Modulation of inspiratory burst duration and frequency by bombesin in vitro.

Mammalian respiratory rhythm-generating circuits in the brainstem are subject to neuromodulation by multiple peptidergic afferent inputs controlling circuit behavior and outputs. Although functionally important, actions of neuropeptide modulators have not been fully characterized. We analyzed at cellular and circuit levels two inspiratory patterns intrinsically generated by the preBötzinger complex (preBötC) and their modulation by the neuropeptides bombesin and substance P (SP) in neonatal rat medullary slices in vitro. We found that, in recordings of hypoglossal nerve and preBötC neuron inspiratory activity, some inspiratory bursts occurring spontaneously under basal conditions have a biphasic shape with longer duration than normal inspiratory bursts and occur at a lower frequency. This biphasic burst pattern has been proposed to represent inspiratory activity underling periodic sighs. Bath-applied bombesin or SP decreased the period and increased the duration of both normal inspiratory and biphasic bursts and their underlying synaptic drives. The ratio of the biphasic long-duration burst period to the normal inspiratory burst period and the ratio of their burst durations remained the same before and after peptidergic modulation. Bombesin increased the frequency of the inspiratory rhythm in a Ca2+-independent manner and the frequency of long-duration bursts in a Ca2+-dependent manner. This finding suggests that period and burst duration coupling are due to intrinsic mechanisms controlling simultaneously timing and burst termination within the inspiratory rhythm-generating network. We propose a model in which signaling cascades activated by bombesin and SP modulate mechanisms controlling inspiratory burst frequency and duration to coordinate preBötC circuit behavioral outputs.

Substitution of extracellular Ca2+ by Sr2+ prolongs inspiratory burst in pre-Bötzinger complex inspiratory neurons.

The pre-Bötzinger complex (preBötC) underlies inspiratory rhythm generation. As a result of network interactions, preBötC neurons burst synchronously to produce rhythmic premotor inspiratory activity. Each inspiratory burst consists of action potentials (APs) on top of a 10- to 20-mV synchronous depolarization lasting 0.3-0.8 s known as inspiratory drive potential. The mechanisms underlying the initiation and termination of the inspiratory burst are unclear, and the role of Ca(2+) is a matter of intense debate. To investigate the role of extracellular Ca(2+) in inspiratory burst initiation and termination, we substituted extracellular Ca(2+) with Sr(2+). We found for the first time an ionic manipulation that significantly interferes with burst termination. In a rhythmically active slice, we current-clamped preBötC neurons (Vm ≅ -60 mV) while recording integrated hypoglossal nerve (∫XIIn) activity as motor output. Substitution of extracellular Ca(2+) with either 1.5 or 2.5 mM Sr(2+) significantly prolonged the duration of inspiratory bursts from 653.4 ± 30.7 ms in control conditions to 981.6 ± 78.5 ms in 1.5 mM Sr(2+) and 2,048.2 ± 448.5 ms in 2.5 mM Sr(2+), with a concomitant increase in decay time and area. Substitution of extracellular Ca(2+) by Sr(2+) is a well-established method to desynchronize neurotransmitter release. Our findings suggest that the increase in inspiratory burst duration is determined by a presynaptic mechanism involving desynchronization of glutamate release within the network.

Effect of repeated administration of Annona diversifolia Saff. (ilama) extracts and palmitone on rat amygdala kindling.

Annonas are consumed as fresh fruits, but, because of their effects on the central nervous system, are also used in folk medicine. The effect on rat amygdala kindling of repeated administration of Annona diversifolia hexane (100mg/kg IP or PO) and ethanol (100mg/kg, PO) leaf extracts and palmitone (10mg/kg, IP) was determined. Electrographic and/or behavioral changes were monitored during kindling-induced seizures 60minutes after treatments. Antiepileptic efficacy was evaluated with respect to afterdischarge (AD) duration, spike frequency, and/or behavioral seizure activity. Oral administration of both extracts significantly decreased spike frequency, whereas intraperitoneally administered hexane extract and palmitone only reduced AD duration. Hexane extract and palmitone exhibited anticonvulsant properties and delayed establishment of a kindling state as observed with diazepam (0.3mg/kg IP). These results reinforce the anticonvulsant properties of this plant, and palmitone and other constituents are responsible for the pharmacological effects.

Phytochemical screening and hypoglycemic activity of Carica papaya leaf in streptozotocin-induced diabetic rats

The extraction of plant constituents is essential to isolate biologically active compounds, aimed to understand their role on the treatment of diabetes. This study was designed to explore the preliminary phytochemical and physicochemical analysis of Carica papaya L., Caricaceae, leaf, and further evaluation of its hypoglycemic effect on diabetic rats. C. papaya leaves were extracted using chloroform, n-hexane or ethanol. For each extract a phytochemical screening was performed. The tests were conducted in triplicate and the qualitative and quantitative determination of the various metabolites was done using analytical standards proposed by Mexican Herbal Pharmacopoeia. The chloroform extract, containing steroids and quinones as major components, was chosen to study C. papaya biological effects. The chloroform extract was evaporated to dryness, and doses 0, 31, 62, 125 mg/kg were orally administered in 300 µl polyethylene glycol to diabetic rats; and 0 and 62 mg/kg to non-diabetic rats. After a 20-day treatment with the chloroform extract, the animals were sacrificed and blood was obtained for biochemical studies. The main effect observed was a decrease in serum glucose, triglycerides and transaminases in diabetic rats after the administration of C. papaya chloroform extract. These results confirm the potential beneficial action of C. papaya to treat the symptoms of diabetic patients.

Anticonvulsant effect of Annona diversifolia Saff. and palmitone on penicillin-induced convulsive activity. A behavioral and EEG study in rats.

PURPOSE: To evaluate hypnotic and anticonvulsant activities of Annona diversifolia Saff. and palmitone by using behavior and electroencephalographic (EEG) analysis in an experimental model of focal seizures in rats. METHODS: For hypnotic assessment, EEG analysis of polysomnographic slow-wave sleep (SWS) and rapid eye movement (REM) sleep for a 1 h period were performed after vehicle, A. diversifolia extract or palmitone, administration. For anticonvulsant effect, 60 minutes after treatments, EEG and behavior were analyzed during penicillin-induced seizures. Latency to the onset of the first paroxystic spike, first seizure and frequency, as well as seizure severity using Racine's scale, were determined. RESULTS: Palmitone, but not A. diversifolia extract, produced a delay in the latency to the SWS phase. In addition, both palmitone and extract decreased SWS duration and accumulated REM sleep phase. With regard to the seizures, both the extract and palmitone increased the latency to the onset of spikes and seizures, but also decreased the duration of penicillin-induced seizures. This reduction in the EEG recordings was associated with an attenuation in the severity of behavioral seizures. CONCLUSIONS: A. diversifolia and palmitone did not produce a sedative-hypnotic effect although both of them were effective in reducing the severity of behavioral and EEG seizures induced by penicillin in rats, suggesting that the diminution in the paroxystic activity by A. diversifolia is likely produced by palmitone through GABAergic neurotransmission. This study justifies and reinforces the traditional use of this plant in epilepsy.