Salvatore Sapienza

Role of phosphodiesterase 5 in synaptic plasticity and memory.

Phosphodiesterases (PDEs) are enzymes that break down the phosphodiesteric bond of the cyclic nucleotides, cAMP and cGMP, second messengers that regulate many biological processes. PDEs participate in the regulation of signal transduction by means of a fine regulation of cyclic nucleotides so that the response to cell stimuli is both specific and activates the correct third messengers. Several PDE inhibitors have been developed and used as therapeutic agents because they increase cyclic nucleotide levels by blocking the PDE function. In particular, sildenafil, an inhibitor of PDE5, has been mainly used in the treatment of erectile dysfunction but is now also utilized against pulmonary hypertension. This review examines the physiological role of PDE5 in synaptic plasticity and memory and the use of PDE5 inhibitors as possible therapeutic agents against disorders of the central nervous system (CNS).

Enhancement of expression of vascular endothelial growth factor after adeno-associated virus gene transfer is associated with improvement of brain ischemia injury in the gerbil.

Angiogenesis induced by growth factors may represent a rational therapy for patients with stroke. Vascular endothelial growth factor (VEGF) plays a pivotal role in angiogenesis and VEGF expression is enhanced in the post-ischemic brain. VEGF induced by brain hypoxia can lead to the growth of new vessels and may represent a natural protective mechanism improving survival after stroke. In the light of these findings we investigated changes of VEGF expression in different brain regions after intracerebroventricular injection of adeno-associated virus transferring gene for VEGF (rAAV-VEGF) in the gerbil, and after transient brain ischemic injury, we studied the effects of rAAV-VEGF injection on survival, brain edema, delayed neuronal death in the CA1 area and learning ability. Treatment with rAAV-VEGF 6 days or 12 days before ischemia significantly improves survival, brain edema and CA1 delayed neuronal death and post-ischemic learning evaluated by passive avoidance test. Animals treated with rAAV-VEGF showed in the thalamus and the cortex, a significant positive immunostaining for VEGF similar to those subjected to brain ischemia and not treated with rAAV-VEGF. These data represent a further contribution to a possible employment of gene therapy by using rAAV-VEGF in brain ischemia and indicate that thalamus and cortex may be targets for neuroprotective effects of VEGF.

Single Muscle Organization of Interposito-rubral Projections*

SummaryIn unanesthetized neuraxis intact cats microstimulation of the interpositus nucleus (IN) which activated a single flexor or extensor muscle in limbs, was used to investigate changes of unitary discharges of rubrospinal (RST) cells. Recordings were made from sites the stimulation of which excited the same muscle activated by the IN (agonist cells), its antagonist (antagonist cells) or heteronymous muscles (heteronymous cells).Cats submitted to chronic cerebellar decortication, acute brachium conjunctivum (BC) section, acute prerubral hemidecerebration or chronic prerubral hemidecerebration and contralateral BC section, were used as controls.It was shown that agonist RST cells were monosynaptically fired from IN, while antagonist cells were inhibited and the heteronymous ones were not influenced.Cerebellar efferents within the BC mediate both excitatory and inhibitory effects, but cerebellar cortex and prerubral structures were not involved in their production.

Multiple Representations of the Body and Input-Output Relationships in the Agranular and Granular Cortex of the Chronic Awake Guinea Pig

The organization of somatosensory input and the input-output relationships in regions of the agranular frontal cortex (AGr) and granular parietal cortex (Gr) were examined in the chronic awake guinea pig, using the combined technique of single-unit recording and intracortical microstimulation (ICMS). AGr, which was cytoarchitectonically subdivided into medial (AGrm) and lateral (AGrl) parts, also can be characterized on a functional basis. AGrl contains the head, forelimb, and most hindlimb representations; only a small number of hindlimb neurons are confined in AGrm. Different distributions of submodalities exist in AGr and Gr: AGr receives predominantly deep input (with the exception of the vibrissa region, which receives cutaneous input), whereas neurons of Gr respond almost exclusively to cutaneous input. The cutaneous or deep receptive field (RF) of each neuron was determined by natural peripheral stimulation. All studied neurons were activated by small RFs, with the exception of lip, nose, pinna, and limb units of lateral Gr (Grl), for which the RFs were larger. Microelectrode mapping experiments revealed the existence of three spatially separate, incomplete body maps in which somatosensory and motor representations overlap. One body map, with limbs medially and head rostrolaterally, is contained in AGr. A second map, comparable to the first somatosensory cortex (SI) of other mammals, is found in Gr, with hindlimb, trunk, forelimb, and head representations in an orderly mediolateral sequence. An unresponsive zone separates the head area from the forelimb region. A third map, with the forelimb rostrally and the hindlimb caudally, lies adjacent and lateral to the SI head area. This limb representation, which is characterized by an upright and small size compared to that found in SI, can be considered to be part of the second somatosensory cortex (SII). A distinct head representation was not recognized as properly belonging to SII, but the evidence that neurons of the SI head region respond to stimulation of large RFs located in lips, nose, and pinna leads us to hypothesize that the SII face area overlaps that of SI to some extent, or, alternatively, that the two areas strictly contiguous and the limits are ambiguous, making them difficult to distinguish. The input-output relationships were based on the results of RF mapping and ICMS in the same electrode penetration. The intrinsic specific interconnections of cortical neurons whose afferent input and motor output is related to identical body regions show a considerable degree of refinement.(ABSTRACT TRUNCATED AT 400 WORDS)

Motor cortex modulation of exteroceptive information at bulbar and thalamic lemniscal relays in the cat

We have studied the control of the primary motor cortex on the activity of lemniscal neurons in semi-chronic implanted cats. In each experiment, two to three foci in the primary motor cortex were identified by intracortical microstimulation at low threshold (up to 20 microA) for their capacity to evoke movements of contralateral single limb joints. Neurons belonging to the dorsal column nuclei (main cuneate nucleus and gracile nucleus), or to the ventral posterolateral nucleus, were sampled for their response to stimulation of the peripheral cutaneous fields, as well as the antidromic response to stimulation of the contralateral medial lemniscus and ipsilateral somatosensory cortex, respectively. These neurons were then tested for stimulation of the cortical foci using a current intensity equal to the threshold needed to evoke motor effects, although we reduced the duration of the stimulating trains; thus, we avoided evoking movements which could elicit afferent volleys along the somatosensory paths. It was found that the primary motor cortex was able to modulate the transmission of exteroceptive signals at the level of both dorsal column nuclei and ventral posterolateral nucleus with analogous modalities. In particular: (i) a high percentage of responses, with a prevalence of excitatory effects, was observed when the receptive field of the neurons topographically corresponded to, or was very close to, the joint controlled by a given cortical focus; (ii) in these cases, higher percentages of excitations were observed in tests which concerned the distal segments of limbs than the proximal segments; (iii) the percentage of responses became lower as the neuronal receptive field was located further from the cortical motor target, the pattern being more frequently inhibitory in nature. From a functional point of view, the motor cortex control appears to be organized in a very precise manner. Its excitatory nature might subserve integrative mechanisms by which exteroceptive information arising in a given limb segment would be enhanced by a motor command inducing movements of the same body part. Moreover, a better definition of the afferent input could be obtained by a simultaneous depression of neurons, which send towards the cortex signals from adjacent or more distant cutaneous regions. It can be hypothesized that such an organization of the cortical control could improve the discriminative somatosensory aspects during the execution of explorative movements, besides supplying a sharper cutaneous feedback to the motor cortex.

Motor responses evoked by microstimulation of restiform body in the cat

SummaryMotor effects produced by microstimulation of restiform body (RB) were studied in acute unanesthetized cats, using tungsten electrodes for stimulating the peduncle and bipolar steel electrodes for recording muscular activity (EMG). The main results were the following.1.Threshold microstimulation (18.24 μA±8.77 S.D.) of effective foci within RB elicited single muscle contractions of ipsilateral limbs, primarily of forelimb; overthreshold activation (32.83 μA±9.25 S.D.) of the same points produced complex movements in 61.54% of cases that involved muscles of shoulder, neck, and trunk.2.Single muscle contractions exhibited a mean latency (20.09 msec ± 2.04 S.D.) which was significantly longer than that shown by complex movements (10.00 msec±3.10 S.D.). Furthermore, a decrease in frequency of stimulating train below 300 Hz and a reduction in duration below 30 msec caused a steep rise of threshold for single muscle responses that was not observed when studying complex movements.3.Acute RB interruption between stimulating electrode and cerebellum abolished single muscle contractions; conversely, complex movements remained unmodified even when the RB was lesioned in cats chronically submitted to interruption of brachium conjunctivum (BC).4.The pathway involved in promoting RB induced single muscle activation includes interpositus nucleus, BC and rubrospinal tract. Possible modalities of RB afferent participation to the motor control are briefly discussed.

Modulatory action of noradrenergic system on spinal motoneurons in humans.

Previous findings in animals demonstrated that the noradrenergic coeruleospinal system exerts a tonic facilitation on spinal reflexes and that activation of alpha2-autoinhibitory receptors can be responsible for a disfacilitation of the spinal activity. To investigate this issue further, we examined whether this system is also involved in descending facilitatory control of spinal motoneurons in healthy humans. The H-reflex technique was utilized to assay the motoneuronal excitability. The ratio between the maximal reflex response (H) and maximal direct response (M) was determined in each subject and was calculated at 10 min intervals before and after i.v. administration of the alpha2-agonist clonidine (0.5 microg/kg). In all subjects a marked decrease of the H/M ratio, due to depression of the H response, occurred 10 min following the clonidine injection and reached its maximum within 30 min. No significant changes of blood pressure values were provoked by drug injections. These results suggest that an autoinhibitory action may be induced by alpha2-receptor activation of locus coeruleus neurons in humans, and that this device may serve as a mechanism for a myotonolytic action on spinal motoneurons.

Sychronizing and desynchronizing fastigial influences on the electrocortical activity of the cat, in acute experiments ☆

Abstract 1. 1. The EEG effects following electrical stimulation of the roof nucleus and following selective ablations of parts of it have been systematically studied in acute cat preparations. 2. 2. In addition to the well known EEG activating effect, fastigial stimulation can also bring about definite EEG synchronization. This effect is generally observed with low frequency stimuli and is not dependent on haemodynamic changes or on those changes in muscular tone that fastigial stimulation is known to bring about in noncurarized preparations. The synchronizing response always occurs bilaterally and generally involves the whole extent of the hemispheral cortex. The response appears at the beginning of stimulation or after some delay and often outlasts the stimulation period for seconds or even minutes; it can still be evoked after bilateral interruption of the superior cerebellar peduncles but it disappears after severance of the inferior ones. 3. 3. Selective bilateral electrocoagulation of localized portions of the roof nuclei are followed by different EEG changes. To some extent the effects can be related to the site of the lesion. In particular, localized lesions of the rostro-medial portions are followed by clear-cut and long-lasting EEG activation, whereas, after lesions confined to the rostro-lateral portions, the opposite changes are seen, though of shorter duration. 4. 4. Experiments on selective stimulation of rostro-medial or rostro-lateral parts of the fastigial nucleus were made possible by destroying, some days before stimulation, the part whose effects were not to be tested. Under these conditions it was observed that synchronizing responses can easily be obtained to low frequency rostro-medial stimulation after destruction of the rostro-lateral portion, whereas once the rostro-medial portion has been destroyed the only effects elicitable by rostro-lateral stimulation are desynchronizing. 5. 5. To some extent synchronizing structures might be electively distributed within the rostro-medial portions of the roof nuclei. The close functional and hodological connections between the roof nuclei and the brain-stem systems of diffuse ascending projection might account for the facts observed.

Cortical modulation of thalamo-cortical neurons relaying exteroceptive information: a microstimulation study in the guinea pig

SummaryThe nature and organization of cortical influences on somatosensory thalamic neurons were investigated in the guinea pig in order to ascertain if mechanisms subserving sensory-motor integration in the thalamus are as precise as has previously been demonstrated in the agranular frontal cortex (AGr) and granular parietal cortex (Gr). The study was carried out on 14 chronically-implanted awake animals. In each experiment one or two motor foci within AGr and Gr were identified according to the region of the movement evoked by intracortical microstimulation at the lowest threshold stimulation (usually 5–15 µA). Spontaneous activity of 182 thalamo-cortical single neurons was recorded in the nucleus ventralis thalami (VT). The neurons were also identified by their response to activation of cutaneous receptive fields (RFs) located in regions of vibrissae or limbs, and then tested for cortical stimulation with a pulse intensity equal to the threshold for evoking motor effects. During the cortico-thalamic tests, the duration of stimulating trains was reduced in order to avoid the appearance of limb or vibrissa movements which could activate somatosensory ascending pathways forwarding peripheral messages to VT. The cortical control on VT neurons appears to be organized in a very precise manner. It was seen that: 1) The influences on these neurons relaying exteroceptive signals specifically emanated from AGr and Gr areas which in turn received exteroceptive input. 2) The vibrissa units responded to stimulation of foci in either AGr or Gr but the reactivity was greater upon stimulation of Gr than AGr. The incidence of responses was very high when the vibrissa RF was overlapping or adjacent to the region of the cortically-evoked vibrissa movement. The response pattern was mostly excitatory. Responses were rarely observed when vibrissa RF lay distant from the vibrissa moved by cortical stimulation. 3) Neurons with limb RFs responded constantly to stimulation of Gr foci only when the RF was overlapping or adjacent to the region of the cortical motor target; in these two conditions the response pattern was excitatory and inhibitory, respectively. Inhibitions only concerned neurons with forelimb RFs. Responses to stimulation of AGr were rarely obtained. From a functional point of view, the excitatory nature of the cortical control on thalamo-cortical VT neurons suggests that a cortical signal inducing movement of a given body part is able to enhance the afferent transmission of somatosensory messages arising in the same body part. Concerning the control on forelimb neurons, this enhancement would be further amplified by a sort of “descending” surround inhibition which impairs transmission of messages coming by adjacent body parts.

Effect of Microstimulation of Movement-Evoking Cortical Foci on the Activity of Neurons on the Dorsal Column Nuclei

Cortical foci in which stimulation produced movement in either the forelimb or hindlimb were isolated in rats. In each experiment, two foci were selected: one for movement in the forelimb, and the other in the hindlimb. Stimulation was subsequently reduced in order to avoid eliciting a movement, and the effects of this stimulation on activity of gracile and cuneate neurons were examined. Both excitation and inhibition were observed and were found to be arranged in a somatotopic manner. Excitation was almost exclusively obtained when the receptive field (RF) of a given neuron corresponded to the body surfaces overlying the joints involved in the cortically evoked movement. A high percentage of neurons with RFs on body surfaces corresponding to, or adjacent to, the region of cortically induced movement were inhibited, while the activity of neurons with RFs distant to the site of movement was seldom modified. These results suggest that cortical influences exerted on the dorsal column nuclei (DCN) in rats are organized in a somatotopic manner.