Postnatal maturational properties of rat parafascicular thalamic neurons recorded in vitro.

Thalamic relay neurons have homogeneous, adult-like firing properties and similar morphology by 12 days postnatally (PN 12). Parafascicular (Pf) neurons have a different morphology compared with typical thalamic relay neurons, but the development of their electrophysiological properties is not well studied. Intracellular recordings in PN 12-50 Pf neurons revealed several heterogeneous firing patterns different from those in thalamic relay neurons. Two types of cells were identified: Type I cells displayed a fast afterhyperpolarization (AHP) followed by a large-amplitude, slow AHP; whereas Type II cells had only a fast AHP. These cell types had overlapping membrane properties but differences in excitability. Some properties of Pf neurons were adult-like by PN 12, but, unlike thalamic relay neurons, there were significant maturational changes thereafter, including decreased action potential (AP) duration, increased fast AHP amplitude and increased excitability. Pf neurons did not exhibit rhythmic bursting and generally lacked low-threshold spike (LTS) responses that characterize thalamic relay neurons. Pf neurons exhibited nonlinear I-V relationships, and only a third of the cells expressed the time and voltage-dependent hyperpolarization activated (Ih) current, which declined with age. These results indicate that the morphological differences between Pf neurons and typical thalamic relay neurons are paralleled by electrophysiological differences, and that Pf membrane properties change during postnatal development.

Intracellular recording of lamina X neurons in a horizontal slice preparation of rat lumbar spinal cord.

A horizontal slice preparation of postnatal rat lumbar spinal cord has been developed which allows correlative observations of the morphology, electrophysiology, and receptor pharmacology of lamina X neurons. These slices better maintain afferent input and somatodendritic morphology and are amenable to subsequent immunohistochemical processing. Stable intracellular recordings obtained from postnatal day 14-45 animals reveal that a number of different intrinsic membrane conductances contribute to the regulation of excitability in lamina X neurons. In addition, lamina X neurons possess inhibitory GABAergic as well as excitatory glutamate and cholecystokinin receptors. This preparation will be useful in future studies designed to characterize developmental changes in the intrinsic membrane properties, synaptic profiles and neuropeptide responsiveness of lamina X neurons in the rat. Such a characterization is important given that lamina X represents a unique sexually dimorphic region that is a convergence site for somatic and visceral afferent inputs, which includes nociceptive information.

Sex differences in the response of postnatal rat lumbar lamina X neurons to exogenously applied galanin recorded in vitro.

Intracellular recording techniques were used in a horizontal slice preparation of postnatal rat lumbar spinal cord to compare the responses of male and female lamina X neurons to exogenously applied galanin. Although there was no significant sex difference in the resting membrane potential or input resistance of neurons, superfusion of galanin 1-16 (1 microM) produced a membrane hyperpolarization that averaged -5.3 mV in males and only -2.0 mV in females. The galanin-induced membrane hyperpolarization of lamina X neurons was accompanied by an inconsistent and varied change in input resistance. No depolarizing effect of galanin was detected in either sex. Galanin did not significantly alter the spike shape, amplitude, after hyperpolarization or locally evoked synaptic responses. The more than 2.5 fold significant sex difference in response to galanin occurred at a developmental timepoint at which lamina X expressed a comparably higher amount of galanin-like immunoreactivity in males compared to females. These results provide the first indication of a sex difference in the response of lamina X neurons to any neuropeptide. Given the antinociceptive role of galanin in the spinal cord, these results raise the possibility for the presence of distinct physiological and anatomical substrates for sex-dependent differences in nociceptive processing in lamina X of the lumbosacral spinal cord.

Heterogeneity of astroglia cultured from adult human temporal lobe.

This study characterized the morphological and electrophysiological diversity of astroglia cultured from adult human cerebral temporal lobe, and explored the influence of the cytokine interleukin-1beta on these cells. The cultures contained astroglia positive for glial fibrillary acidic protein which were flat, bipolar or multipolar in shape and variable in size. A subpopulation of the bipolar and multipolar cells was positive for S100 protein. The most striking feature of these cultures was the presence of glia with long (600 micrometer) processes with few branches or only terminal branches. Patch clamp recordings of the non-stellate process bearing cells revealed prominent inward Na(+) and transient and sustained outward K(+) conductances. Distinct differences in the relative proportion of these conductances were evident among cells but did not appear to be correlated with cell morphology. Treatment of cultures with interleukin-1beta for 96 h did not change total protein content, but increased the content of S100beta protein and decreased the content of glial fibrillary acidic protein. The findings indicate that cultures of adult human cerebrum contain subpopulations of morphologically and electrophysiologically pleomorphic glial fibrillary acidic protein positive astroglia, exhibit increased levels of the neurotrophic factor S100beta when exposed to interleukin-1beta, and may serve as a useful model for investigation of glial involvement in neuropathology.

N-ethylmaleimide selectively blocks presynaptic GABA-B autoreceptor but not heteroreceptor-mediated inhibition in adult rat striatal slices.

Intracellular recording techniques were used in slices of adult rat striatum to compare the sensitivity of presynaptic gamma-aminobutyric acid type B (GABA-B) autoreceptor and heteroreceptor-mediated inhibition to the sulfhydryl alkylating agent, N-ethylmaleimide (NEM). NEM (100 microM) alone had no significant effect on resting potential or input resistance and did not consistently effect stimulus-evoked responses. Treatment of slices with NEM for up to 1 h, either in the presence or the absence of the GABA-B receptor-specific agonist, baclofen (BAC; 100 microM), completely blocked the BAC-induced depression of inhibitory, but not excitatory, postsynaptic potentials. This differential sensitivity to NEM alkylation suggests that in the adult rat striatum, the presynaptic GABA-B autoreceptors and heteroreceptors may exhibit distinct receptor-effector coupling mechanisms.

Neurotoxicity of 25-OH-cholesterol on sympathetic neurons.

Cultured rat sympathetic neurons derived from postnatal rat superior cervical ganglia (SCG) were used to compare the neurotoxicity of several cholesterol oxides. The cholesterol oxides tested included: 7-beta-OH-, 7-keto-, 19-OH-, 22(R)-OH-, 22(S)-OH-, and 25-OH-cholesterol. These agents caused an acute as well as a delayed toxicity in sympathetic neurons with 25-OH-cholesterol appearing to be the most toxic. A time-dependent experiment indicated that 25-OH-cholesterol at 4 microg/ml (10 microM) was able to kill 50% of the cells in 36 h. Morphological studies indicate that most of the cells do not exhibit a structural change similar to that observed in neuronal programmed cell death. Whole-cell patch clamp recording of untreated controls and 25-OH-cholesterol (2 microg/ml)-treated cells indicated that this toxicity was not accompanied by significant changes in voltage-dependent calcium channel activity. A number of pharmacological agents including ethylene glycolbis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA), cycloheximide, KCl, vitamin E, and methyl-beta-cyclodextrin were able to prevent the 25-OH-cholesterol-induced cell death to various degrees. These results suggest that, in addition to causing pathological changes in cells directly involved in atherosclerosis, cholesterol oxides may induce neurotoxicity in sympathetic neurons.

Neurotoxicity of 25-OH-cholesterol on NGF-differentiated PC12 cells.

PC12 cells induced to differentiate with nerve growth factor were used to study the neurotoxicity of 25-OH-cholesterol. This agent induced a dose- and time-dependent cell death in neuronal PC12 cells. Cells treated with this agent showed condensed nuclei, a morphology similar to that of cells dying of programmed cell death. However, agents known to prevent neuronal programmed cell death (cyclic AMP, KCl, aurintricarboxylic acid, and cycloheximide) failed to prevent the 25-OH-cholesterol-mediated cytotoxicity. On the other hand, cell death induced by 25-OH-cholesterol was prevented by treatment with vitamin E and methyl-beta-cyclodextrin. In contrast to observations made in other cell types, whole-cell patch clamp recording of neuronal PC12 cells revealed that treatment with 25-OH-cholesterol did not significantly alter calcium influx through voltage-dependent channels. These results provide the first characterization of the toxicity of cholesterol oxides toward neuronal PC12 cells, which should be useful in future studies on the interactions between cholesterol oxides and cells from the nervous system.

Establishment of two morphologically distinct PC12 cell lines resistant to 25-OH-cholesterol toxicity.

Two novel populations of spontaneous PC12 cell mutants resistant to a toxic concentration of 25-OH-cholesterol (5 microg/ml, 12.5 microM) were isolated and designated as R25R and F25R based on cell morphology. R25R consisted of round cells that were morphologically similar to the parent PC12 cells, and responded to nerve growth factor by extending neurites. F25R was a group of process-bearing flat cells that did not assume a neuronal morphology in the presence of nerve growth factor. These two cell lines also acquired some cross-resistance toward other cholesterol oxides. Nerve growth factor induced prominent voltage-dependent calcium currents in parent PC12 cells and in R25R, but not in F25R. Further experiments indicated that the parent PC12 cells, R25R and F25R exhibited different properties when challenged with a variety of toxic insults, including amphotericin B, serum withdrawal and beta-amyloid protein treatment.

Acute exposure to 25-hydroxy-cholesterol selectively reduces GABAb and not GABAa receptor-mediated synaptic inhibition.

Intracellular recording techniques were used to study the effects of the cholesterol oxide, 25-hydroxycholesterol (25-OH-Chol), on gamma-aminobutyric acid (GABA) receptor-mediated inhibitory postsynaptic potentials (IPSPs) in brain slices of the rat lateral septum. Superfusion of 25-OH-Chol increased the peak amplitude of the GABAa IPSP in more than half of the neurons tested, many of which exhibited a similar increase in the GABAb IPSP. However, some neurons exhibited a gradual decrease in input resistance and a selective reduction or blockade of the GABAb IPSP during prolonged exposure. Cholesterol partly mimicked the effects of 25-OH-Chol. These findings indicate that 25-OH-Chol can selectively reduce or block metabotropic GABAb while sparing ionotropic GABAa receptor-mediated synaptic inhibition. Our results indicate that brain slices can be used to study the effects of short term alterations in cholesterol on the excitability and synaptic integration properties of neurons.

Developmental changes in serotonergic receptor-mediated modulation of embryonic chick motoneurons in vitro.

Intracellular recordings were obtained from antidromically identified motoneurons in an embryonic chick spinal cord slice preparation at two developmental stages (embryonic days 12 and 18, E12 and E18) which bracket a critical period in spinal cord growth. The resting membrane potential of chick motoneurons did not change significantly between E12 and E18, but there was a significant decrease in neuronal input resistance. A small inward rectification was present in cells of both ages, although a lower proportion of E12 motoneurons exhibited inward rectification compared to E18 motoneurons. Injection of depolarizing current pulses revealed that most E12 motoneurons exhibited spike adaptation, while the majority of E18 motoneurons showed high frequency tonic firing. Bath application of serotonin (5-HT) and its agonists 5-carboxamido-tryptamine (5-CT, a 5-HT1 agonist) and alpha-methyl 5-HT (a 5-HT2 agonist) produced hyperpolarizing responses accompanied by decreased input resistance in all E12 motoneurons studied. The same three agonists produced depolarizing responses and increased input resistance in all E18 motoneurons studied. The effects of serotonergic agonists on motoneuronal excitability were tested using depolarizing current pulses. In most cases, serotonergic agonists caused a decrease in firing frequency during the hyperpolarizing response in E12 neurons. At E18, bath application of 5-HT, 5-CT or alpha-methyl 5-HT produced an increase in firing frequency in all motoneurons during the depolarizing response. Our results indicate that both 5-HT1 and 5-HT2 receptor subtypes contribute to modulation of chick motoneuron excitability and appear to reverse the polarity of their effects on membrane potential after a critical period in development of the spinal cord.

Transforming growth factor-beta2 selectively alters the developmental expression of the fast transient A-current in cultured rat superior cervical ganglion neurons.

Cultures of neonatal rat superior cervical ganglion (SCG) were utilized to examine the ability of transforming growth factor-beta2 (TGFbeta2) to alter voltage-gated K+ channel development. Whole-cell patch clamp recordings were used to monitor changes in three separate K+ currents: A rapidly inactivating A-current (I(Af)), a slowly inactivating A-current (I(As)), and a non-inactivating current (I(K)). Continuous TGFbeta2 (10 ng/ml) treatment selectively altered the normal developmental decrease in I(Af) expression in SCG neurons, but did not significantly change I(As) or I(K) expression. After 2 weeks of treatment, the mean I(Af) current density in control cultures had decreased 67%, while the I(Af) current density in TGFbeta2 treated cultures remained near initial values (approximately 2.7-fold higher than control). This difference remained even after 4 weeks of exposure. TGFbeta2 did not appear to change the activation kinetics or voltage-dependence of I(Af). These findings indicate that TGFbeta2 may play an important role in modulating the development of neuronal excitability by regulating the expression of voltage-gated K+ channels.

Developmental changes in the effects of serotonin and N-methyl-D-aspartate on intrinsic membrane properties of embryonic chick motoneurons.

A spinal cord slice preparation was developed in order to study developmental changes in intrinsic membrane properties and in responses to N-methyl-D-aspartate and serotonin in embryonic chick motoneurons. Transverse spinal cord slices were obtained from chick embryos over a series of developmental stages (embryonic days 12-18). Intracellular recordings were obtained from 87 antidromically identified motoneurons. During the stages examined, the average resting membrane potential did not vary significantly, the voltage threshold of current-evoked action potentials became significantly more negative, there was a non-significant trend towards a decrease in the recorded input resistance, but there were no significant changes observed in the membrane time constant. There were significant developmental changes in the waveform of the current-evoked action potentials. The average amplitude of the action potentials increased over the stages studied, while the action potential duration measured at half-amplitude decreased. All of the motoneurons examined were maximally depolarized by bath application of 50 microM N-methyl-D-aspartate. The depolarization persisted in the presence of tetrodotoxin but was blocked by 100 microM 2-amino-5-phosphonopentanoic acid and, therefore, was at least partially due to a direct action of N-methyl-D-aspartate on motoneuronal receptors. The average amplitude of the N-methyl-D-aspartate-induced depolarizations decreased significantly over the stages examined. In contrast, bath application of 50 microM serotonin produced either depolarizing or hyperpolarizing responses depending on the developmental age of the motoneuron. Serotonin induced a depolarization in about 50% of the motoneurons at embryonic day 12, 69% of the motoneurons at embryonic day 15 and 100% of the motoneurons recorded from at embryonic day 18. These findings reveal important developmental changes in intrinsic membrane responses and action potential properties of chick motoneurons recorded from a slice preparation. We have also documented changes in the motoneuronal responses to serotonin, a neurotransmitter used by a major descending projection, and N-methyl-D-aspartate, which activates glutamate receptors known to contribute to synaptic activity in segmental circuits.

Transforming growth factor-beta 2 both stimulates and inhibits neurogenesis of rat cerebellar granule cells in culture.

Transforming growth factor-beta 2 (TGF beta 2) is expressed in the developing cerebellar cortex during the period of granule cell proliferation and maturation. However, the role of TGF beta 2 in granule cell development is confused by conflicting observations regarding TGF beta 2 control of neurogenesis. To resolve these conflicts and determine the effect of TGF beta 2 on neurogenesis, rat cerebellar granule cell cultures were treated with TGF beta 2 (0.1-100 ng/ml, 24 h) in the presence or absence of exogenous serum. Neuroblast proliferation was quantified by bromodeoxyuridine and [3H]thymidine incorporation. TGF beta 2 stimulated proliferation to 220% of controls in the presence of serum (ED50 = 0.4 ng/ml) based on bromodeoxyuridine labeled granule cell counts. In contrast, in serum free medium, TGF beta 2 inhibited proliferation 75% (ED50 = 0.7 ng/ml). DNA synthesis measured by [3H]thymidine incorporation was increased to 122% in the presence of serum factors, but inhibited 70% in serum free medium, as a result of TGF beta 2 activity. Thus, TGF beta 2 differentially regulates neurogenesis of cerebellar granule cells depending on the presence of exogenous, undefined regulatory factors derived from serum. This suggests that TGF beta 2 activity in cerebellar neurogenesis is complex as it may be modulated by the repertoire of other endogenous regulatory factors in the developing cerebellar cortex.

Retrograde labeling of rat dorsolateral septal nucleus neurons following intraseptal injections of WGA-HRP.

Projection neurons in the rat dorsolateral septal nucleus (DLSN) were retrogradely labeled following intraseptal injection of wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP). Injections of WGA-HRP centered in the medial septum (MS) and parts of the intermediate and ventrolateral subdivisions of the lateral septum retrogradely labeled only a few centrally scattered multipolar-shaped neurons. In contrast, injections placed in the nucleus of the diagonal band of Broca (DBB) consistently resulted in labeling of DLSN neurons of all sizes and shapes. Large injections in rostral DBB appeared to retrogradely label every DLSN neuron, while similar injections in caudal DBB only labeled neurons in restricted regions of the nucleus. A collection of small cells forming the ventricular border of caudal DLSN and a group of larger cells situated in the dorsolateral tip of rostral DLSN were consistently labeled following each DBB injection. The pattern of retrogradely labeled neurons in the DLSN appeared in a complementary fashion to that seen in the other lateral septal nuclei. Our findings support the conclusion that the DLSN is a morphologically heterogeneous nucleus consisting almost entirely of projection neurons. The pattern of retrograde labeling in the lateral septum suggests that these projection neurons may be topographically organized since distinct subpopulations of cells were labeled following different injections in the MS/DBB complex.

Transforming growth factor-beta2 increases NMDA receptor-mediated excitotoxicity in rat cerebral cortical neurons independently of glia.

The ability of transforming growth factor-beta2 (TGFbeta2) to directly regulate neuronal sensitivity to glutamate and N-methyl-D-aspartate (NMDA) excitotoxicity in rat cerebral cortical neurons was investigated. Mixed neuronal-glial cultures treated with TGFbeta2 (1-10 ng/ml) exhibited a significant 25-50% increase in neuronal death compared to control cultures. TGFbeta2 potentiation of this endogenous glutamate excitotoxicity was blocked by the selective NMDA receptor antagonist, 2-amino-5-phosphonovalerate. In addition, neuronal death induced by brief NMDA exposure in both mixed neuronal-glial and pure neuronal cultures was increased by TGFbeta2 (1-30 ng/ml) with a similar dose-response curve. These findings indicate that TGFbeta2, at physiologically relevant concentrations, potentiates NMDA receptor-mediated excitotoxicity and that this occurs independently of TGFbeta2 effects on glia.

A novel tetrodotoxin-resistant sodium current from an immortalized neuroepithelial cell line.

1. Voltage-gated ionic currents were recorded from cells of an immortalized neuroepithelial cell line named V1. The cell line was produced by insertion of the temperature-sensitive tsA58 allele of the SV40 large T-antigen into embryonic day 14 mouse hypothalamic cells. V1 cells display a mixed immature neural-glial phenotype and have two phenotypes, round and flat. 2. Recordings from round V1 cells demonstrate voltage-gated Na+ and K+ currents (n = 297), while no voltage-gated currents were observed in flat V1 cells (n = 45). Voltage-gated currents were recorded from cells cultured at both permissive and restrictive temperatures. 3. Internal Cs+ and external tetraethylammonium (TEA) were used to suppress outward currents. The remaining inward current has rapid activation and inactivation time constants which decreased as the test potential increased. In different cells, the amplitude of the peak inward current varied from about 50 pA to as large as 4500 pA (in 120 mM external Na+). The reversal potential for the inward current was close to the predicted Nernst equilibrium potential for Na+. Both the magnitude and reversal potential of the inward current were dependent on the external Na+ concentration. It is therefore considered to be a Na+ current, INa. 4. INa was found to be TTX resistant. About 5% of the INa was blocked by 200 nM TTX and 20 microM TTX fully suppressed the Na+ current. The apparent Kd for TTX blockade was estimated to be 1.49 microM. 5. The activation kinetics of INa could be described by a Hodgkin-Huxley model with an m3 variable. The time constants of activation and inactivation of INa are fast, similar to those of the TTX-resistant and TTX-sensitive Na+ currents in central nervous system neurons and glial cells. 6. The divalent and trivalent cations Cd2+, Co2+, Ni2+, Zn2+ and La3+ shifted the activation of INa to more positive potentials and decreased the maximal conductance in a dose-dependent manner. The apparent Kd values for blockade of the INa by Cd2+, Co2+, Ni2+, Zn2+ and La3+ were 430, 3500, 1900, 83 and 202 microM, respectively. 7. The addition of phorbol myristate acetate, an activator of protein kinase C, consistently produced a reduction in the amplitudeof INa without affecting the time course of activation or inactivation. 8. INa in V1 cells expresses a unique combination of voltage and time kinetics and sensitivity to blockade by TTX and cations. We hypothesize that this Na+ current may be expressed transiently during development of the central nervous system at the stage of development represented by the V1 cell line.

Morphological and electrophysiological evidence for electrotonic coupling of rat dorsolateral septal nucleus neurons in vitro.

Intracellular injections of Lucifer Yellow were utilized to evaluate the incidence of dye-coupling among dorsolateral septal nucleus (DLSN) neurons recorded from slice preparations of adult rat septal nuclei. Twenty percent of single injections of Lucifer Yellow resulted in pairs of labeled neurons. These dye-coupled cells were morphologically heterogeneous and did not exhibit any morphological characteristics that could be used to distinguish them from non dye-coupled neurons. The spatial separation of cell bodies and close apposition of dendrites within each pair indicated that the dye transfer site(s) were situated at dendrodendritic and/or dendrosomatic rather than somatosomatic junctions. The main axon of some dye-coupled neurons gave rise to intrinsic axon collaterals prior to exiting the nucleus indicating that these coupled neurons function as projection neurons as well as local circuit interneurons. Electrophysiological recordings of the passive membrane properties and spontaneous activity of individual dye-coupled neurons revealed no significant difference from non dye-coupled cells in the DLSN. Some neurons exhibited spontaneously occurring fast potentials which presumably represent electrotonic potentials. These fast potentials were often tightly coupled with action potentials but could be distinguished from synaptic potentials by their shape and their lack of voltage-dependent changes in amplitude. These morphological and supportive electrophysiological data provide the first indirect evidence for electrotonic coupling of dorsolateral septal neurons. The functional significance of this coupling may lie in the potential for synchronization of the output of the DLSN which could play an important role in the septal maintenance and modulation of hippocampal Theta rhythm.

Direct muscarinic and nicotinic receptor-mediated excitation of rat medial vestibular nucleus neurons in vitro.

We have utilized intracellular recording techniques to investigate the cholinoceptivity of rat medial vestibular nucleus (MVN) neurons in a submerged brain slice preparation. Exogenous application of the mixed cholinergic agonists, acetylcholine (ACh) or carbachol (CCh), produced predominantly membrane depolarization, induction of action potential firing, and decreased input resistance. Application of the selective muscarinic receptor agonist muscarine (MUSC), or the selective nicotinic receptor agonists nicotine (NIC) or 1,1-dimethyl-4-phenylpiperazinium (DMPP) also produced membrane depolarizations. The MUSC-induced depolarization was accompanied by decreased conductance, while an increase in conductance appeared to underlie the NIC- and DMPP-induced depolarizations. The muscarinic and nicotinic receptor mediated depolarizations persisted in tetrodotoxin and/or low Ca2+/high Mg2+ containing media, suggesting direct postsynaptic receptor activation. The MUSC-induced depolarization could be reversibly blocked by the selective muscarinic-receptor antagonist, atropine, while the DMPP-induced depolarization could be reversibly suppressed by the selective ganglionic nicotinic-receptor antagonist, mecamylamine. Some neurons exhibited a transient membrane hyperpolarization during the depolarizing response to CCh or MUSC application. This transient inhibition could be reversibly blocked by the gamma-aminobutyric acid (GABA) antagonist, bicuculline, suggesting that the underlying hyperpolarization results indirectly from the endogenous release of GABA acting at GABA receptors. This study confirms the cholinoceptivity of MVN neurons and establishes that individual MVN cells possess muscarinic as well as nicotinic receptors. The data provide support for a prominent role of cholinergic mechanisms in the direct and indirect regulation of the excitability of MVN neurons.

Spontaneous bursting and non-bursting activity in morphologically identified neurons of the rat dorsolateral septal nucleus, in vitro.

Membrane potential-dependent changes in the repetitive firing properties of morphologically identified rat dorsolateral septal nucleus neurons were investigated in a submerged slice preparation using intracellular recording techniques and lithium acetate-Lucifer Yellow-filled microelectrodes. The results indicate that the majority of dorsolateral septal nucleus neurons are capable of burst firing and suggest, moreover, the existence of neuronal subtypes with distinct differences in spike waveform and the pattern of spontaneous activity. In the largest proportion of neurons, single spike activity predominated at membrane potentials near rest while burst-like discharges prevailed at more hyperpolarized membrane potentials. Less frequently observed were neurons exhibiting different burst waveforms at various membrane potentials. In a few neurons, hyperpolarization slowed neuronal firing but did not elicit burst-like discharges. Characteristics such as the presence of burst or single spike discharges, spike afterpotentials, and the membrane potential dependence of repetitive firing patterns did not appear to be closely associated with membrane time constant, membrane resistance, or resting membrane potential. A detailed examination of the somatodendritic and axonal morphology of the Lucifer Yellow-filled cells revealed that these electrophysiologically identified neurons in the dorsolateral septal nucleus are morphologically heterogeneous. However, there did not appear to be any correlation between a particular somatodendritic morphology and the expression of a distinct spontaneous firing pattern. The present findings demonstrate that neurons in the rat dorsolateral septal nucleus are morphologically diverse and capable of intrinsically generating rhythmic neuronal activity. Similar patterns of rhythmic neuronal firing in vivo may provide a substrate for the integration of afferent neuronal activity and have a central role in intraseptal circuitry necessary for generation of hippocampal theta rhythm.