Background impulse activity of neurons of the fastigial nucleus in the rat cerebellum

Background impulse activity (BIA) of neuronal elements of the fastigial nucleus (FN) of rat cerebellum were investigated for the first time with the aid of various methods of statistical analysis. A clear predominance of stationary impulse flows (IFs) was discovered, along with irregularities in such flows, variously expressed and including a significant number of realizations of a regular component of impulse activity. Nonstationary IFs constituted a special variety of BIA; they were found in 15% of the total number of neurons investigated. Serial correlation analysis of the durations of interspike intervals (ISIs), both of stationary and nonstationary IFs, allowed us to identify six main varieties of ISI dynamics in the BIA of neurons of the fastigial nucleus. The patterns of recorded IFs and variations in IFs can serve as an adequate indices of the state of cellular activity during intracellular recording from cells of the central nervous system.L. A. Orbeli Institute of Physiology, Armenian Academy of Sciences, Yerevan. Translated from Neirofiziologiya, Vol. 23, No. 4, pp. 441–450, July–August, 1991.     

Synaptic processes in red nucleus neurons produced by stimulating the entopeduncular nucleus and globus pallidus in the cat

Synaptic processes in red nucleus neurons produced by stimulating the entoped-uncular nucleus and the globus pallidus were investigated during acute experiments on cats using intracellular recording techniques. It was found that stimulating these structures produces mono- and polysynaptic excitation of rubrospinal neurons. Analysis of the time parameters of the EPSP recordings obtained suggested that they were produced by activation of the axosomatic and axodendritic synapses of rubrospinal neurons. Mechanisms of basal ganglia involvement in the integrating of red nucleus activity are examined.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 17, No. 6, pp. 809–817, November–December, 1985.     

Antidromic and synaptic potentials evoked in neurons of the lateral vestibular nucleus by stimulation of the interstitial nucleus of Cajal and Darkshevich's nucleus in cats

Effects of stimulation of the interstitial nucleus of Cajal and Darkshevich's nucleus on unit activity in the lateral vestibular nucleus of Dieters were investigated in cats anesthetized with pentobarbital. Stimulation of the above-mentioned structures was shown to lead to antidromic and orthodromic activation of Dieters neurons. Axon collateral of vestibular neurons, ascending to the above-mentioned brain-stem structures were discovered electrophysiologically. Stimulation of the nuclei of Cajal and Darkshevich was shown to evoke mono- and polysynaptic EPSPs and IPSPs in neurons of Deiters nucleus. Convergence of influences from both nuclei on the neurons studied was demonstrated. The particular features and functional role of the influences observed are discussed.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 16, No. 6, pp. 822–829, November–December, 1984.     

Synaptic processes evoked in red nucleus neurons by stimulation of the substantia nigra in cats

Synaptic processes in red nucleus neurons evoked by stimulation of different parts of the substantia nigra and nucleus interpositus of the cerebellum were investigated by an intracellular recording method in acute experiments on cats. Stimulation of this sort was shown to induce mono- and polysynaptic activation of rubrospinal neurons. Monosynaptic cerebellar and nigral excitatory influences were found to be very similar. These influences were shown to converge on the same rubrospinal neurons. The functional significance of inputs from the substantia nigra to the red nucleus for movement performance is discussed.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 13, No. 2, pp. 149–158, March–April, 1981.     

Neuronal mechanisms of interaction between the red nucleus and other brain stem structures

Axon collaterals of rubrospinal neurons running to many brain stem structures were identified in acute experiments on cats by a technique of intracellular recording of antidromic action potentials in conjunction with collision testing. A systemic principle of organization of rubrospinal influences and also a tendency toward synchronous arrival of rubrospinal impulses at various brain stem centers were demonstrated. Most of these centers are relay nuclei, sending direct afferent projections to regions of the cerebellum which, in turn, control activity of the red nucleus. Besides such a loop, effecting dynamic cerebellar control over motor function, transmission of somatosensory information from nuclei of the dorsal columns of the spinal cord directly to the red nucleus was demonstrated. Special features of mono- and polysynaptic EPSPs evoked by stimulation of nuclei of the dorsal columns indicate that such PSPs arise in different regions of the soma-dendritic membrane of red nucleus neurons. The mechanisms of integration of descending motor volleys by the red nucleus are discussed.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 16, No. 5, pp. 665–678, September–October, 1984.     

Spatial summation processes in the receptive fields of visually driven neurons of the cat's cortical area 21a

The spatial summation in receptive fields (RF) of single neurons in cat's extrastriate area 21a was investigated as a basic neurophysiological substrate for central integration processing of visual information. The results showed that the majority of investigated neurons changed their response patterns with gradual increase of applied stimulus size. In approximately 82% of cases the suppression of neuron discharges was observed when the length of the moving strip exceeded that of the RF. In some neurons the increased size of the moving stimulus leads to the changes in the RF substructure. Receptive fields of neurons recorded at the same microelectrode penetration depth showed a great variety of RF superpositions distributed in a spatially asymmetric manner. As a result, every single RF consists of multiple sub-regions within the RF, differing from each other by the number of superimposed RF-s (density factor). We suggest that such complex spatial organization of the RF provides the neurophysiological basis for central integration processing of the visual information.     

Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime

Spectral diversity of fluorescent proteins, crucial for multiparameter imaging, is based mainly on chemical diversity of their chromophores. Recently we have reported, to our knowledge, a new green fluorescent protein WasCFP—the first fluorescent protein with a tryptophan-based chromophore in the anionic state. However, only a small portion of WasCFP molecules exists in the anionic state at physiological conditions. In this study we report on an improved variant of WasCFP, named NowGFP, with the anionic form dominating at 37°C and neutral pH. It is 30% brighter than enhanced green fluorescent protein (EGFP) and exhibits a fluorescence lifetime of 5.1 ns. We demonstrated that signals of NowGFP and EGFP can be clearly distinguished by fluorescence lifetime in various models, including mammalian cells, mouse tumor xenograft, and Drosophila larvae. NowGFP thus provides an additional channel for multiparameter fluorescence lifetime imaging microscopy of green fluorescent proteins.     

Real-time differential labeling of blood, interstitium, and lymphatic and single-field analysis of vasculature dynamics in vivo

Lymph nodes are highly organized structures specialized for efficient regulation of adaptive immunity. The blood and lymphatic systems within a lymph node play essential roles by providing functionally distinct environments for lymphocyte entry and egress, respectively. Direct imaging and measurement of vascular microenvironments by intravital multiphoton microscopy provide anatomical and mechanistic insights into the essential events of lymphocyte trafficking. Lymphocytes, blood endothelial cells, and lymphatic endothelial cells express sphingosine 1-phosphate receptor 1, a key G protein-coupled receptor regulating cellular egress and a modulator of endothelial permeability. Here we report the development of a differential vascular labeling (DVL) technique in which a single intravenous injection of a fluorescent dextran, in combination with fluorescent semiconductor quantum dot particles, differentially labels multiple blood and lymphatic compartments in a manner dependent on the size of the fluorescent particle used. Thus DVL allows measurement of endothelial integrity in multiple vascular compartments and the affects or pharmacological manipulation in vascular integrity. In addition, this technique allows for real-time observation of lymphocyte trafficking across physiological barriers differentiated by DVL. Last, single-field fluid movement dynamics can be derived, allowing for the simultaneous determination of fluid flow rates in diverse blood and lymphatic compartments.     

Orientation sensitive properties of visually driven neurons in extrastriate area 21a of cat cortex

Orientation sensitive properties of extrastriate area 21a neurons were investigated. Special attention was paid to the qualitative characteristics of neuron responses to the different orientations of visual stimulus motion across neuron classical receptive fields (CRF). The results of experiments have shown that a group of neurons (31%) in area 21a with specialized responses to moving visual stimuli changed their direction selective (DS) characteristics depending on the orientation of the stimulus movement. Some neurons reveal an abrupt drop of the direction sensitivity index (DI) to certain orientation (58%), and some show significant increase of DI at one of applied orientations of stimulus motion (22%). Detailed investigation of response patterns of non-directional neurons to different orientations of stimulus motion have revealed clear-cut qualitative differences, such as different regularities in the distribution of inter-peak inhibitory intervals in the response pattern in dependence of the orientation of stimulus motion. The investigation of neuron CRF stationary functional organization did not reveal correlations between RF's spatial functional organization, and that of qualitative modulations of neuron response patterns. A suggestion was put forward, that visual information central processing of orientation discrimination is a complex integrative process that includes quantitative as well as qualitative transformations of neuron activity.