Changes in dendritic axial resistance alter synaptic integration in cerebellar Purkinje cells

The ability of neurons to process synaptic inputs depends critically on their passive electrical properties. The intracellular resistivity, R_i, is one of the parameters that determine passive properties, yet few experiments have explored how changes in R_i might affect synaptic integration. In this work, I addressed this issue by using targeted dendritic occlusion to locally increase R_i in cerebellar Purkinje cells and examining the consequences of this manipulation for the summation of synaptic inputs. To achieve dendritic occlusion, I used two glass micropipettes to gently pinch the dendritic trunk close to the soma. This pinching produced stereotypical changes in the responses to test pulses applied at the soma under voltage and current clamp. A simple model confirmed that these changes were due to increases in R_i in the dendritic trunk. These localized increases in R_i produced striking alterations in the shapes of postsynaptic potentials at the soma, increasing their amplitude and accelerating their decay kinetics. As a consequence, dendritic occlusion sharpened temporal precision during the summation of synaptic inputs. These findings highlight the importance of local changes in intracellular resistivity for the passive electrical properties of neurons, with implications for their ability to process synaptic information.     

The potassium A-current,low firing rates and rebound excitation in Hodgkin-Huxley models

It is widely believed, following the work of Connor and Stevens (1971,J. Physiol. Lond. 214, 31–53) that the ability to fire action potentials over a wide frequency range, especially down to very low rates, is due to the transient, potassium A-current (I _A). Using a reduction of the classical Hodgkin-Huxley model, we study the effects ofI _A on steady firing rate, especially in the near-threshold regime for the onset of firing. A minimum firing rate of zero corresponds to a homoclinic bifurcation of periodic solutions at a critical level of stimulating current. It requires that the membrane's steady-state current-voltage relation be N-shaped rather than monotonic. For experimentally based genericI _A parameters, the model does not fire at arbitrarily low rates, although it can for the more atypicalI _A parameters given by Connor and Stevens for the crab axon. When theI _A inactivation rate is slow, we find that the transient potassium current can mediate more complex firing patterns, such as periodic bursting in some parameter regimes. The number of spikes per burst increases asg _A decreases and as inactivation rate decreases. We also study howI _A affects properties of transient voltage responses, such as threshold and firing latency for anodal break excitation. We provide mathematical explanations for several of these dynamic behaviors using bifurcation theory and averaging methods.     

In vivo analysis of inhibitory synaptic inputs and rebounds in deep cerebellar nuclear neurons

Neuronal function depends on the properties of the synaptic inputs the neuron receive and on its intrinsic responsive properties. However, the conditions for synaptic integration and activation of intrinsic responses may to a large extent depend on the level of background synaptic input. In this respect, the deep cerebellar nuclear (DCN) neurons are of particular interest: they feature a massive background synaptic input and an intrinsic, postinhibitory rebound depolarization with profound effects on the synaptic integration. Using in vivo whole cell patch clamp recordings from DCN cells in the cat, we find that the background of Purkinje cell input provides a tonic inhibitory synaptic noise in the DCN cell. Under these conditions, individual Purkinje cells appear to have a near negligible influence on the DCN cell and clear-cut rebounds are difficult to induce. Peripheral input that drives the simple spike output of the afferent PCs to the DCN cell generates a relatively strong DCN cell inhibition, but do not induce rebounds. In contrast, synchronized climbing fiber activation, which leads to a synchronized input from a large number of Purkinje cells, can induce profound rebound responses. In light of what is known about climbing fiber activation under behaviour, the present findings suggest that DCN cell rebound responses may be an unusual event. Our results also suggest that cortical modulation of DCN cell output require a substantial co-modulation of a large proportion of the PCs that innervate the cell, which is a possible rationale for the existence of the cerebellar microcomplex.     

Thin dendrites of cerebellar interneurons confer sublinear synaptic integration and a gradient of short-term plasticity

Interneurons are critical for neuronal circuit function, but how their dendritic morphologies and membrane properties influence information flow within neuronal circuits is largely unknown. We studied the spatiotemporal profile of synaptic integration and short-term plasticity in dendrites of mature cerebellar stellate cells by combining two-photon guided electrical stimulation, glutamate uncaging, electron microscopy, and modeling. Synaptic activation within thin (0.4?μm) dendrites produced somatic responses that became smaller and slower with increasing distance from the soma, sublinear subthreshold input-output relationships, and a somatodendritic gradient of short-term plasticity. Unlike most studies showing that neurons employ active dendritic mechanisms, we found that passive cable properties of thin dendrites determine the sublinear integration and plasticity gradient, which both result from large?dendritic depolarizations that reduce synaptic driving force. These integrative properties allow stellate cells to act as spatiotemporal filters of synaptic input patterns, thereby biasing their output in favor of sparse presynaptic activity.     

Effect of harmaline on firing pattern of rat cerebellar Purkinje cells in ontogenesis

In this work, responses of rat Purkinje cells to intraperitoneal administration of the hallucinogenic alkaloid harmaline (0.15 mg/kg) were studied in the course of ontogenesis. The experiments were carried out on Wistar rats of three age groups: rat pups (13–18 days), adult animals (2–7 months), and aged rats (25–36 months). In Purkinje cell firings, two types of electric reactions were revealed; they were similar in all age group of the animals. In cells with the 1st type of reactions, in response to the harmaline administration there was recorded a significant increase of frequency of complex spikes, accompanied by disappearance of simple spikes. In the activity of Purkinje cells of the 2nd type, the complex spike frequency also increased; however, the firing simple spikes were preserved, although with a decrease of their frequency as compared with norm. Essential changes of activity of the cerebellar Purkinje cells were found in the rat pups and aged animals in comparison with adult rats, which agrees well with immaturity of various cerebellar structures in the first case and with involutionary changes in the second case.     

Effect of harmaline on firing pattern of rat cerebellar Purkinje cells in ontogenesis

In this work, responses of rat Purkinje cells to intraperitoneal administration of the hallucinogenic alkaloid harmaline (0.15 mg/kg) were studied in the course of ontogenesis. The experiments were carried out on Wistar rats of three age groups: rat pups (13-18 days), adult animals (2-7 months), and aged rats (25-36 months). In Purkinje cell firings, two types of electric reactions were revealed; they were similar in all age group of the animals. In cells with the 1st type of reactions, in response to the harmaline administration there was recorded a significant increase of frequency of complex spikes, accompanied by disappearance of simple spikes. In the activity of Purkinje cells of the 2nd type, the complex spike frequency also increased; however, the firing simple spikes were preserved, although with a decrease of their frequency as compared with norm. Essential changes of activity of the cerebellar Purkinje cells were found in the rat pups and aged animals in comparison with adult rats, which agrees well with immaturity of various cerebellar structures in the first case and with involutionary changes in the second case.     

Assessment of spatial habitat heterogeneity by coupling data-driven habitat suitability models with a 2D hydrodynamic model in small-scale streams

Habitat assessment considering habitat quality and quantity is a key approach in conservation and restoration works for biodiversity and ecosystems. In this regard, application of hydrodynamic model for modeling instream habitat conditions and machine learning (ML) methods for modeling habitat suitability of a target species can contribute to better modeling practices in ecohydraulics. Despite the importance of small streams for aquatic ecosystems, previous studies in ecohydraulics have been conducted mainly in medium to large rivers, often disregarding small-scale streams such as agricultural canals. The aim of this study is to demonstrate the applicability of a coupled use of ML and a two-dimensional (2D) hydrodynamic model for assessing spatial habitat heterogeneity in small-scale agricultural canals in Japan. Using abundance data of Japanese medaka (Oryzias latipes), four ML methods, namely artificial neural networks (ANNs), classification and regression trees (CARTs), random forests (RF) and support vector machines (SVMs), were applied to develop habitat suitability models considering water depth and flow velocity. A 2D hydrodynamic model was developed based on field surveys in two types of agricultural canals, namely earthen and concrete-lined canals. Information entropy was used for assessing the spatial heterogeneity of instream habitat conditions. As a result, the hydrodynamic models could model instream habitat conditions in a reasonable accuracy. Despite the differences in accuracies in habitat modeling, the four ML methods illustrated similar habitat suitability information for Japanese medaka. The coupled ecohydraulics modeling approach could quantify habitat quality and its spatial heterogeneity, based on which the differences between the earthen and concrete-lined canals were quantitatively assessed. This study demonstrated the applicability of ML-based habitat suitability evaluation and a 2D hydrodynamic model for modeling the spatial distribution of habitat suitability and assessing its spatial heterogeneity. Further study, assessing the spatial heterogeneity in various types of flows including natural/artificial and small/large streams, can contribute to establish quantitative criteria for an ecologically sound habitat and improved ecofriendly construction works in small-scale rivers and streams.     

Presence of protein I, a phosphoprotein associated with synaptic vesicles, in cerebellar granule cells

Abstract: The cerebellar levels of Protein I, a synapse-specific neuronal phosphoprotein, have been investigated in the cerebellar mouse mutants staggerer ( sg ), weaver ( wv ), nervous ( nr ), and Purkinje cell degeneration ( pcd ). The Protein I concentration was reduced by about 66% in sg and wv mutants, representing a 90% loss of Protein I per cerebellum. A heterozygote effect was observed in the wv mutant. These results indicate that a great majority of Protein I in the normal cerebellum may be present in the granule cells. in nr mutants the cerebellar Protein I concentration was reduced by only 12% in 62-day-old mice, suggesting that Purkinje cells contribute little to cerebellar Protein I. However, a greater reduction was observed in pcd mutants, which may reflect on the nature of the pcd mutation.     

Pre and post synaptic NMDA effects targeting Purkinje cells in the mouse cerebellar cortex

N-methyl-D-aspartate (NMDA) receptors are associated with many forms of synaptic plasticity. Their expression level and subunit composition undergo developmental changes in several brain regions. In the mouse cerebellum, beside a developmental switch between NR2B and NR2A/C subunits in granule cells, functional postsynaptic NMDA receptors are seen in Purkinje cells of neonate and adult but not juvenile rat and mice. A presynaptic effect of NMDA on GABA release by cerebellar interneurons was identified recently. Nevertheless whereas NMDA receptor subunits are detected on parallel fiber terminals, a presynaptic effect of NMDA on spontaneous release of glutamate has not been demonstrated. Using mouse cerebellar cultures and patch-clamp recordings we show that NMDA facilitates glutamate release onto Purkinje cells in young cultures via a presynaptic mechanism, whereas NMDA activates extrasynaptic receptors in Purkinje cells recorded in old cultures. The presynaptic effect of NMDA on glutamate release is also observed in Purkinje cells recorded in acute slices prepared from juvenile but not from adult mice and requires a specific protocol of NMDA application.     

Effects of long-term hypothyroidism in the morphology and synaptic organization of cerebellar ectopic granule cells

Abundant ectopic granule cells scattered in the cerebellar molecular layer have been observed in 30-day-old hypothyroid rats. Their morphological features indicate that they must be regarded as mature heterotopic cells arrested during their migration towards the granular layer. As their impoverished dendritic trees are identical to those seen in controls, it is unlikely that the lack of thyroid hormones played a major role in the deficient dendritic outgrowth. The study of 180-day-old hypothyroid rats revealed that although ectopic granule cells remained quite numerous, their number per unit surface was lesser than in the 30-day-old hypothyroid group. This finding may be related to the capacity displayed by heterotopic neurons to establish synaptic contacts with the components of the molecular layer. This was inferred by the presence of a peculiar synaptic cell investment formed by axosomatic and somatodendritic contacts in 180-day-old hypothyroid rats which shows that the surviving ectopic granule cells manage to adapt to an adverse milieu.     

Intra- and interpopulation heterogeneity of the cerebellar nerve cells in topography and ribosomal gene content

Using the method of in situ hybridization of ribosomal DNA with DNA of isolated nuclei, a study was made of localization and relative content of ribosomal genes in the Purkinje cells and other cells of the rat cerebellum. It has been shown that various cells of cerebellum have, on the average, the same number of ribosomal genes. A subpopulation with amplified ribosomal genes have been found among the nuclei of Purkinje's cells.     

Plastic synaptic reorganization in the sensorimotor cortex of adult cats after lesioning of the contralateral cerebellar nucleus interpositus

Intact cats and animals undergoing lesioning of the contralateral cerebellar nucleus interpositus between one and six months previously were used in this research, employing intracellular recording techniques and investigating the response of corticospinal neurons (CSN) to stimulating the aforementioned nucleus, the ipsilateral cerebellar nucleus interpositus, and the ventrolateral thalamic nucleus. A reduction was found in the stage of rise to peak in monosynaptic thalamocortical EPSP in CSN of operated animals, with a low axonal conduction velocity, pointing to distant terminal dendrosomatic sprouting and formation of new synapses at proximal sections of the CSN somatodendritic membrane. Findings are presented on formation of ipsilateral interpositothalamocortical projections duplicating similar contralateral projections in intact animals. Contralateral cortico-interposital collaterals were found in intact animals and similar sprouting of ipsilateral origin in those which had undergone surgery.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 22, No. 6, pp. 761–771, November–December, 1990.     

Calcium requirement of long-term depression and rebound potentiation in cerebellar Purkinje neurons

Cerebellar Purkinje neurons (PNs) receive two main excitatory inputs, from climbing fibers and parallel fibers, and inhibitory inputs, from GABAergic interneurons. The synapses formed by parallel fibers and by inhibitory interneurons on PNs are able to undergo long-lasting in efficacy. Thus, the excitatory parallel fiber-PN synapse undergoes long-term fibers. Synaptic inhibition can be potentiated by climbing fiber activity by a mechanism named rebound potentiation, resulting in a more powerful inhibitory effect of GABAergic interneurons. The induction of both long-term depression and rebound potentiation requires a transient elevation of the cytoplasmic calcium concentration ([Ca²⁺]_i). The [Ca²⁺]_i-transient is caused by Ca²⁺ entry through voltage-gated Ca²⁺ channels and, possibly, by release of Ca²⁺ from IP_3- and ryanodine-sensitive stores. Direct Ca²⁺ entry through synaptic AMPA receptor channels seems not to contribute significantly to the Ca²⁺ signal mediating the induction of both long-term depression and rebound potentiation.     

Linear systems analysis of the relationship between firing of deep cerebellar neurons and the classically conditioned nictitating membrane response in rabbits

The correlation of the activity of neurons in the interposed and dentate nuclei of the cerebellum with conditioned movements of the nictitating membrane was investigated using linear systems analysis. The activity of single deep cerebellar nuclear cells was assumed to be the input to a linear system that produced nictitating membrane movement. Data were initially analyzed with a causal model to assess the degree to which past neural activity predicted the conditioned response. 55 of 165 cells had correlation coefficients of 0.50 or greater between the model's moment-to-moment output and the actual output, with two interpositus cells having correlation coefficients of greater than 0.90. Double-sided impulse responses indicated that afference from the face and efference copy probably affect deep cerebellar neural activity. Nonlinearities were also found in the relationship between neuronal activity and conditioned movement. It was concluded that cerebellar deep nuclear firing is highly correlated with future nictitating membrane movements but that the firing-movement relationship contains noncausal and nonlinear components.     

Mathematical models of cochlear nucleus onset neurons: I. Point neuron with many weak synaptic inputs

The cochlear nucleus (CN) presents a unique opportunity for quantitatively studying input-output transformations by neurons because it gives rise to a variety of different response types from a relatively homogeneous input source, the auditory nerve (AN). Particularly interesting among CN neurons are Onset (On) neurons, which have a prominent response to the onset of sustained sounds followed by little or no response in the steady-state. On neurons contrast sharply with their AN inputs, which respond vigorously throughout stimuli. On neurons can entrain to stimuli (firing once per cycle of a periodic stimulus) at up to 1000 Hz, unlike their AN inputs. To understand the mechanisms underlying these response patterns, we tested whether an integrate-to-threshold point-neuron model with a fixed refractory period can account for On discharge patterns for tones, systematically examining the effect of membrane time constant and the number and strength of the exclusively excitatory AN synaptic inputs. To produce both onset responses to high-frequency tone bursts and entrainment to a broad range of low-frequency tones, the model must have a short time constant (0.125 ms) and a large number (>100) of weak synaptic inputs, properties that are consistent with the electrical properties and anatomy of On-responding cells. With these parameters, the model acts like a coincidence detector with a threshold-like relationship between the instantaneous discharge rates of the output and the inputs. Onset responses to high-frequency tone bursts result because the threshold effect enhances the initial response of the AN inputs and suppresses their relatively lower sustained response. However, when the model entrains across a broad range of frequencies, it also produces short interspike intervals at the onset of high-frequency tone bursts, a response pattern not found in all types of On neurons. These results show a tradeoff, that may be a general property of many neurons, between following rapid stimulus fluctuations and responding without short interspike intervals at the onset of sustained stimuli.     

Ultrastructure of synaptic terminals in nucleus infundibularis and nucleus supraopticus of Passer domesticus

Summary Axo-dendritic and axo-somatic synapses and complex axo-dendritic junctions from the nucleus infundibularis and nucleus supraopticus of Passer domesticus were studied in the electron microscope. A morphological asymmetry, represented by clustering of synaptic vesicles (of about 500 Å diameter) at the presynaptic membrane and a thickening of and subsynaptic web formation at the postsynaptic membrane, was observed in the axo-dendritic and axo-somatic synapses, and to a smaller extent in the complex axo-dendritic junctions. In the hypothalamic synaptic axon terminals, as in the perikarya of nucleus infundibularis, dense-core vesicles of 800–1,000 Å mean diameter were observed in addition to the synaptic vesicles, but the 800–1,000 Å vesicles were not concentrated at the presynaptic membrane. Granules of 2,000 Å mean diameter, observed in the neurons of nucleus supraopticus, were not seen in any hypothalamic synaptic terminals. Intersynaptic filaments and an intrasynaptic band lying in the intersynaptic cleft were frequently observed. The significance of the synaptic structures was discussed in relation to neurotransmission and to possible mechanisms by which perikarya of the nucleus infundibularis and nucleus supraopticus receive nervous information.

---

Zusammenfassung Mit dem Elektronenmikroskop wurden im Nucleus infundibularis und Nucleus supraopticus von Passer domesticus axo-dendritische und axo-somatische Synapsen sowie komplexe axo-dendritische Kontaktstellen untersucht. Vor allem zeichnen sich die axodendritischen und axo-somatischen Synapsen durch eine Polarität ihrer Feinstruktur aus. Diese wird in erster Linie durch eine Anhäufung von etwa 500 Å großen synaptischen Bläschen an der präsynaptischen Membran sowie durch eine Verdichtung und ein subsynaptisches Geflechtwerk an der postsynaptischen Membran gekennzeichnet. Die hypothalamischen synaptischen Axonendigungen enthalten neben den charakteristischen synaptischen Bläschen elektronendichte Granula mit einem Durchmesser von etwa 800–1000 Å; die letzteren, die u.a. auch für Perikaryen des Nucleus infundibularis charakteristisch sind, lassen sich jedoch nicht in unmittelbarer Nähe der präsynaptischen Membran nachweisen. Granula mit einem Durchmesser von etwa 2000 Å, die in den Neuronen des Nucleus supraopticus zu beobachten sind, fehlen in den hypothalamischen synaptischen Nervenendigungen. Intersynaptische Filamente und ein den Synapsenspalt füllendes intrasynaptisches Band konnten in zahlreichen Synapsen dargestellt werden. Die Bedeutung der Synapsenstrukturen wird im Hinblick auf die nervöse Übertragung und auf die Mechanismen, die der Zuleitung nervöser Informationen an den Nucleus infundibularis und Nucleus supraopticus dienen, diskutiert.

---

---

Dedicated to Professor Ernst Horstmann.

---

The investigation was supported by a scholarship of the Alexander von Humboldt-Stiftung to Dr. J. Priedkalns (on leave from the Department of Veterinary Anatomy, University of Minnesota, St. Paul, U. S. A.) and by a research grant of the Deutsche Forschungsgemeinschaft to Professor A. Oksche.     

Association of tyrosine hydroxylase with synaptic vesicles in bovine caudate nucleus

Abstract— The subcellular distribution of tyrosine hydroxylase in the bovine caudate nucleus was investigated, with five other enzymes serving as biochemical markers, and coordinated with electron microscopic observations. Tyrosine hydroxylase is particulate bound and has relatively high specific activities in the synaptosome-enriched subfractions and in the synaptic vesicle fraction. The association of tyrosine hydroxylase with the synaptic vesicles is consistent with the theory of feedback regulation of catecholamine synthesis.