Time factors in conscious processes: reply to Gilberto Gomes.

The critical reinterpretations of Libet's research by G. Gomes make speculative, unwarranted, and untested assumptions. These assumptions and arguments are analyzed and their status relative to Libet's findings is criticized.

The human locus coeruleus and anxiogenesis.

Electrical stimulation of locus coeruleus (LC), via permanently implanted electrodes with confirmed localization and effectiveness, did not elicit any subjective or behavioral manifestations of anxiety. This is evidence against the hypothesis that LC is a mediator of anxiogenesis in man.

The neural time factor in conscious and unconscious events.

Our earlier evidence had indicated that a substantial duration of appropriate cerebral activity (up to about 0.5 s) is required for the production of a conscious sensory experience; this means the sensory world is experienced delayed with respect to real time. Subjective timing of the experience can be retroactively referred to the time of the earliest signal arriving at the cortex. Our 'time-on' theory states that the transition from an unconscious to a conscious mental function is determined, at least in part, by an increase in the duration of appropriate neural activities. Our experimental finding that conscious intention to act appears only after a delay of about 350 ms from the onset of specific cerebral activity that precedes a voluntary act provided indirect evidence for the theory. In a direct experimental test a signal (stimulus to somatosensory thalamus) was correctly detected in a forced-choice test even when the stimulus duration was too short to produce any awareness of the signal; to go from correct detection with no awareness to detection with awareness required an additional 400 ms of the repetitive identical neural volleys ascending to sensory cortex. 'Time-on' theory has important implications for a variety of unconscious-conscious interactions.

Introduction to slow synaptic potentials and their neuromodulation by dopamine.

The existence of two muscarinically mediated slow postsynaptic potentials (PSPs) and a noncholinergic (peptidergic) late-slow PSP was established in the 1960s. These have synaptic delays and PSP durations 100-10,000 times those for the nicotinic (fast) excitatory post-synaptic potential (EPSP). Evidence is reviewed for an against the proposal that, in rabbit superior cervical ganglia, the slow (s-) inhibitory postsynaptic potential requires a second transmitter, dopamine, released by muscarinic action on interneurones (the small, intensely fluorescent cells). The s-EPSP in frog ganglia appears only in already depolarized cells by a muscarinic closure of the M (voltage-sensitive K+) channels. But the large s-EPSP in mammalian neurones, not depolarized, is generated largely via other mechanisms, especially one involving cyclic GMP. Dopamine also produces a long-term enhancement (LTE) of the muscarinic slow PSPs in rabbit superior cervical ganglia, whether dopamine is applied exogenously or released intraganglionically by preganglionic nerve impulses at 10 s-1. LTE is producible heterosynaptically, and it persists well over 3 h; a noncholinergic (peptide?) transmitter may contribute to the initial 30 min of LTE. LTE is mediated by a D1 receptor coupled to cyclic AMP; it is blocked by cyclic GMP or low Ca2+ or calmidazolium (a calmodulin inhibitor). The modulatory process of LTE has certain similarities to, but also fundamental differences from, the long-term potentiation known in the hippocampus.

Control of the transition from sensory detection to sensory awareness in man by the duration of a thalamic stimulus. The cerebral 'time-on' factor.

A 'time-on' theory to explain the cerebral distinction between conscious and unconscious mental functions proposes that a substantial minimum duration ('time-on') of appropriate neuronal activations up to about 0.5 s is required to elicit conscious sensory experience, but that durations distinctly below that minimum can mediate sensory detection without awareness. A direct experimental test of this proposal is reported here. Stimuli (72 pulses/s) above and below such minimum train durations (0-750 ms) were delivered to the ventrobasal thalamus via electrodes chronically implanted for the therapeutic control of intractable pain. Detection was measured by the subject's forced choice as to stimulus delivery in one of two intervals, regardless of any presence or absence of sensory awareness. Subjects also indicated their awareness level of any stimulus-induced sensation in each and every trial. The results show (1) that detection (correct greater than 50%) occurred even with stimulus durations too brief to elicit awareness, and (2) that to move from mere detection to even an uncertain and often questionable sensory awareness required a significantly larger additional duration of pulses. Thus simply increasing duration ('time-on') of the same repetitive inputs to cerebral cortex can convert an unconscious cognitive mental function (detection without awareness) to a conscious one (detection with awareness).

Postsynaptic long-term enhancement (LTE) by dopamine may be mediated by Ca2+ and calmodulin.

Long-term enhancement (LTE), of postsynaptic slow depolarizing responses to a muscarinic agonist (MCh), follows a brief exposure of the rabbit superior cervical ganglion to another transmitter, dopamine (DA). Either reduction of external Ca2+ (to 1.0 mM or 0.2 mM) or presence of a specific calmodulin antagonist (calmidazolium at 5 microM) blocked DA induction of this LTE. However, unlike LTP in hippocampus, induction of LTE is not mediated by depolarization-dependent influx of Ca2+.

Stimulation of locus coeruleus in man. Preliminary trials for spasticity and epilepsy.

Stimulating electrodes were chronically implanted unilaterally (in 1975-1977) in the vicinity of the locus coeruleus (LC) in three patients, one with cerebral palsy-spastic quadriplegia, two with epilepsy (one grand mal, one psychomotor). Effective excitation of efferent LC axons was indicated by measuring rises in 3-methoxy-4-hydroxyphenylethyleneglycol in the jugular and systemic venous blood following a 6-min stimulus with discontinuous bursts of pulses. There was a substantial reduction of spasticity during and after stimulation. Improvement was verified by double-blind failures of the stimulator, and the stimulus therapy is still in use after 9 years. There appeared to be a reduction in incidence and severity of both types of epileptic seizures, although this was not rigorously established. The patient with psychomotor epilepsy reported a considerable lengthening of preseizure auras (to 15-30 min), an unusual number of which terminated without a seizure.

Long-term enhancement (LTE) of postsynaptic potentials following neural conditioning, in mammalian sympathetic ganglia.

Orthodromic, preganglionic conditioning stimulation can consistently induce long-term enhancement (LTE) (greater than 3 h) of the muscarinically mediated slow excitatory postsynaptic potential and the slow inhibitory postsynaptic potential. This was shown for superior cervical ganglia of rabbit and rat. Effective conditioning stimuli are in a physiologically observed range (3/s for 7 min, 5/s for 4 min, 10/s for 2 min, 20/s for 1 min). LTE was producible both homosynaptically and heterosynaptically. LTE can thus be associative, with conditioning synaptic input in one line inducing long-term changes in postsynaptic responses to another (heterosynaptic) input. The dopamine antagonist butaclamol depressed LTE, particularly that following the initial postconditioning period of 30 min. Adrenergic antagonists had no effect. This pharmacological evidence, coupled with the heterosynaptic induction of LTE, supports the view that neurally induced LTE may be at least partly mediated by endogenous dopamine. Another non-cholinergic but non-adrenergic transmitter (possibly a peptide) might contribute to the LTE seen in the initial 30 min postconditioning. The present, orthodromically induced LTE is clearly different from the long-term potentiation widely studied in hippocampus, etc., in the modes of induction and synaptic mediation.

Secondary late components of the muscarinic postsynaptic potentials, in rabbit superior cervical ganglion.

The well known muscarinic slow excitatory polysynaptic potential (s-EPSP) of rabbit superior cervical ganglion (SCG) peaking at about 1-2 s and lasting 5-10 s, is immediately followed by an abrupt change in slope to a longer, lower depolarizing phase. A brief dip in the level of depolarization (DP) often separates the two depolarizing phases. The secondary phase of s-EPSP rises to its own peak at about 25 s; total duration 60-120 s. With repetition of orthodromic volleys secondary s-EPSP builds up more gradually than initial s-EPSP, but more rapidly than slow-slow (ss-) EPSP. The later 'secondary' depolarizing phase along with the antecedent 'dip in DP' are, like the 'initial' s-EPSP, eliminated by a muscarinic antagonist, quinuclinidyl benzilate hydrochloride (QNB). This distinguishes secondary s-EPSP from the even slower rising non-cholinergic ss-EPSP. The ss-EPSP, although relatively small in the responses to the usual 3-pulse test stimuli, rises to an extraordinary amplitude (equal to the compound action potential) during a 10 s-120 s train of pulses. Gallamine blocked most of the slow IPSP component in test responses but not initial or secondary s-EPSP. A preganglionic conditioning train (10/s for 2 min) induced a long-term-enhancement (LTE) of secondary s-EPSP lasting greater than 3 h, with maximum postconditioning percentage increases greater than for initial s-EPSP. Also enhanced was the dip in DP, now forming a deeper notch between initial and secondary s-EPSPs; this attains a maximum at about 30 min postconditioning but thereafter progressively loses the enhancement by about 90 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Stimulation of adenylate cyclase in relation to dopamine-induced long-term enhancement (LTE) of muscarinic depolarization in the rabbit superior cervical ganglion.

Dopamine (DA) induction of the long-term enhancement (LTE) of the slow muscarinic depolarizing response to methacholine (MCh), equivalent to the slow EPSP (S-EPSP), was previously found to be mimicked by exogenous cyclic AMP (cAMP) in the rabbit superior cervical ganglion (SCG). DA-induced LTE of the S-EPSP was shown to be depressed by some DA antagonists. We now show that DA (15 microM), its analog, 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (ADTN), and a D2 receptor antagonist, metoclopramide, each can induce both LTE of MCh depolarization and an increase in ganglionic cAMP. Conversely, antagonists of DA-induced LTE also depress DA-induced rises in cAMP; these antagonists include haloperidol (1 microM), both (+) and (-) enantiomers of butaclamol (0.7-7 microM), flupenthixol (1 microM), and (+)-R-8-chloro-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1H-3-benzazepine-7-o l (SCH-23390) (7 microM). The selective D2 antagonists sulpiride (10 microM) and domperidone (10 microM) affect neither DA action. Alpha-2 adrenergic agonists (alpha-methyl-norepinephrine and clonidine) produce no LTE; alpha-antagonist dihydroergotamine (35 microM) does not affect either DA action, although it can completely block the hyperpolarizing response to DA or other catecholamines. Beta-antagonist propranolol (5 microM) partially depresses DA-induced rises in cAMP but has no effect on the DA-induced LTE. (Butaclamol and propranolol in combination can completely block the cAMP rise induced by DA.) Beta-agonist isoproterenol can induce appreciable LTE of MCh depolarization, but this LTE is not depressed by propranolol (10 microM). Isoproterenol can elicit a substantial rise in cAMP.(ABSTRACT TRUNCATED AT 250 WORDS)

Nonclassical synaptic functions of transmitters.

Nonclassical synaptic functions are considered in two groups, mainly by reference to the models provided by sympathetic ganglia. Slow postsynaptic potentials (PSPs) are compared with classical fast PSPs. Features include loose delivery of transmitter to receptor, very long synaptic delays and durations of PSPs, slow removal of transmitter or of its effects, integration of repetitive inputs, electrogenesis without large increases in ionic conductances. Neuromodulatory actions affect synaptic efficacy without direct excitatory or inhibitory responses to the transmitter. These include a) control of presynaptic release, and b) contingent postsynaptic actions. In b, a modulatory transmitter alters the efficacy of action by another transmitter. The alteration may persist long after exposure to the modulatory transmitter; in mammalian sympathetic ganglia, exposure to dopamine or to a conditioning train of preganglionic volleys induces a long-term enhancement of the muscarinic slow excitatory PSP. Or the alteration may be restricted mostly to the presence of a modulatory transmitter, with examples cited. Nonclassical synaptic functions may be providing revolutionary possibilities for dealing with slow and broadly distributed cerebral functions, manifested electrophysiologically and behaviorally, that have been difficult to analyze successfully in terms restricted to the fast and discretely localized classical synaptic functions.