Juan Javier López-Guerrero

Calcium-activated potassium currents differentially modulate respiratory rhythm generation

The pre‐Bötzinger complex (PBC) generates eupnea and sighs in normoxia and gasping during hypoxia through particular mixtures of intrinsic and synaptic properties. Among intrinsic properties, little is known about the role of Ca2+‐activated potassium channels in respiratory rhythms generation. To examine this role, we tested the effects of openers and blockers of the large‐conductance (BK) and small‐conductance (SK) Ca2+‐activated potassium channels on the respiratory rhythms recorded both in vitro and in vivo, as well as on the discharge pattern of respiratory neurons in the PBC. Activation of SK channels with 1‐ethyl‐2‐benzimidazolinone (1‐EBIO) abolished sigh‐like activity and inhibited eupneic‐like activity, whereas blockade of SK channels with apamine (APA) increased frequency in both rhythms. In hypoxia, APA did not affect the transition to gasping‐like activity. At the cellular level, activation of SK channels abolished pacemaker activity and decreased non‐pacemaker neurons discharge; opposite effects were observed with SK blockade. In contrast to SK channel modulation, either activation or blockade of BK channels with NS 1619 or iberiotoxin and paxilline, respectively, produced mild effects on eupneic‐like and sigh‐like bursts during normoxia in vitro. However, BK blockers prevented the changes associated with the transition to gasping‐like activity in vitro and perturbed gasping generation and autoresuscitation in vivo. At the cellular level BK channel modulation did not affect respiratory neurons discharge. We conclude that KCa participate in rhythm generation in a state‐dependent manner; SK channels are preferentially involved in rhythm generation in normoxia whereas BK channels participate in the transition to gasping generation in hypoxia.

Somatostatin modulates generation of inspiratory rhythms and determines asphyxia survival

Breathing and the activity of its generator (the pre-Bötzinger complex; pre-BötC) are highly regulated functions. Among neuromodulators of breathing, somatostatin (SST) is unique: it is synthesized by a subset of glutamatergic pre-BötC neurons, but acts as an inhibitory neuromodulator. Moreover, SST regulates breathing both in normoxic and in hypoxic conditions. Although it has been implicated in the neuromodulation of breathing, neither the locus of SST modulation, nor the receptor subtypes involved have been identified. In this study, we aimed to fill in these blanks by characterizing the SST-induced regulation of inspiratory rhythm generation in vitro and in vivo. We found that both endogenous and exogenous SST depress all preBötC-generated rhythms. While SST abolishes sighs, it also decreases the frequency and increases the regularity of eupnea and gasping. Pharmacological experiments showed that SST modulates inspiratory rhythm generation by activating SST receptor type-2, whose mRNA is abundantly expressed in the pre-Bötzinger complex. In vivo, blockade of SST receptor type-2 reduces gasping amplitude and consequently, it precludes auto-resuscitation after asphyxia. Based on our findings, we suggest that SST functions as an inhibitory neuromodulator released by excitatory respiratory neurons when they become overactivated in order to stabilize breathing rhythmicity in normoxic and hypoxic conditions.

Vasorelaxant Effect of Mexican Medicinal Plants on Isolated Rat Aorta

Abstract The vasorelaxant effect of methanol extracts (0.86–50 µg/ml) of Iresine calea, Psyttacanthus calyculathus. (DC.) G. Don, Laelia autumnalis. (Lex.) Lindley, Brickellia cavanillesii. (Cass.) Gray, and Lepechinia caulescens. (Ortega) Epling, plant species used in Mexican folk medicine for the treatment of hypertension and related diseases, were evaluated in isolated rat aortic rings. The extracts of I. calea. and P. calyculathus. did not show a vasorelaxant activity on norepinephrine-evoked contraction (NE, 1 × 10−7.5 M) in endothelium-intact (+ E) and endothelium-denuded (− E) rat aorta rings. On the other hand, L. autumnalis. and B. cavanillesii. induced a concentration-dependent and endothelium-independent relaxation on rat aorta. However, the methanol extract of L. caulescens. produced a significant vasodilator effect in a concentration-dependent and endothelium-dependent manner. In order to determine the mode of the vasorelaxant action evoked by L. caulescens., the extract was evaluated in the presence of L-NAME (inhibitor of nitric oxide synthase at 1 × 10−4 M) and indomethacin (inhibitor of cyclooxygenases at 1 × 10−5 M). Relaxation was blocked by L-NAME, indicating the extract vasodilating properties are endothelium mediated due to liberation of nitric oxide.

Amyloid beta 1-42 inhibits entorhinal cortex activity in the beta-gamma range: role of GSK-3.

Oscillatory activity in the entorhinal cortex has been associated with several cognitive functions. Accordingly, Alzheimer Disease-associated cognitive decline has been related to amyloid beta-induced disturbances in several of these oscillatory patterns. We have previously shown that acute application of amyloid beta inhibits the generation of slow frequency oscillations (7-20 Hz). In contrast, alterations in faster oscillations recorded in Alzheimer Disease-transgenic mice that over-express amyloid beta have been controversial. Since transgenic mice may produce complex responses due to compensatory mechanisms, we tested the effect of acute application of amyloid beta on fast oscillations (beta-gamma bursts) generated by entorhinal cortex slices in vitro in a Mg2+ -ree solution. We also explored the participation of the enzyme glycogen synthase kinase 3 (GSK-3) in this effect. Our results show that bath application of a clinically relevant concentration of amyloid beta (10 nM) activates GSK-3 and reduces the power of beta-gamma bursts in the entorhinal cortex. The reduction of beta-gamma bursts by amyloid beta is blocked by inhibiting GSK-3 either with lithium or with SB 216763. Our results suggest that amyloid beta-induced inhibition of entorhinal cortex beta-gamma activity involves GSK-3 activation, which may provide a molecular mechanism for amyloid beta-induced neural network disruption and support the use of GSK-3 inhibitors to treat Alzheimer Disease.

Inhibition of protein kinase G activity protects neonatal mouse respiratory network from hyperthermic and hypoxic stress

In spite of considerable research attention focused on clarifying the mechanisms by which the mammalian respiratory rhythm is generated, little attention has been given to examining how this neuronal circuit can be protected from heat stress. Hyperthermia has a profound effect on neuronal circuits including the circuit that generates breathing in mammals. As temperature of the brainstem increases, respiratory frequency concomitantly rises. If temperature continues to increase respiratory arrest (apnea) and death can occur. Previous research has implicated protein kinase G (PKG) activity in regulating neuronal thermosensitivity of neuronal circuits in invertebrates. Here we examine if pharmacological manipulation of PKG activity in a brainstem slice preparation could alter the thermosensitivity of the fictive neonatal mouse respiratory rhythm. We report a striking effect following alteration of PKG activity in the brainstem such that slices treated with the PKG inhibitor KT5823 recovered fictive respiratory rhythm generation significantly faster than control slices and slices treated with a PKG activator (8-Br-cGMP). Furthermore, slices treated with 8-Br-cGMP arrested fictive respiration at a significantly lower temperature than all other treatment groups. In a separate set of experiments we examined if altered PKG activity could regulate the response of slices to hypoxia by altering the protective switch to fictive gasping. Slices treated with 8-Br-cGMP did not switch to the fictive gasp-like pattern following exposure to hypoxia whereas slices treated with KT5823 did display fictive gasping. We propose that PKG activity inversely regulates the amount of stress the neonatal mammalian respiratory rhythm can endure.