Neurophysiological modeling of Voiding in Rats

Like man, most animals need a regular supply of nutriments. Eating and drinking cater for this need, but not with 100% efficiency. A considerable part of the conswned goods are expelled from the body as defecation. Waste products from the blood are filtered by the kidneys and excreted as urine. 1ms process continues throughout the day. In many situations, however, it would be very inconvenient to be expelling urine. Animals that leak urine continuously are more easily traced by preditors than the ones that don't. Besides this, for mankind it is socially intolerable at certain moments. Nature supplied q solution that enables storage of urine until a convenient moment for voiding: the bladder. Upon desire the bladder contracts and urine is expelled through the urethra, which, with the bladder, constitutes ti,e lower urinary tract. Thus the bladder owner controls when and where to expel urine

Changes within bursts during learning in dissociated neural networks

We have studied the effect of imprinting a new stimulus-response (SR) relationship into a neuronal network cultured on a multi electrode array (MEA). We have used the Conditional Repetitive Stimulation (CRS) algorithm introduced by Shahaf et al in 2004. In this algorithm focal electrical stimulation is delivered at a low rate (≪1 Hz) and is withdrawn when a desired response is observed. We confirmed that CRS could train the network to strengthen an initially weak SR relationship. With the acquisition of a new SR relationship, we studied its effect on network activity. Specifically, spontaneously occurring network bursts measured before, during and after training were analyzed. The total firing rate within bursts was estimated with a temporal resolution of milliseconds (burst profiles). We have shown earlier that these profiles change shape on a time base of several hours during spontaneous development. We show that the rate of change of the profiles during training (i.e. CRS) was higher than when no stimulation was applied

Orexin A in cortical cultures: expression and effect on synaptogenesis during development

Orexin-A (OXA) is an excitatory hypothalamic neurotransmitter and ligand for Orexin Receptor-1 (OR1), isolated from a small group of hypothalamic neurons. OXA orchestrates different brain functions, and at the cognitive level some of the effects of insufficiency of OXA are well-known, for example in Parkinson’s disease. It is widely assumed that deteriorated cognitive processes are related to impaired network connectivity. However, little is known about the effects of OXA in network connectivity and synaptogenesis. Therefore, to obtain insight into this problem we designed experiments with two groups of networks of dissociated cortical neurons: one group incubated in a plain medium and another chronically treated with OXA. After one, two, three or four weeks in vitro we applied immunocytochemistry for detection of OXA, OR1 and synaptic marker synaptophysin. Shortly after plating, 91±8% of the neurons cultivated in a plain medium expressed OXA-immunoreactivity, which does normally not occur in vivo indicating that neurons may change their phenotype under non-natural culture conditions to develop synaptically coupled networks. The fraction of orexinergic neurons decreased to 33±21% after 4 weeks in vitro. OXA expression was highest in the first week of network formation, the period of maximum synaptogenesis, and then decreased and stabilized in the weeks thereafter. Our hypothesis that OXA plays a role in the network development as a synaptogenic factor was supported by higher levels, earlier onset, and sustained increase of synaptophysin-expression in experiments with chronic OXA application to the culture medium

Analysis of Cultured Neuronal Networks Using Intraburst Firing Characteristics

It is an open question whether neuronal networks, cultured on multielectrode arrays, retain any capability to usefully process information (learning and memory). A necessary prerequisite for learning is that stimulation can induce lasting changes in the network. To observe these changes, one needs a method to describe the network in sufficient detail, while stable in normal circumstances. We analyzed the spontaneous bursting activity that is encountered in dissociated cultures of rat neocortical cells. Burst profiles (BPs) were made by estimating the instantaneous array-wide firing frequency. The shape of the BPs was found to be stable on a time scale of hours. Spatiotemporal detail is provided by analyzing the instantaneous firing frequency per electrode. The resulting phase profiles (PPs) were estimated by aligning BPs to their peak spiking rate over a period of 15 min. The PPs reveal a stable spatiotemporal pattern of activity during bursts over a period of several hours, making them useful for plasticity and learning studies. We also show that PPs can be used to estimate conditional firing probabilities. Doing so, yields an approach in which network bursting behavior and functional connectivity can be studied

Pudendal nerve stimulation induces urethral contraction and relaxation

In this study we measured urethral pressure changes in response to efferent pudendal nerve stimulation in rats. All other neural pathways to the urethra were transected, and the urethra was continuously perfused. We found fast twitch-like contractions, superimposed on a slow relaxation. The amplitude of the twitches was independent of the stimulation frequency below 26 Hz, whereas the relaxation depended highly on this frequency. The twitches were caused by striated urethral muscles, and the relaxation was caused by smooth muscles. Both were mediated by acetylcholine. We calculated the effective urethral relaxation as the absolute relaxation multiplied by the time fraction between the twitches. Maximum effective relaxation occurred at 8-10 Hz, exactly the frequency of spontaneous oscillations during bladder voiding in rats. Although the oscillatory sphincter contractions in rats during voiding may be needed in other mechanisms for efficient voiding, our data suggest that they may be a side effect of the actual purpose: urethral relaxation

The Effect of Slow Electrical Stimuli to Achieve Learning in Cultured Networks of Rat Cortical Neurons

Learning, or more generally, plasticity may be studied using cultured networks of rat cortical neurons on multi electrode arrays. Several protocols have been proposed to affect connectivity in such networks. One of these protocols, proposed by Shahaf and Marom, aimed to train the input-output relationship of a selected connection in a network using slow electrical stimuli. Although the results were quite promising, the experiments appeared difficult to repeat and the training protocol did not serve as a basis for wider investigation yet. Here, we repeated their protocol, and compared our ‘learning curves’ to the original results. Although in some experiments the protocol did not seem to work, we found that on average, the protocol showed a significantly improved stimulus response indeed. Furthermore, the protocol always induced functional connectivity changes that were much larger than changes that occurred after a comparable period of random or no stimulation. Finally, our data shows that stimulation at a fixed electrode induces functional connectivity changes of similar magnitude as stimulation through randomly varied sites; both larger than spontaneous connectivity fluctuations. We concluded that slow electrical stimulation always induced functional connectivity changes, although uncontrolled. The magnitude of change increased when we applied the adaptive (closed-loop) training protocol. We hypothesize that networks develop an equilibrium between connectivity and activity. Induced connectivity changes depend on the combination of applied stimulus and initial connectivity. Plain stimuli may drive networks to the nearest equilibrium that accommodates this input, whereas adaptive stimulation may direct the space for exploration and force networks to a new balance, at a larger distance from the initial state

Neurophysiological modeling of voiding in rats: urethral nerve response to urethral pressure and flow

In male urethan-anesthetized rats, activity was measured in nerves that run over the proximal urethra. The urethral nerve response to stepwise urethral perfusion could be described by a four-parameter model (fit error < 6%). At the onset of perfusion, the urethra was closed and the pressure increased with the infused volume. The nerve activity (NA) increased linearly with this inserted volume to a maximum (NAmax), which was proportional to the instantaneous pressure. The duration of this first episode (delta t) was inversely proportional to the perfusion rate. After infusion of a fixed volume, the urethra opened and the NA decreased with a time constant phi -1 (approximately 1.8 s) to an elevated level (NAlevel). NAlevel was linearly related to the steady-state pressure. Accordingly, sensors in the urethra are sensitive to pressure rather than to the perfusion rate. The parameters NAmax, NAlevel, and delta t showed very good reproducibility (SD approximately 19% of mean). The measured activity was most likely afferent and conducted to the major pelvic ganglion

Threshold for efferent bladder nerve firing in the rat

In this study, the mechanism involved in the initiation of voiding was investigated. Bladder pressure and bladder and urethral nerve activity were recorded in the anesthetized rat. Bladder nerve activity was resolved into afferent and efferent activity by means of a theoretical model. The beginning of an active bladder contraction was defined as the onset of bladder efferent firing at a certain time (t0). From t0 onward, bladder efferent activity increased linearly during deltat seconds (rise time) to a maximum. The pressure at t0 was 1.0 +/- 0.4 kPa, the afferent nerve activity at t0 was 2.0 +/- 0.6 microV (53 +/- 15% of maximum total nerve activity), and deltat was 11 +/- 13 s. Between contractions the afferent activity at t0 was never exceeded. Urethral afferent nerve activity started at bladder pressures of 2.1 +/- 1.1 kPa. Therefore, we concluded that urethral afferent nerve activity does not play a role in the initiation of bladder contractions; voiding contractions presumably are initiated by bladder afferent nerve activity exceeding a certain threshold

Prediction in cultured cortical neural networks

Theory suggest that networks of neurons may predict their input. Prediction may underlie most aspects of information processing and is believed to be involved in motor and cognitive control and decision-making. Retinal cells have been shown to be capable of predicting visual stimuli, and there is some evidence for prediction of input in the visual cortex and hippocampus. However, there is no proof that the ability to predict is a generic feature of neural networks. We investigated whether random in vitro neuronal networks can predict stimulation, and how prediction is related to short- and long-term memory. To answer these questions, we applied two different stimulation modalities. Focal electrical stimulation has been shown to induce long-term memory traces, whereas global optogenetic stimulation did not. We used mutual information to quantify how much activity recorded from these networks reduces the uncertainty of upcoming stimuli (prediction) or recent past stimuli (short-term memory). Cortical neural networks did predict future stimuli, with the majority of all predictive information provided by the immediate network response to the stimulus. Interestingly, prediction strongly depended on short-term memory of recent sensory inputs during focal as well as global stimulation. However, prediction required less short-term memory during focal stimulation. Furthermore, the dependency on short-term memory decreased during 20 h of focal stimulation, when long-term connectivity changes were induced. These changes are fundamental for long-term memory formation, suggesting that besides short-term memory the formation of long-term memory traces may play a role in efficient prediction.</p