential electrogenic activity, subunit expression and subcellular distribution of the Na(+)-K(+)-ATPase in the lemniscal (ventral) and non-lemniscal (dorsal) pathways of the rat medial geniculate body (MGB) in vitro. 2. Bath application of Na(+)-K(+)-ATPase inhibitors strophanthidin or dihydro-ouabain produced a transient, dose-dependent inward current or membrane depolarization which were significantly larger in dorsal MGB neurones than in ventral cells (45.9 +/- 6.45 vs. 24.3 +/- 4.1 pA; P < 0.05). Electrophysiological and morphometric measurements showed that the dorsal MGB neurones had a significantly lower input conductance and a smaller somata than their ventral counterparts. The level of the resting membrane potential also differed by about 6 mV between the two cell populations, with the dorsal cells being more hyperpolarized (-74.2 +/- 0.6 vs. -67.7 +/- 1.3 mV; P < 0.001). 3. Incubation of enzymatically dissociated MGB neurones with fluorescent monoclonal antibodies against alpha 1-alpha 3 isoforms of Na(+)-K(+)-ATPase showed that both dorsal and ventral cells expressed primarily alpha 3 subunits. Confocal laser scanning revealed, however, that the mean pixel density of alpha 3 fluorescent antibodies in the plasma membrane domain, but not in the cytoplasmic compartment, was about 40% higher in dorsal neurones than in the ventral cells (29.7 +/- 4.7 vs. 16.9 +/- 2.3 grey shadow per pixel; P < 0.05). 4. The above results suggest that the electrogenic activity of the Na(+)-K(+)-ATPase is differentially regulated between lemniscal and non-lemniscal auditory thalami through a mechanism that probably involves differential pump densities in the cell membrane.

Immunofluorescence in situ hybridization (IFISH) in neurones retrogradely labelled with rhodamine latex microspheres.

The method of non-radioactive in situ hybridization was developed as an alternative to radioactive assay because of the difficulties and disadvantages of the safety measures required, extensive time required for autoradiography (especially with 3H-labelled probes) and limited cellular resolution obtained using 32P- and 35S-labelled probes. This method holds great potential for studying functional anatomy of specific neuronal pathways if it can be used in conjunction with conventional tract tracing techniques. In this article we describe a simple method by which immunofluorescence in situ hybridization (IFISH) was jointly used with rhodamine latex microspheres (RLM) to trace the origin of the thalamic cholecystokininergic input in rat. RLM is a widely used retrograde fluorescence tracer and seems ideal for IFISH because: (1) it lacks aversive effect on the hybridization and immunocytochemical reactions, (2) it is resistant to the rather harsh tissue treatment required for IFISH, and (3) both the RLM and mRNA hybrids give fluorescence signals; therefore, the extent of signal co-localization can be conveniently and more accurately verified under an epifluorescence microscope. Success of the IFISH-RLM combination is chiefly limited by the quantity and availability of mRNA signals in the tissue. In our case, we used a digoxigenin (DIG)-labelled oligonucleotide probe, which through immunological amplification significantly enhanced the sensitivity of mRNA detection.

Mature but not fetal or neonatal rat superior cervical ganglion transplants survive in the cortex of adult rats.

The transplantation of catecholaminergic tissues is a possible therapy for parkinsonism. Central nervous tissue is suitable for transplantation only in the immature stage, whereas peripheral nervous tissue can also be transplanted when mature. The present study compares the development of fetal (17-20 embryonic day, E17-20), neonatal (1-3 postnatal day, P1-3) and mature (5-6-week-old) rat superior cervical ganglia after transplantation into the cerebral cortex of adult rats. The mature transplants survived in greater proportion and preserved their structural characteristics, although a considerable proportion of the neurons died. The perinatal transplants only survived sporadically, decreased in size and the surviving remnants failed to display a structure comparable to the adult ganglion in situ. Thus, the use of adult donors is not only a possibility but a necessity when superior cervical ganglion (probably any ganglion) is transplanted. This principle is radically different from that seen in the case of central nervous tissues, and can be understood by the analysis of the time curves of cell proliferation and programmed cell death (apoptosis) observed during the perinatal development of sympathetic ganglia.

Muscarine induces an anomalous inhibition of synaptic transmission in rat auditory thalamic neurons in vitro.

Muscarinic receptor-mediated inhibition of central synaptic transmission was studied in a monosynaptic pathway connecting the inferior colliculus and the auditory thalamus in in vitro rat brain explants. Extra- and intracellular synaptic responses were recorded by sharp electrode and whole-cell patch clamp techniques in the ventral nucleus of the medial geniculate body after electrical stimulation of the brachium of the inferior colliculus. Stimulation of tectal afferents evoked either a high-frequency burst or a single-spike synaptic response in ventral geniculate neurons. Bath application of muscarinic receptor agonists abolished responses consisting of a high-frequency burst, but not responses consisting of a single spike. In the majority of single-spike cells muscarinic agonists often induced a synaptic facilitation. The burst blocking effect was mimicked by a moderate elevation of extracellular potassium. Intracellular recordings showed that the burst synaptic responses similar to that recorded extracellularly were induced by an excitatory postsynaptic potential. This synaptic potential, by first activating a low-threshold spike, was able to evoke a burst of sodium spike discharges. Muscarinic agonists caused a slow membrane depolarization that inactivated the low-threshold spike, leading to a blockade of the burst response. This mechanism is tentatively termed here as EPSP-LTS decoupling. Our results therefore support the hypothesis that part of the muscarinic receptor-mediated synaptic inhibition previously reported in anesthetized animal preparations in vivo represents a membrane depolarization rather than pre- or postsynaptic inhibition.

Expression of cholecystokinin mRNA in corticothalamic projecting neurons: a combined fluorescence in situ hybridization and retrograde tracing study in the ventrolateral thalamus of the rat.

Cholecystokinin (CCK), a well-known neuroactive peptide, has been observed in the axon endings within the thalamic reticular nucleus and the adjacent ventrolateral nucleus of the thalamus. The origin of this CCK innervation remains undefined. In this study, a fluorescence in situ hybridization (FISH) technique was used in conjunction with latex microsphere retrograde tracing to investigate whether cortical neurons may provide a source of CCK afferents to the ventrolateral thalamic nucleus. Rhodamine latex beads were injected into the ventrolateral thalamic nucleus of adult male rats to retrogradely label corticothalamic cells. After 7 days, tissues were processed for FISH using a 24-base oligonucleotide probe complementary to the 3' coding region of rat preprocholecystokinin mRNA. It was found that CCK transcripts are expressed in about 80% of identified corticothalamic projecting neurons. We therefore conclude that the descending cortical projections to the ventrolateral thalamus may provide an important source of CCK innervation to this region of the brain.

Low extracellular magnesium unmasks N-methyl-D-aspartate-mediated graft-host connections in rat neocortex slice preparation.

The main purpose of this study was to investigate the role of N-methyl-D-aspartate receptors in host-graft synaptic transmission in the neocortex. The effects of low extracellular magnesium, the glutamate agonist N-methyl-D-aspartate and N-methyl-D-aspartate antagonists on the synaptic activation of connections between embryonic neocortical graft tissue and the surrounding host tissue were studied in 17 perfused slices of rat neocortex. In standard artificial cerebrospinal fluid, stimulation of the host white matter evoked field potentials in four of 17 grafts. However, in Mg(2+)-free medium, the same stimulation evoked field potentials in an additional six grafts, with significant increases in the mean duration of the evoked responses in the 10 responsive grafts. In five of these slices stimulation of the graft also evoked field potentials in the host tissue, suggesting reciprocal interaction between graft and host. Simultaneous extracellular recordings from graft and host tissues in Mg(2+)-free medium showed that spontaneous epileptiform discharges developed in the graft and host tissue synchronously. In Mg(2+)-free medium, application of N-methyl-D-aspartate induced a shift of the baseline with superimposed epileptiform discharges in both graft and host. Application of the non-competitive N-methyl-D-aspartate antagonist ketamine and the competitive antagonist D,L-2-amino-5-phosphonovaleric acid attenuated or reversibly blocked both the spontaneous epileptiform discharges and the evoked field potentials. Our data provides evidence that N-methyl-D-aspartate receptors are present at synapses created between fetal graft and host neocortex, and that the N-methyl-D-aspartate-activated receptor-channel complex plays an active role in mediating excitatory synaptic transmission in host-graft circuitry.

Organum vasculosum lamina terminalis-evoked postsynaptic responses in rat supraoptic neurones in vitro.

1. To characterize the organum vasculosum lamina terminalis (OVLT) innervation of hypothalamic supraoptic nucleus (SON) neurones, current clamp recordings were obtained in SON cells in superfused rat hypothalamic explants. Stimulation of 1 Hz evoked 5-10 mV bicuculline-sensitive IPSPs in forty out of forty-six SON neurones, including both phasic (vasopressin immunoreactive) and continuously firing (oxytocin immunoreactive) cells. 2. In twenty-four cells, mean IPSP latency was 8.7 +/- 1 ms (+/- S.D.) and reversal potentials (Vr) ranged between -60 and -75 mV. In the other sixteen cells, Vr ranged between -20 and -55 mV and the addition of bicuculline revealed underlying EPSPs (latency, 7.8 +/- 0.8 ms; mean Vr, -8 +/- 10 mV) with two components: (a) fast (rise and half-decay times of 5.83 +/- 1.3 ms and 19 +/- 4.4 ms respectively), with reversible blockade by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX); (b) slow (4- to 5-fold increase in rise and half-decay time), with reversible reduction by (-)-aminophosphonovaleric acid (APV). 3. During 10 Hz stimulation, EPSPs summated into 3-7 mV depolarizing envelopes lasting 1.5-3.0 s and sustaining action potential bursts. Depolarizing envelopes displayed voltage dependence, and were enhanced after removal of extracellular magnesium, diminished by APV and completely abolished by APV and CNQX together. 4. Thus, non-NMDA receptors probably mediate fast EPSPs whereas NMDA receptors mediate slow EPSPs and depolarizing envelopes. OVLT-evoked EPSPs were only seen in vasopressin-immunoreactive neurones. 5. These observations indicate converging inhibitory and target-selective excitatory amino acid-mediated inputs from OVLT to SON; the latter may modulate the excitability of SON vasopressin neurones to a hyperosmotic challenge.

Depolarizing action of secretory granule protein 7B2 on rat supraoptic neurosecretory neurons.

A novel precursor neuropeptide termed 7B2 is present within specific brain areas, including the hypothalamic magnocellular neurosecretory neurons, and appears to be processed to smaller fragments. In order to determine whether specific C-terminal fragments of 7B2 might exert local effects on neurosecretory cells, we used intracellular current-clamp recordings in supraoptic neurons maintained in superfused hypothalamic explants to evaluate membrane potential and resistance changes in 25 supraoptic nucleus neurons during bolus applications of 7B2 174-186 and two other C-terminal peptide fragments 7B2 156-173 and 7B2 141-150. In 15 supraoptic neurons, only the 7B2 174-186 fragment induced a gradual 2-8 mV membrane depolarization that lasted for 4 to 30 min and was accompanied by 15+/-8% reduction in input resistance. Immunocytochemical identification of the recorded cells revealed that both vasopressin (VP)- and oxytocin (OT)-containing neurons were depolarized by 7B2 174-186. These data suggest that 7B2 174-186 is a biologically active fragment of 7B2 and may regulate the excitability of magnocellular supraoptic nucleus neurons.

Visualization of outgrowing axons of grafted neurons by anterograde labelling with Phaseolus vulgaris leucoagglutinin in the motor cortex of the rat.

Fetal cerebral cortical tissue was transplanted into an aspirated lesion cavity made in the sensorimotor cortex of adult rats. Ten weeks after grafting, outgrowing fibers from the graft were visualized by an anterograde tracing technique using Phaseolus vulgaris leucoagglutinin (PHA-L). It was demonstrated that the efferent fibers grew into the neighboring host cortical tissue, the corpus callosum and in some cases approached caudate/putamen. Characteristic axon arborization with abundant boutons were found in the host cortical tissue, but only in close vicinity to the grafts. It is concluded that the PHA-L anterograde tracing technique can be a useful tool to assess the degree of anatomical integration of the transplants into the host tissue.