AGING-ASSOCIATED COGNITIVE DECLINE IS REVERSED BY D-SERINE SUPPLEMENTATION.

Brain aging is a natural process that involves structural and functional changes that lead to cognitive decline, even in healthy subjects. This detriment has been associated with N-methyl-D-aspartate receptor (NMDAR) hypofunction due to a reduction in the brain levels of D-serine, the endogenous NMDAR co-agonist. However, it is not clear if D-serine supplementation could be used as an intervention to reduce or reverse age-related brain alterations. In the present work, we aimed to analyze the D-serine effect on aging-associated alterations in cellular and large-scale brain systems that could support cognitive flexibility in rats. We found that D-serine supplementation reverts the age-related decline in cognitive flexibility, frontal dendritic spine density, and partially restored large-scale functional connectivity without inducing nephrotoxicity; instead, D-serine restored the thickness of the renal epithelial cells that were affected by age. Our results suggest that D-serine could be used as a therapeutic target to reverse age-related brain alterations.SIGNIFICANT STATEMENTAge-related behavioral changes in cognitive performance occur as a physiological process of aging. Then, it is important to explore possible therapeutics to decrease, retard or reverse aging effects on the brain. NMDA receptor hypofunction contributes to the aging-associated cognitive decline. In the aged brain, there is a reduction in the brain levels of the NMDAR co-agonist, D-Serine. However, it is unclear if chronic D-serine supplementation could revert the age-detriment in brain functions. Our results show that D-serine supplementation reverts the age-associated decrease in cognitive flexibility, functional brain connectivity, and neuronal morphology. Our findings raise the possibility that restoring the brain levels of D-serine could be used as a therapeutic target to recover brain alterations associated with aging.

Axons of Individual Dorsal Horn Neurons Bifurcated to Project in Both the Anterolateral and the Postsynaptic Dorsal Column Systems.

Sensory information stimulates receptors of somatosensory system neurons generating a signal that codifies the characteristics of peripheral stimulation. This information reaches the spinal cord and is relayed to supra-spinal structures through two main systems: the postsynaptic dorsal column-medial lemniscal (DC-ML) and the anterolateral (AL) systems. From the classical point of view, the DC-ML has an ipsilateral ascending pathway to the Gracilis (GRA) or Cuneate (CUN) nuclei and the AL has a contralateral ascending pathway to the ventral posterolateral (VPL) thalamic nucleus. These two systems have been the subject of multiple studies that established their independence and interactions. To analyze the ascending projections of L1-L5 spinal dorsal horn neurons in the rat, two retrograde neuronal tracers were injected into the GRA and the VPL. Additionally, an electrophysiological study was performed by applying electrical stimulation at the GRA or VPL and recording antidromic evoked activity in single unit spinal cord cells. Importantly, a subset of spinal dorsal horn neurons exhibited double staining, indicating that these neurons projected to both the GRA and the VPL. These double-stained neurons were located on both sides of the dorsal horn of the spinal cord. The spinal dorsal horn neurons exhibited antidromic and collision activities in response to both GRA and VPL electrical activation. These results show spinal cord neurons with bifurcated bilateral projections to both the DC-ML and AL systems. Based on these results, we named these neurons bilateral and bifurcated cells.

Prolactin fractions from lactating rats elicit effects upon sensory spinal cord cells of male rats.

Recently it has been suggested that the neurohormone prolactin (PRL) could act on the afferent nociceptive neurons. Indeed, PRL sensitizes transient receptor potential vanilloid 1 (TRPV1) channels present in nociceptive C-fibers and consequently reduces the pain threshold in a model of inflammatory pain. Accordingly, high plasma PRL levels in non-lactating females have been associated with several painful conditions (e.g. migraine). Paradoxically, an increase of PRL secretion during lactation induced a reduction in pain sensitivity. This difference could be attributed to the fact that PRL secreted from the adenopituitary (AP) is transformed into several molecular variants by the suckling stimulation. In order to test this hypothesis, the present study set out to investigate whether PRL from AP of suckled (S) or non-suckled (NS) lactating rats affects the activity of the male Wistar wide dynamic range (WDR) neurons. The WDR neurons are located in the dorsal horn of the spinal cord and receive input from the first-order neurons (Ab-, Ad- and C-fibers). Spinal administration of prolactin variant from NS rats (NS-PRL) or prolactin variant from S rats (S-PRL) had no effect on the neuronal activity of non-nociceptive Ab-fibers. However, the activities of nociceptive Ad-fibers and C-fibers were: (i) increased by NS-PRL and (ii) diminished by S-PRL. Either NS-PRL or S-PRL enhanced the post-discharge activity. Taken together, these results suggest that PRL from S or NS lactating rats could either facilitate or depress the nociceptive responses of spinal dorsal horn cells, depending on the physiological state of the rats.

Spinal LTP induced by sciatic nerve electrical stimulation enhances posterior triangular thalamic nociceptive responses.

Long-term potentiation (LTP) can be induced by electrical stimulation and gives rise to an increase in synaptic strength at the first relay. This phenomenon has been associated with learning and memory and also could be the origin of several pathological states elicited by an initial strong painful stimulus, such as some forms of neuropathic pain. We used high-frequency electrical stimulation of the sciatic nerve in anesthetized rats to produce spinal LTP. To evaluate the effect of spinal LTP on the activity of neurons in the posterior triangular nucleus of the thalamus (PoT), we applied an electrical stimulation (40 stimuli; 1ms; 0.5Hz; 1.5mA) to cutaneous tissues at 10-min intervals during at least 3h. In the majority of cases, PoT cells did not respond to cutaneous stimulation before LTP, but 50min after LTP induction PoT cells progressively began responding to the cutaneous stimulation. Furthermore, after 3h of LTP induction, PoT neurons could respond to cutaneous stimulation applied to different paws. Interestingly, the conduction velocities for the receptive field responses from the paw to the PoT cells were compatible with those of Aδ-fibers. Since PoT cells project to the insular cortex, the progressive increase in PoT activity and also the progressive unmasking of somatic receptive fields in response to LTP, place these cells in a key position to detect pain stimuli following central sensitization.

Identification of oxytocin receptor in the dorsal horn and nociceptive dorsal root ganglion neurons.

Oxytocin (OT) secreted by the hypothalamo-spinal projection exerts antinociceptive effects in the dorsal horn. Electrophysiological evidence indicates that OT could exert these effects by activating OT receptors (OTR) directly on dorsal horn neurons and/or primary nociceptive afferents in the dorsal root ganglia (DRG). However, little is known about the identity of the dorsal horn and DRG neurons that express the OTR. In the dorsal horn, we found that the OTR is expressed principally in neurons cell bodies. However, neither spino-thalamic dorsal horn neurons projecting to the contralateral thalamic ventral posterolateral nucleus (VPL) and posterior nuclear group (Po) nor GABaergic dorsal horn neurons express the OTR. The OTR is not expressed in skin nociceptive terminals or in dorsal horn nociceptive fibers. In the DRG, however, the OTR is expressed predominantly in non-peptidergic C-fiber cell bodies, but not in peptidergic or mechanoreceptor afferents or in skin nociceptive terminals. Our results suggest that the antinociceptive effects of OT are mediated by direct activation of dorsal horn neurons and peripheral actions on nociceptive, non-peptidergic C-afferents in the DRG.

Functional interactions between the paraventricular hypothalamic nucleus and raphe magnus. A comparative study of an integrated homeostatic analgesic mechanism.

This work compares the effects of electrical stimulation of the paraventricular hypothalamic nucleus (PVN) and the raphe magnus nucleus (RMg) on the single-unit response from dorsal spinal cord neurons activated by nociceptive receptive field stimulation. We evaluated the effects of stimulating the PVN or RMg individually or simultaneously, as well as PVN stimulation after RMg electrolytic lesion. PVN or RMg stimulation suppressed the A-delta, C fiber, and postdischarge, and we demonstrated that their simultaneous stimulation increases the duration and intensity of suppressive effects. RMg lesion increased the peripheral responses, but PVN stimulation continued to be suppressive. The intrathecal administration of 20 µl of a 10⁻5 M solution of a specific oxytocin antagonist strongly reduced the PVN effects, and 20 µl of 10⁻6 M naloxone significantly reduced the RMg suppression of receptive field responses. Some spinal cord cells presented a short-latency, evoked action potential (6.8 ms and a variability of ±0.5 ms) produced by the RMg stimulation. This is interpreted as a direct postsynaptic action of the RMg on the spinal cord cells. We never found similar responses produced by the PVN, and therefore, we propose that the PVN effects are presynaptic. Finally, the immunohistochemical experiments confirmed the oxytocinergic and the vasopresinergic innervation used by the PVN projection to the RMg, and they raise the possibility that other neurotransmitters are involved. We conclude that the PVN and the RMg form part of a homeostatic analgesic mechanism acting on the same spinal cord cells to block the noxious information, but using different mechanisms. Both structures, and others, contribute to the homeostatic mechanism of endogenous analgesia.

Amplitude of somatosensory cortical evoked potentials is correlated with spontaneous activity of spinal neurones in the cat.

Simultaneous recordings of cortical evoked potentials in the posterior sigmoid gyrus, and spontaneous negative cord dorsum potentials (CDPs) of the L6 lumbar spinal segment, were made in the anaesthetised cat. The electrodes were positioned in cortical and spinal somatosensory regions where the largest spontaneous and evoked negative potentials were detected. Evoked potentials were produced by electrical stimulation to cutaneous nerves or by mechanical stimulation of the hindpaw skin. We found that both electrically and mechanically cortical evoked potentials were facilitated during the spontaneous negative CDPs. The magnitude of such facilitation was proportional to the amplitude of the 'conditioning' spontaneous negative CDPs. This led to a high positive correlation between amplitude fluctuations of spontaneous negative CDPs and fluctuations of the cortical evoked potentials. This observation suggests that transmission of cutaneous sensory information in ascending pathways could be facilitated when dorsal horn spinal neurones are active.

Cortical neuronal ensembles driven by dorsal horn spinal neurones with spontaneous activity in the cat.

Simultaneous recordings of cortical activity, recorded as the cortical local field potential (CLFP) in the contralateral posterior sigmoid gyrus, and the spinal activity, recorded as the cord dorsum potential (CDP) of the L6 lumbar segment, were made in the anaesthetized cat. The electrodes were positioned in somatosensory regions where the largest spontaneous negative CLFPs and CDPs were recorded. We found that spontaneous negative CLFPs were preceded by spontaneous negative CDPs with a mean latency of 14.4+/-3.5 ms. Amplitude of these spontaneous negative CLFPs was abolished after section of the dorsal columns and ipsilateral dorsolateral funiculus. It is concluded that the neurones of the primary somatosensory cortex can be driven by dorsal horn spinal neurones producing the spontaneous negative CDPs. This suggests very strongly that spontaneous neuronal activity in somatosensory regions of the brain is generated not only by ongoing activity of neurones located at supraspinal sites, but also by ongoing activity of spinal neurones.

Nitric oxide modulates spontaneous cord dorsum potentials in the cat spinal cord.

A previous study has shown that lumbar spontaneous cord dorsum potentials (CDPs) are produced by background activity of a neuronal ensemble located in the dorsal horn. Here, the effects produced by intravenous application of the nitric oxide synthase inhibitor L-N(G)-nitro arginine (L-NOARG, 100 microg/kg) and of the nitric oxide donor 3-morpholinosydnonimine hydrochloride (SIN-1, 500 microg/kg) on spontaneous CDPs were examined. Experiments were performed on pentobarbitally anesthetized, paralyzed and spinalized cats. The amplitude of spontaneous CDPs increased after L-NOARG, however, decreased after SIN-1. These observations suggest that electrical activity of dorsal horn neurones generating spontaneous CDPs is dependent on nitric oxide production.

NO donor SIN-1 potentiates monosynaptic reflexes in the cat spinal cord.

The effect produced by the nitric oxide donor SIN-1 on monosynaptic reflexes was examined. Experiments were performed on anesthetized, paralyzed and spinalized cats. Lumbar monosynaptic reflexes were produced by stimulation of Ia afferents. I.v. application of SIN-1 (500 microg/kg) produced a mean marked potentiation of 704% of pre-drug control (100%) in the amplitude of monosynaptic reflexes. In addition, in other experiments a concentration-dependent effect on the amplitude of monosynaptic reflexes was observed after microinjections of SIN-1 into the ventral horn (1 microl; 10(-12) - 10(-3) M), with a mean facilitatory effect of 355%. In both cases, the potentiation was reversible 45 min after i.v. or local application of SIN-1. These results provide the first evidence that monosynaptic reflexes can be potentiated by nitric oxide.

La actividad de las células excitadoras e inhibidoras de la médula rostroventromedial está modulada por la estimulación vaginocervical / The acti vity of ON and OFF cells at the rostro ventromedial medulla is modulated by vagino-cervical stimulation

En ratas anestesiadas se estudió si la actividad de las células excitadoras e inhibidoras de la médula ro s t ro v e n t romedial estaba modulada por la estimulación mecánica del cuello uterino. Las células excitadoras se identificaron por un brusco aumento de la frecuencia de impulsos antes de p roducirse el movimiento de la cola en respuesta a un estímulo térmico doloroso. Las células inhibidoras se identificaron por un súbito descenso de la frecuencia de impulsos justo antes de producirse el movimiento de la cola. Todas las células excitadoras identificadas (27 de 27) re d u j e ron su f recuencia de impulsos inmediatamente después de la aplicación de un estímulo vaginal. El efecto del estímulo vaginal en la actividad de las células persistió durante todo el periodo de estimulación. Por otra parte, todas las células inhibidoras identificadas (19 de 19) aumentaron su frecuencia de impulsos después del estímulo vaginal. El efecto del estímulo vaginal en la actividad de estas células persistiótambién durante todo el periodo de estimulación. La actividad de las células neutrales no mostró cambio alguno, ni durante la aplicación de un estímulo térmico doloroso ni durante el estímulo vaginal. Estos resultados sugieren que el efecto analgésico producido por el estímulo vaginal puede estar mediado por la actividad del circuito antinociceptivo en la médula rostroventromedial. Además, se ha sugerido que el influjo aferente del tracto genital puede inducir la actividad del circuito antinociceptivo en la médula rostroventromedial, ya sea mediante proyecciones a la sustancia gris periacueductal o proyecciones directas a la médula rostroventromedial (AU)

The activity of ON and OFF cells at the rostroventromedial medulla is modulated by vagino-cervical stimulation.

In anesthetized rats it was tested whether or not the activity of the ON and OFF cells within the rostral ventromedial medulla (RVM) is modulated by the mechanical stimulation of the uterine cervix (VS). ON cells were identified by an abrupt increase in their firing rate before the tail flick in response to a noxious heat. OFF cells were identified by a sudden decrease in their firing rate before the tail flick. All (27 out of 27) identified ON cells decreased their firing rate immediately after VS was applied. The effect of VS on the activity of the cells persisted for the entire stimulation period. On the other hand, all (19 out of 19) identified OFF cells increased their firing rate immediately after VS. The effect of VS on the activity of these cells also persisted for the entire stimulation period. The activity of the neutral cells showed no change, neither during the application of noxious heat, nor during VS. These results suggest that the analgesic-like effect produced by VS can be mediated by the activity of the antinociceptive circuit at the RVM. Alternatively, it can be suggested that the afferent inflow from the genital tract can induce the activity of the antinociceptive circuit at RVM, either by projections to the periaqueductal gray matter or by direct projections to RVM.