Robert L. Lloyd

Origin of the neocortically monitored theta rhythm in the curarized rat.

An array of epidural electrodes was acutely implanted in locally anesthetized, curarized rats in order to map the surface distribution of rhythmic slow activity (RSA) which appears within the neocortex. Peak amplitudes (of about 122 muV) were centered over the dorsal hippocampus outline. A laminar profile of RSA recorded within the neocortex indicated no shifts in phase relative to a homotopic, epidural electrode. RSA increased slightly in amplitude (mean increase = 53%) at the deepest level or neocortex, but it did not approximate an amplitude peak or null within the neocortex. Multiple-unit activities within the neocortex were not phase-locked to RSA. On the other hand, all of these manifestations of an RSA generator were observed as electrodes passed into the dorsal hippocampus. A unilateral cortical spreading depression (CSD) treatment, which markedly attenuated barbiturate spindles in all subjects (N = 10), usually (N = 7 of 10) had no effect on the neocortically monitored RSA. Dissociations between depressed and non-depressed hemispheres, and between neocortical and hippocampal RSA, were obtained in some subjects during CSD. However, concurrent dissociations were also apparent between recording sites within the hippocampus. It is concluded that the neocortical RSA of rats is passively spread from the underlying hippocampus, and dissociations in neocortical and hippocampal RSA in the rat are secondary to changes in the organization of multiple generators of hippocampal RSA.

Sensory, motor and cognitive alterations in aged cats

These experiments were designed to assess some of the sensory, motor and cognitive alterations that occur in aged cats. Three groups of cats (1-3, 5-9 and 11-16 years of age) were tested in four behavioral tasks to assess age-dependent changes in locomotor activity, fine motor coordination, reactivity to auditory stimuli and spatial reversal learning. In tests of locomotor activity, 11-16 year old cats displayed altered patterns of habituation compared to 1-3 and 5-9 year cats. There were no decrements in fine motor coordination in the 11-16 year cats as measured by their ability to traverse planks of varying width or by their scores on a neurological examination. The 11-16 and 5-9 year cats both displayed increased reactivity to auditory stimuli. On tests of spatial reversal learning, 11-16 year cats displayed superior performance compared to 5-9 or 1-3 year animals, making fewer errors and requiring fewer trials to reach criterion. These findings indicate that a series of age-related behavioral changes occurs in the cat. Some of these may be related to morphological and neurophysiological alterations in neurons in the caudate nucleus.

Antidepressant effects of thyrotropin-releasing hormone analogues using a rodent model of depression.

The antidepressant potential of two naturally occurring analogues of thyrotropin-releasing hormone (TRH), pGLU-GLU-PRO-NH2 (EEP) and pGLU-PHE-PRO-NH2 (EFP), were examined using a rodent model of antidepressant efficacy. The Porsolt Swim Test was used to assay the antidepressant properties of these two peptides. Both analogues of TRH produced significant antidepressant effects, with EEP producing the stronger response. No effect of EEP upon triiodothyronine (T3) was observed at the dosage used. EFP, which has previously been demonstrated to crossreact with the TRH receptor, significantly increased serum T3. Since an effect upon T3 was only observed in the weaker of the two compounds, these data suggest that the behavioral effect of EEP was not secondary to stimulation of thyroid hormone. Additionally, the differential behavioral response to the two compounds suggests a degree of sequence specificity in the ability of TRH-like tripeptides to produce an antidepressant effect.

TRH Gene Products Are Implicated in the Antidepressant Mechanisms of Seizures

1. After a series of electroconvulsive seizures, levels of TRH-Gly (the immediate precursor of TRH) in four limbic regions correlate significantly and highly with increased swimming in the forced-swim test model of antidepressant efficacy. Only in hippocampus did TRH itself correlate with swimming. 2. After ECS, limbic forebrain regions differ in the relationship of TRH to its precursor peptides. This probably results from differences in the coordination of induction of TRH-processing enzymes, as well as differences in the level of prepro-TRH following seizures. 3. Sprague-Dawley rats that are partially kindled with corneal stimulation swim less in the forced-swim test, opposite to the effect seen with antidepressant agents. 4. Pyriform cortex is unique among the four limbic regions examined in showing decreased amounts of the TRH precursor following swim/stress. 5. Combining ECS with the forced-swim test of antidepressant effects creates a useful model for studying the involvement of TRH and its precursor peptides in both the antidepressant and anticonvulsant effects of controlled therapeutic seizures in the treatment of major depressive disorders. Regional differences between the effects of pinnate and corneal ECS on peptides and behavior support the idea that corneal ECS is a better model than pinnate ECS for human bitemporal ECT. 6. Together with recent results in other laboratories, our results suggest that a series of generalized seizures results in prolonged and increased release and action of TRH in limbic forebrain.

Neurochemical correlates of the Klüver-Bucy syndrome by in vivo microdialysis in monkey

The Klüver-Bucy syndrome is a well known consequence of lesions of the temporal lobe, but the neural mechanisms remain obscure. To elucidate the neurochemical changes in this syndrome, we utilized in vivo microdialysis of amygdala and hypothalamus in two Cebus monkeys (C. apella) before and after bilateral lesions of the temporal pole (TP). Both subjects were housed and observed in a social group when not being dialyzed. Behavioral changes consequent to the TP lesion included early postoperative anorexia, adipsia, hunched posture, tameness, and lethargy. Subsequently loss of fear, hyperorality, loss of social rank, and social withdrawal were observed. Neurochemical changes in amygdala included fall in DA metabolites, increase in NE, and fall in 5-HIAA. The amino acids glutamate and aspartate were both lower postoperatively but more so in the subject with the greatest behavioral changes. Similar changes were noted in hypothalamus except for DA metabolites which remained unchanged. The Klüver-Bucy syndrome consequent to ablation of the temporal pole appears related to a partial deafferentation of excitatory projections to amygdala, along with a lowering of DA and 5-HIAA and an increase in NE.

Neurophysiological alterations in caudate neurons in aged cats.

These neurophysiological studies provide information on the alterations in functional capacity of neurons in the aging caudate nucleus (Cd) of the cat. The major finding is that there is a marked loss of excitation in the Cd during the aging process. This loss is most apparent in animals 11-14 years of age but is demonstrable in animals 6-7 years of age. Extracellular recording techniques were used to test the ability of Cd neurons to respond to activation of two of their major inputs, the precruciate cortex (CX) and the substantia nigra (SN). Types of responses that were evoked in both 1-3- and 11-14-year groups were similar and consisted of excitation, excitation followed by inhibition of action potentials or inhibition alone without preceding excitation. The frequency of occurrence of these responses was altered in the aged animals when either input was stimulated. In 1-3-year-old cats CX stimulation evoked initially excitatory responses in 75% of the cells tested while in 11-14-year-old cats excitatory responses occurred in 62% of the cells. When the SN was stimulated the decrease in initial excitation was greater (69% in 1-3- vs 35% in 11-14-year groups). In all aged animals but not in 1-3-year-old cats stimulation thresholds were higher (39-79%) for evoking excitatory responses than for evoking inhibitory responses. In order to assess synaptic security, the ability of Cd neurons to respond to iterative stimulation was determined. Distributions of the minimum interval necessary to evoke two excitatory responses were constructed. There was a marked increase in the proportion of longer intervals in the aged animals indicating that the synaptic response was less secure. There was a tendency for more of the responses in aged animals to have shorter latencies. This result was probably due to loss of less secure longer latency responses that are mediated via multisynaptic pathways. These findings indicate that there are functional changes in a population of Cd neurons in aged cats that impair their ability to process information.

Electroconvulsive seizures increase levels of pGlu-Glu-Pro-NH2 (EEP) in rat brain

We have previously reported that electroconvulsive seizures (ECS) increases the level of prepro-TRH-derived peptides in hippocampus, amygdala and pyriform cortex but not the striatum of male rats and that this increase is significantly correlated with reduced immobility (increased swimming) in the Porsolt forced swim test. An abstract by Mabrouk and Bennett published in 1993 described increased locomotor activity in rats following IP injection of TRH (pGlu-His-Pro-NH2) and EEP (pGlu-Glu-Pro-NH2). We have examined the effect of three daily transcorneal ECS on the levels of EEP in various brain regions and their correlation with results from the Porsolt forced swim test. The EEP level (ng/g wet weight) was measured by RIA in 6 brain regions: amygdala (AY), hippocampus (HC), pyriform cortex (PYR), anterior cortex (AC), striatum (STR) and motor cortex (MC). ECS significantly increased EEP levels in AY, HC and PYR. The increased swim behavior following ECS, as measured in the Porsolt test, correlated significantly with the EEP levels in HC and MC within individual subjects. Intraperitoneal (IP) injection of EEP (1.0 mg/kg) resulted in a rapid and sustained rise in EEP levels throughout the brain and a clearance half-time from blood of 2.0 h. Intracardiac injection of 0.5 mg EEP resulted in a peak EEP level in CSF at 2 h followed by a t1/2 of 0.35 h. A 3 compartment model for EEP transport from blood into CSF and then brain was developed. This model revealed a 1.75 h delay in the transit time of EEP from blood to CSF followed by rapid clearance from the CSF but long retention time within various brain tissues. We conclude that (1) ECS significantly increases EEP levels in limbic regions, but not in striatum, of the rat brain, (2) EEP, like TRH, is a potential mediator of the antidepressant effect of ECS and (3) EEP, after IP or IV administration, is readily taken up by, and has a long residence time in, brain tissue.

Electroconvulsive seizures increase levels of PS4, the TRH-enhancing peptide [prepro-TRH(160-169)], in rat brain.

We report the development of a radioimmunoassay for prepro-TRH(160-169) (PS4), a thyrotropin-releasing hormone (TRH) enhancing peptide, and its use in characterizing the effect of electroconvulsive seizures on the levels of this peptide in various brain regions of male Wistar rats. We found that electroconvulsive seizures significantly elevated the PS4 levels in hippocampus, amygdala, pyriform (olfactory) cortex, and anterior cortex but not in striatum, motor cortex, locus ceruleus, or ventral lateral medulla. The levels of PS4 were highly correlated with the corresponding TRH (p-Glu-His-Pro-NH2) and TRH-Gly (p-Glu-His-Pro-Gly) levels in hippocampus, amygdala, and pyriform cortex, consistent with the prepro-TRH source of all of these peptides. The PS4 levels in hippocampus and amygdala were significantly correlated with the immobility time in the Porsolt forced swim test, an established animal model for antidepressant effects. The PS4 levels in peripheral blood, hypothalamus, anterior cortex, amygdala, and eyes increased severalfold at 20 min following intracisternal injection of 228 microg of this peptide, suggesting that it readily crosses the blood-brain barrier. The pituitary levels of PS4 and TRH-Gly, on the other hand, were decreased within 20 min by intracisternal PS4, suggesting PS4 stimulated the release of prepro-TRH peptides from the pituitary. Fresh rat and human serum rapidly degraded PS4, indicating that it may act primarily as a paracrine modulator of TRH effects in pituitary, brain, and reproductive system.

Slow wave changes in amygdala to visual, auditory, and social stimuli following lesions of the inferior temporal cortex in squirrel monkey (Saimiri sciureus)

Radiotelemetry of slow wave activity of the amygdala was recorded under a variety of conditions. Power, and the percentage of power in the delta band, increased in response to stimulation. Recordings of monkey vocalizations and slides of ethologically relevant, natural objects produced a greater increase in power than did control stimuli. The responses to auditory stimuli increased when these stimuli were presented in an unrestrained, group setting, yet the responses to the vocalizations remained greater than those following control stimuli. Both the natural auditory and visual stimuli produced a reliable hierarchy with regard to the magnitude of response. Following lesions of inferior temporal cortex, these two hierarchies are disrupted, especially in the auditory domain. Further, these same stimuli, when presented after the lesion, produced a decrease, rather than an increase, in power. Nevertheless, the power recorded from the natural stimuli was still greater than that recorded from control stimuli in that the former produced less of a decrease in power, following the lesion, than did the latter. These data, in conjunction with a parallel report on evoked potentials in the amygdala, before and after cortical lesions, lead us to conclude that sensory information, particularly auditory, available to the amygdala, following the lesion, is substantially the same, and that it is the interpretation of this information, by the amygdala, which is altered by the cortical lesion.

Aging reduces somatosensory responsiveness of caudate neurons in the awake cat

Neuronal activity was recorded in 7 awake cats (3 animals 11-15 years and 4 animals 1-3 years of age) to determine if the ability of caudate neurons to process facial somatosensory information was impaired in aged animals. Proportionately fewer neurons responded to somatosensory stimuli and facial receptive fields were larger in the aged cats. In addition, proportionately fewer caudate neurons responded to cortical activation, there were fewer excitatory responses and spontaneous firing decreasing in aged cats. These results provide additional evidence that excitability of caudate neurons is reduced in aged cats.