Thesis (M. S.)--Electrical and Computer Engineering, Georgia Institute of Technology, 2006. / Dr. Robert H. Lee, Committee Member ; Dr. Kurt Wiesenfeld, Committee Member ; Dr Robert J. Butera, Committee Member.

Thesis (Ph. D.)--State University of New York at Binghamton, Department of Systems Science and Industrial Engineering (Biosystem concentration), 2005. / Includes bibliographical references.

No description available.

The subiculum is a structure that forms a bridge between the hippocampus and the entorhinal cortex (EC) in the brain, and plays a major role in the memory consolidation process. It consists of different types of pyramidal neurons. Based on their firing behavior, these excitatory neurons are classified into strong burst firing (SBF), weak burst firing (WBF) and regular firing (RF) neurons. In the first part of the work, morphological differences in the different neuronal subtypes was explored by biocytin staining after classifying the neurons based on the differences in electrophysiological properties. Detailed morphological properties of these three neuronal subtypes were analyzed using Neurolucida neuron reconstruction method. Unlike the differences in their electrophysiological properties, no difference was found in the morphometric properties of these neuronal subtypes. In the second part of the thesis, experimental results on spike- timing- dependent plasticity (STDP) at the proximal excitatory inputs on the subicular pyramidal neurons of the juvenile (P15-P19) rat are described. The STDP was studied in the WBF and RF neurons. Causal pairing of a single EPSP with a single back propagating action potential (bAP) at a time interval of 10 ms failed to induce plasticity. However, increasing the number of bAPs in such EPSP-bAP pair to three at 50 Hz (bAP burst) induced LTD in both, the RF, as well as the WBF neurons. Increasing the frequency of action potentials to 150 Hz in the bAP burst during causal pairing also induced LTD in both the neuronal subtypes. However, all other STDP related experiments were performed only with the bAP bursts consisting of 3 bAPs evoked at 50 Hz. Amplitude of the causal pairing induced LTD decreased with increasing time interval between EPSP and the bAP burst. Reversing the order of the EPSP and the bAP burst in the pair induced LTP only with a short time interval of 10 ms. This finding is in contrast to most of the reports on excitatory synapses, wherein the pre-before post (causal) pairing induced LTP and vice-versa. The results of causal and anti-causal pairing were used to plot the STDP curve for the WBF neurons. In the STDP curve observed in these synapses, LTD was observed upto a causal time interval of 30 ms, while LTP was limited to 10 ms time interval. Hence, the STDP curve was biased towards LTD. These results reaffirm the earlier observations that the relative timing of the pre- and postsynaptic activities can lead to multiple types of STDP curves. Next, the mechanism of non-Hebbian LTD was studied in both, the RF and WBF neurons. The involvement of calcium in the postsynaptic neuron in plasticity induction was studied by chelating intracellular calcium with BAPTA. The results indicate that the LTD induction in WBF neurons required postsynaptic calcium, while LTD induction in the RF neurons was independent of postsynaptic calcium. Paired pulse ratio (PPR) experiments suggested the involvement of a presynaptic mechanism in the induction of LTD in the RF neurons, and not in the WBF neurons since the PPR was unaffected by the induction protocol only in the WBF neurons. LTD induction in the WBF neurons required activity of the NMDA receptors since LTD was not observed in the presence of the NMDA receptor blocker in the WBF neurons, while it was unaffected in the RF neurons. However, the RF neurons required the activity of L-type calcium channels for plasticity induction, since LTD was affected in the presence of the L-type calcium channel blockers, although the WBF neurons did not require the L-type calcium channel activity for plasticity induction. Hence, in addition to a non-Hebbian STDP curve, a novel mechanism of LTD induction has been reported, where L-type calcium channels are involved in a synaptic plasticity that is expressed via change in the release probability. The findings on the STDP in subicular pyramidal neurons may have strong implications in the memory consolidation process owing to the central role of the subiculum and LTD in it.

The effects of anaesthetic applications of isoflurane on 82 CA₁ neurons were studied in in vitro preparations (guinea pigs) using intracellular recording techniques. Various parameters of their excitability such as membrane electrical properties, action potentials and their afterhyperpolarizing potentials as well as synaptic transmission were determined during bath perfusion of clinically relevant concentrtaions of isoflurane. Concentrations of isoflurane were detected in the bath with ¹⁹fluorine nuclear magnetic resonance techniques, and were found to range between 0.02 and 0.3 mM. No consistent effects on the membrane properties were observed. When synaptic activity was blocked by tetrodotoxin, isoflurane induced a hyperpolarization (3-5 mV) without affecting input conductance which was computed from the voltage responses to injections of hyperpolarizing current pulses and the slopes of current-voltage relations for each cell. Responses to depolarizing pulses revealed that the threshold, amplitude and duration of the evoked spikes were not greatly altered, although repetitive spike firing was suppressed in a dose-dependent manner by isoflurane. Similarly, the amplitude and duration of the long-lasting hyperpolarizations following the elicitation of multiple (4 or 5) spikes were reduced in a reversible and dose-dependent manner. Reductions in amplitude and duration of excitatory and inhibitory postsynaptic potentials evoked by electrical stimulation of stratum radiatum were observed; these effects also were strictly dependent on the dose, as well as on duration of the application. These investigations have revealed that isoflurane interferes with synaptic transmission in the hippocampal slice preparation and suggest that presynaptic actions on transmitter release, in addition to postsynaptic effects / Medicine, Faculty of / Anesthesiology, Pharmacology and Therapeutics, Department of / Graduate

L'omniprésence des horloges circadiennes à travers une vaste gamme de taxons démontre la valeur adaptative de connaître l'heure du jour. Ces horloges permettent aux organismes de synchroniser leurs processus biologiques quotidiens à des environnements externes et internes changeants. Dans mon projet de doctorat, j'ai utilisé la drosophile comme système modèle pour étudier les bases neurales de l'adaptation saisonnière de l'activité quotidienne par l'horloge. Chez la drosophile, l'horloge cérébrale régulant l'activité locomotrice, suivant un modèle bimodale, fonctionne comme un réseau multi-oscillateurs. Deux ensembles distincts de neurones contrôlent l'activité du matin et l’activité du soir quotidiennement. Les neurones contribuant à l'activité du soir sont nombreux (oscillateurs E : 6 LNds, 1 sLNv et environ 12 à 15 DN1ps dans chaque hémisphère) et très divers en termes de localisations anatomiques, de motifs de projection, de neurochimie et de modalités photoréceptives. Mon travail indique que les différents oscillateurs E possèdent également des activités fonctionnelles distinctes dans le réseau neuronal d'horloge. J’ai démontré que seulement 2 paires d'oscillateurs E (ITP + CRY +) sur environ 150 neurones d'horloge sont suffisantes pour l'activité d'anticipation du soir. La dissection génétique de divers sous-ensembles d'oscillateurs du soir indique que non seulement ces deux paires de neurones sont suffisantes pour l'activité du soir, mais également qu'elles sont fonctionnellement supérieures aux autres oscillateurs du soir. Par conséquent, une hiérarchie opérationnelle existe parmi les oscillateurs du soir dans lesquels les neurones oscillateurs ITP + CRY + (dorénavant, ITP E) occupent l'échelon le plus élevé. J’ai par ailleurs démontré que cette hiérarchie est plutôt flexible, et que les partenaires de cette relation hiérarchique changent de rôle en fonction des entrées neuropeptidergiques (à savoir, le PDF). Les comportement et les réponses calciques des divers neurones du soir suggèrent que le PDF et les signaux saisonniers agissent sur un cadre fonctionnel, dans lequel certains neurones construisent l'activité du soir en augmentant l’activité en fin de journée et que d'autres neurones y contribuent en inhibant l'activité du début d'après-midi. Mise à part le PDF, les indices saisonniers, tels que la durée du jour, l'intensité lumineuse et la température, déterminent la pondération fonctionnelle parmi les oscillateurs du soir. Les signaux saisonniers influencent différents oscillateurs pour remplir la même fonction sous différentes saisons. Les oscillateurs ITP E sont recrutés principalement dans des conditions hivernales, tandis que les oscillateurs non-ITP E contribuent davantage dans des conditions semblables à celles de l'été. Ce recrutement biaisé d'oscillateurs se produit en partie via la modulation des niveaux de PDF par des indices saisonniers.Même s'il existe de nombreux oscillateurs E dans le circuit neuronal circadien, leur pertinence fonctionnelle est définie par des stimuli externes (indices saisonniers) et internes (neuropeptides) grâce au recrutement de différents oscillateurs.En résumé, mon étude de doctorat tente de fournir une explication plausible sur la manière dont l'adaptation saisonnière de l'horloge circadienne est réalisée au niveau neuronal. Mes résultats supportent l'idée que le recrutement d'oscillateurs, contrôlé par l'environnement, facilite l'ajustement saisonnier sculpté par l'horloge circadienne multi-oscillateurs.LNd: dorsal-lateral neuronssLNv: small ventral-lateral neuronsDN1p: dorsal neurons 1 (posterior)ITP: Ion Transport PeptideCRY: CryptochromePDF: Pigment Dispersing Factor / The ubiquity of circadian clocks across a vast range of taxa signifies the adaptive value of knowing the time of the day. These clocks enable organisms to synchronize their daily biological processes to changing external and internal environments. In my PhD project, I used Drosophila as a model system to test hypotheses regarding the neural basis of the seasonal adaptation of the clock-driven daily activity pattern. In Drosophila, the brain clock regulating bimodal locomotor activity functions as a multi-oscillator network. Two distinct sets of neurons control morning and evening bouts of daily locomotor activity. Neurons contributing to the evening activity (E oscillators; 6 LNds, 1 sLNv and around 12 to 15 DN1ps in each hemisphere) are numerous and quite diverse within themselves in terms of their anatomical loci, projection patterns, neurochemistry, and photoreceptive modalities. My work indicates that the different E oscillators also possess distinct functional loci in the clock neuronal network. I show that only 2 pairs of E oscillators (ITP+ CRY+) out of around 150 clock neurons are sufficient for the evening anticipatory activity. Genetic dissection of various evening oscillator subsets further indicates that not only these two pairs of neurons are sufficient for the evening activity, but also, they are functionally superior to other evening oscillators in their contribution to the evening activity. Hence, an operational hierarchy exists among the evening oscillators in which the ITP+ CRY+ (henceforth, ITP E) oscillator neurons inhabit the highest rung. I further show that this hierarchy is rather flexible, and the partners of this hierarchical relationship switch roles depending on neuropeptidergic inputs (namely, PDF). Studying behavior and calcium responses in diverse evening neurons suggest that PDF and seasonal cues act on a functional framework of E neurons in which some build evening activity by promotion of activity in the later parts of the day and while others, by inhibiting activity in the earlier afternoon. Alongside PDF, seasonal cues such as day-length, light intensity and temperature, determine the functional weightage among evening oscillators. Specific seasonal cues recruit different oscillators to carry out the same function under different seasons. ITP E oscillators are recruited mostly by winter-like conditions whereas non- ITP E oscillators contribute more under summer-like conditions. This biased recruitment of oscillators partly occurs via modulation of the PDF levels by seasonal cues. Even though there are numerous E oscillators in the brain circadian circuit, their functional relevance is defined by external (seasonal cues) and internal (neuropeptides) environments through conditional oscillator recruitment. In summary, my PhD study attempts to provide a plausible explanation of how seasonal adaptation of the circadian clock and the behaviours that it times, is achieved at the neural level. My results support the idea that environmentally gated recruitment of oscillators facilitates seasonal adjustment of the daily activity pattern sculpted by the multi-oscillator circadian clock.LNd: dorsal-lateral neuronssLNv: small ventral-lateral neuronsDN1p: dorsal neurons 1 (posterior)ITP: Ion Transport PeptideCRY: CryptochromePDF: Pigment Dispersing Factor

The acetylcholinesterase (AChE) content of rat vagal efferent neurons was studied. Retrograde transport of horseradish peroxidase (HRP) by cut vagal axons provided a means for localizing efferent cell bodies; tissue sections were then processed for the simultaneous visualization of HRP and AChE. A dorsal vagal efferent column contained the dorsal motor nucleus of the vagus, as a primary component, and extended caudally into the upper cervical spinal cord. A ventral column contained neurons in the nucleus ambiguus and the surrounding reticular formation. Although most of the vagal efferent neurons stained with moderate to heave intensity for AChE there were some HRP-labeled cells that contained little AChE and a small percentage in which AChE was absent. In spite of the fact that AChE has been demonstrated in certain non-cholinergic neurons, it has also been found in all cholinergic neurons. Therefore, the presence of AChE has been regarded as a necessary (but not sufficient) component for identifying cholinergic neurons. The absence of AChE in a small percentage of the vagal efferent neurons indicates that some preganglionic parasympathetic fibers in the vagus nerve are not cholinergic.

Electrical excitation of the dorsal aspect of the rostral thoracic spinal cord imparts long-term therapeutic benefits to patients with angina pectoris. Such spinal cord stimulation also induces short-term suppressor effects on the intrinsic cardiac nervous system. The purpose of this study was to determine whether spinal cord stimulation (SCS) induces long-term effects on the intrinsic nervous system, particularly in the presence of myocardial ischaemia. The activity generated by right atrial neurons was recorded in 10 anesthetized dogs during basal states, during prolonged (15 min) occlusion of the left anterior descending coronary artery, and during the subsequent reperfusion phase. Neuronal activity and cardiovascular indices were also monitored when the dorsal T1-T4 segments of the spinal cord were stimulated electrically (50 Hz; 0.2 ms) at an intensity 90% of motor threshold (mean 0.32 mA) for 17 min. SCS was performed before, during and after 15-min periods of regional ventricular ischaemia. Occlusion of a major coronary artery, one that did not perfuse investigated neurons, resulted in their excitation. Ischaemia-induced neuronal excitatory effects were suppressed (-76% from baseline) by SCS. SCS suppression of intrinsic cardiac neuronal activity persisted during the subsequent reperfusion period; after terminating 17 min of SCS, at least 20 min elapsed before intrinsic cardiac neuronal activity returned to baseline values. It is concluded that populations of intrinsic cardiac neurons are activated by inputs arising from the ischaemic myocardium. Ischaemia-induced activation of these neurons is nullified by SCS. The neuronal suppressor effects that SCS induces persist not only during reperfusion, but also for an extended period of time thereafter. These long-term effects may account, in part, for the fact that SCS imparts clinical benefit to patients with angina of cardiac origin not only during its application, but also for a time thereafter.

The electrophysiological and morphological properties of small networks of mammalian neurons were investigated with mouse spinal cord monolayer cultures of 2 mm diameter grown on multimicroelectrode plates (MMEPs). Such cultures were viewed microscopically and their activity simultaneously recorded from 2 of any 36 fixed recording sites. The specific aims achieved were: development of techniques for production of functional MMEPs and maintenance of mini-cultures, characterization of the spontaneous activity of mini-cultures, application of inhibitory and disinhibitory agents, development of staining methods for cultured neurons and initial light microscopic analysis with correlation of electrophysiological and morphological characteristics.

No description available.