Treatment of clinically nonfunctioning pituitary adenomas with dopamine agonists.

OBJECTIVE: Clinically nonfunctioning pituitary adenoma (NFPA) remains the only pituitary tumor subtype for which no effective medical therapy is available or recommended. We evaluated dopamine agonist (DA) therapy for preventing growth of postsurgical pituitary tumor remnants. DESIGN: The study design included historical cohort analysis of clinical results at two pituitary referral centers with different standard practices for postoperative NFPA management: DA therapy or conservative follow-up. METHODS: Seventy-nine patients followed for 8.8±6.5 years were treated with DA, initiated upon residual tumor detection on postoperative MRI (preventive treatment (PT) group, n=55), or when tumor growth was subsequently detected during follow-up (remedial treatment (RT) group, n=24). The control group (n=60) received no medication. Tumoral dopamine and estrogen receptor expression assessed by quantitative RT-PCR and immunostaining were correlated with response to treatment. RESULTS: Tumor mass decreased, remained stable, or enlarged, respectively, in 38, 49, and 13% of patients in the PT group, and in 0, 53, and 47% of control subjects; shrinkage or stabilization was achieved in 58% of enlarging tumors in the RT group, P < 0.0001.Fifteen-year progression-free survival rate was 0.805, 0.24, and 0.04, respectively, for PT, RT, and control groups (P<0.001). About 42% of patients in the control group required additional surgery or radiotherapy, compared with 38 and 13% subjects in the RT and PT groups, respectively (P=0.002). Outcome measures were not related to NFPA D2R abundance. CONCLUSIONS: Dopamine agonist therapy in patients with NFPA is associated with decreased prevalence of residual tumor enlargement after transsphenoidal surgical resection.

Dynamic and spatial features of the inhibitory pallidal GABAergic synapses.

The globus pallidus, one of the basal ganglia nuclei, plays a major role in both basal ganglia physiology and pathophysiology. The globus pallidus is innervated mainly by striatal spiny neurons and globus pallidus collaterals. These GABAergic synapses constitute 90% of the input to globus pallidus cells. Despite the dominance of this inhibitory GABAergic input, globus pallidus cells are spontaneously active and most of them increase their firing rate in a task related manner. To explain this apparent inconsistency, we studied the dynamic and spatial effects of GABAergic inputs to globus pallidus neurons. To this end, we used intra-cellular recording from globus pallidus neurons in rat brain slices, investigating the effect of bath and local GABA application, as well as the responses to electrical stimulation of the striatum. We showed that the properties of the responses to either local or global GABA applications are similar to the responses of globus pallidus cells to GABA release from nerve terminals. Since the stimulus-evoked responses have been shown to be inhibitory in nature, we concluded that GABAergic inputs to globus pallidus both at soma and dendrite level are inhibitory. Furthermore, we showed that GABA can promote globus pallidus synchronization by affecting the timing of globus pallidus spiking, and that the globus pallidus GABAergic synapse undergoes rapid frequency-dependent depression. This prominent synaptic depression can account for the ability of globus pallidus neurons to fire in the presence of a majority of inhibitory inputs and might indicate that globus pallidus neurons are tuned to detect frequency changes. Furthermore, globus pallidus synaptic depression rules out the possibility of activation of GABAeregic afferents as the main mechanisms of high-frequency deep brain stimulation, used for treatment of severe parkinsonian patients.

Presynaptic and postsynaptic GABAA receptors in rat suprachiasmatic nucleus.

The mammalian suprachiasmatic nucleus (SCN), the brain's circadian clock, is composed mainly of GABAergic neurons, that are interconnected via synapses with GABA(A) receptors. Here we report on the subcellular localization of these receptors in the SCN, as revealed by an extensively characterized antibody to the alpha 3 subunit of GABA(A) receptors in conjunction with pre- and postembedding electron microscopic immunocytochemistry. GABA(A) receptor immunoreactivity was observed in neuronal perikarya, dendritic processes and axonal terminals. In perikarya and proximal dendrites, GABA(A) receptor immunoreactivity was expressed mainly in endoplasmic reticulum and Golgi complexes, while in the distal part of dendrites, immunoreaction product was associated with postsynaptic plasma membrane. Many GABAergic axonal terminals, as revealed by postembedding immunogold labeling, displayed GABA(A) receptor immunoreactivity, associated mainly with the extrasynaptic portion of their plasma membrane. The function of these receptors was studied in hypothalamic slices using whole-cell patch-clamp recording of the responses to minimal stimulation of an area dorsal to the SCN. Analysis of the evoked inhibitory postsynaptic currents showed that either bath or local application of 100 microM of GABA decreased GABAergic transmission, manifested as a two-fold increase in failure rate. This presynaptic effect, which was detected in the presence of the glutamate receptor blocker 6-cyano-7-nitroquinoxaline-2,3-dione and the selective GABA(B) receptor blocker CGP55845A, appears to be mediated via activation of GABA(A) receptors. Our results thus show that GABA(A) receptors are widely distributed in the SCN and may subserve both pre- and postsynaptic roles in controlling the mammalian circadian clock.

GABA-induced current and circadian regulation of chloride in neurones of the rat suprachiasmatic nucleus.

1. We have shown previously that GABA, the main neurotransmitter in the suprachiasmatic nucleus (SCN), has dual effects on SCN neurones, excitatory during the day and inhibitory at night. This duality has been attributed to changes in [Cl(-)](i) during the circadian cycle. To unravel the processes underlying these changes we investigated the biophysical properties of the GABAergic receptors and the regulation of [Cl(-)](i) in SCN neurones. 2. We used voltage-clamp methodology in conjunction with local application of GABA to characterise the current induced by GABA in SCN neurones within acute brain slices. This current, mediated via GABA(A) receptors, shows moderate voltage dependence, does not desensitise and can significantly alter [Cl(-)](i). 3. Loading or depletion of intracellular Cl(-) was induced by a train of GABA pulses. The recovery of intracellular Cl(-) was deduced from the change in [Cl(-)](i) calculated from the response to a test GABA pulse presented at different intervals after the conditioning train of GABA application. The time course of recovery was described by an exponential curve. Recovery following Cl(-) depletion was slower than recovery from Cl(-) loading and was further delayed during the subjective night. 4. We concluded that: (a) SCN neurones express a large number of somatic GABA(A) receptors, which give rise to a modifiable, tonic Cl(-) conductance that modulates cell excitability; (b) two Cl(-) transport mechanisms operate in SCN neurones, one that replenishes the cell with Cl(-) following Cl(-) depletion and another that removes Cl(-) after Cl(-) loading; (c) the efficiency of the replenishing mechanism is reduced during the subjective night; and (d) this reduction explains a lower [Cl(-)](i) during the night phase of the circadian cycle.

Structural encoding of human and schematic faces: holistic and part-based processes.

The range of specificity and the response properties of the extrastriate face area were investigated by comparing the N170 event-related potential (ERP) component elicited by photographs of natural faces, realistically painted portraits, sketches of faces, schematic faces, and by nonface meaningful and meaningless visual stimuli. Results showed that the N170 distinguished between faces and nonface stimuli when the concept of a face was clearly rendered by the visual stimulus, but it did not distinguish among different face types: Even a schematic face made from simple line fragments triggered the N170. However, in a second experiment, inversion seemed to have a different effect on natural faces in which face components were available and on the pure gestalt-based schematic faces: The N170 amplitude was enhanced when natural faces were presented upside down but reduced when schematic faces were inverted. Inversion delayed the N170 peak latency for both natural and schematic faces. Together, these results suggest that early face processing in the human brain is subserved by a multiple-component neural system in which both whole-face configurations and face parts are processed. The relative involvement of the two perceptual processes is probably determined by whether the physiognomic value of the stimuli depends upon holistic configuration, or whether the individual components can be associated with faces even when presented outside the face context.

Two eyes make a pair: facial organization and perceptual learning reduce visual extinction.

We examined a patient with left spatial neglect and visual extinction due to right parietal damage in tasks where identical stimuli were presented before and after they were primed so as to be perceived as the eyes of schematic faces. In a first block, we presented alphanumeric stimuli (+, o, T, 6) on the right, left, or both sides of fixation on a blank background, and established that the patient could perceive unilateral stimuli on either side but extinguished most of the left-sided ones in the bilateral trials. In a second block, some of these stimuli (+, o) were presented again, but now in the position of eyes within the context of an oval frame which created the impression of a schematic face. Other stimuli (T, 6) were presented as previously on a blank background for an equal number of trials. In the third critical block, all stimuli were presented once again on a blank background, as in the first block. Now the patient extinguished very few of those left-sided stimuli primed to be seen as a pair of eyes in face configuration (+, o), but still extinguished most of the other stimuli (T, 6). A second control experiment showed no effect of repeatedly exposing stimuli in a common region of space defined by meaningless shape boundaries. These results suggest that facial organization can group eye features before the level where attentional selection or extinction occurs, and that such grouping may be influenced by rapid perceptual learning.

Neural fate of seen and unseen faces in visuospatial neglect: a combined event-related functional MRI and event-related potential study.

To compare neural activity produced by visual events that escape or reach conscious awareness, we used event-related MRI and evoked potentials in a patient who had neglect and extinction after focal right parietal damage, but intact visual fields. This neurological disorder entails a loss of awareness for stimuli in the field contralateral to a brain lesion when stimuli are simultaneously presented on the ipsilateral side, even though early visual areas may be intact, and single contralateral stimuli may still be perceived. Functional MRI and event-related potential study were performed during a task where faces or shapes appeared in the right, left, or both fields. Unilateral stimuli produced normal responses in V1 and extrastriate areas. In bilateral events, left faces that were not perceived still activated right V1 and inferior temporal cortex and evoked nonsignificantly reduced N1 potentials, with preserved face-specific negative potentials at 170 ms. When left faces were perceived, the same stimuli produced greater activity in a distributed network of areas including right V1 and cuneus, bilateral fusiform gyri, and left parietal cortex. Also, effective connectivity between visual, parietal, and frontal areas increased during perception of faces. These results suggest that activity can occur in V1 and ventral temporal cortex without awareness, whereas coupling with dorsal parietal and frontal areas may be critical for such activity to afford conscious perception.

Research notes: Fresh and frozen pools of chicken red blood cells as substrates for direct polymerase chain reaction.

A method is presented for reliable use of pooled chicken blood samples for estimation of microsatellite frequencies by direct polymerase chain reaction (PCR) amplification of DNA. This method overcomes the variability of hematocrit values in individual chickens and eliminates the step of DNA preparation. The estimated frequencies of polymorphic alleles in fresh and frozen pooled blood samples were similar to those obtained by calculating these frequencies from the individual genotyping. When frozen pooled blood samples are used, pools should be prepared prior to their freezing.