Francisco Gonzalez-Lima

Prefrontal Mechanisms in Extinction of Conditioned Fear

Interest in the medial prefrontal cortex (mPFC) as a source of behavioral inhibition has increased with the mounting evidence for a functional role of the mPFC in extinction of conditioned fear. In fear extinction, a tone-conditioned stimulus (CS) previously paired with a footshock is presented repeatedly in the absence of footshock, causing fear responses to diminish. Here, we review converging evidence from different laboratories implicating the mPFC in memory circuits for fear extinction: (1) lesions of mPFC impair recall of extinction under various conditions, (2) extinction potentiates mPFC physiological responses to the CS, (3) mPFC potentiation is correlated with extinction behavior, and (4) stimulation of mPFC strengthens extinction memory. These findings support Pavlovs original notion that extinction is new learning, rather than erasure of conditioning. In people suffering from posttraumatic stress disorder (PTSD), homologous areas of ventral mPFC show morphological and functional abnormalities, suggesting that extinction circuits are compromised in PTSD. Strategies for augmenting prefrontal function for clinical benefit are discussed.

Antidepressant-like effects of medial prefrontal cortex deep brain stimulation in rats.

BACKGROUND Subcallosal cingulate gyrus (SCG) deep brain stimulation (DBS) is being investigated as a treatment for major depression. We report on the effects of ventromedial prefrontal cortex (vmPFC) DBS in rats, focusing on possible mechanisms involved in an antidepressant-like response in the forced swim test (FST). METHODS The outcome of vmPFC stimulation alone or combined with different types of lesions, including serotonin (5-HT) or norepineprhine (NE) depletion, was characterized in the FST. We also explored the effects of DBS on novelty-suppressed feeding, learned helplessness, and sucrose consumption in animals predisposed to helplessness. RESULTS Stimulation at parameters approximating those used in clinical practice induced a significant antidepressant-like response in the FST. Ventromedial PFC lesions or local muscimol injections did not lead to a similar outcome. However, animals treated with vmPFC ibotenic acid lesions still responded to DBS, suggesting that the modulation of fiber near the electrodes could play a role in the antidepressant-like effects of stimulation. Also important was the integrity of the serotonergic system, as the effects of DBS in the FST were completely abolished in animals bearing 5-HT, but not NE, depleting lesions. In addition, vmPFC stimulation induced a sustained increase in hippocampal 5-HT levels. Preliminary work with other models showed that DBS was also able to influence specific aspects of depressive-like states in rodents, including anxiety and anhedonia, but not helplessness. CONCLUSIONS Our study suggests that vmPFC DBS in rats may be useful to investigate mechanisms involved in the antidepressant effects of SCG DBS.

Structural modeling of functional neural pathways mapped with 2-deoxyglucose: effects of acoustic startle habituation on the auditory system

This paper describes the first application of structural modeling to neuroscience. Structural modeling (also known as path analysis) is a method to assess the relative impact of directional links in a system and how these interrelations may change under different conditions. The objective was to demonstrate how structural modeling can be used to determine the functional interrelationships between brain structures that form the auditory system. Using structural modeling, changes in auditory system 2-DG uptake were examined during long- and short-term habituation of the acoustic startle reflex. Models were based on the anatomical connections between central auditory system structures. Using functional 2-DG data, the correlations between these structures were calculated and numerical weights were computed for each anatomical link. The analysis revealed that the lemniscal path was dominant during short-term habituation, while during long-term habituation this influence was modified through extra-lemniscal pathways. The models are discussed in the context of previous findings to demonstrate how structural modeling can not only complement, but also extract more information from 2-DG mapping experiments.

Neural substrates for tone-conditioned bradycardia demonstrated with 2-deoxyglucose. II. Auditory cortex plasticity

The 2-deoxyglucose (2-DG) method was used to map the metabolic activity of the auditory cortex (AC) during and after conditioning. Using separate groups of animals, the effects of both paired and unpaired presentations of a 4-5 kHz FM tone (CS) and midbrain reticular stimulation (US) were compared for acquisition, extinction and sensitization training. Rats with cardiac deceleration conditioned to the FM tone showed a pattern of AC metabolic activity distinctly different from that seen in control animals. The tonotopic pattern of 2-DG labeling consisted of two contiguous spindle-shaped bands corresponding to the location of neurons with best frequency response in the 4-5 kHz band width. Reticular stimulation alone or combined with the tones produced a widespread increase of 2-DG uptake. At least two types of modulatory effects appeared to interact with the tonotopic pattern. The first involved a selective enhancement of evoked activity in the AC region of convergence of CS-US effects. This effect may be related to learning because it was restricted to the conditioning group. The second effect involved a general increase in background uptake of 2-DG in AC. This effect may be related to reticular sensitization because it was common to all groups subjected to reticular stimulation. The present findings are the first anatomical demonstration of the modulatory effects of auditory learning on AC metabolic activity.

Functional Connectivity among Limbic Brain Areas: Differential Effects of Incubation Temperature and Gonadal Sex in the Leopard Gecko, Eublepharis macularius

The neural basis of individual differences in behavior has been studied primarily by analyzing the properties of specific neural areas. However, because of the organization of the nervous system, it is also plausible that differences in behavior are mediated by differences in the interactivity or functional connectivity among brain nuclei in particular neural circuits. In the leopard gecko, Eublepharis macularius, the temperature of the egg during incubation not only determines gonadal sex, but also shapes the sociosexual behaviors, reproductive physiology, and hormone sensitivity of adult animals. In this study the effects of both incubation temperature and gonadal sex on functional connectivity among limbic brain areas were examined. Functional connectivity was assessed by analyzing covariance patterns in metabolic capacity, as revealed by quantitative cytochrome oxidase histochemistry. It was hypothesized that incubation temperature and gonadal sex affect the propensity to display aggressive or sexual behaviors by altering the functional connectivity within relevant neural circuits. The correlations of metabolic capacity between the anterior hypothalamus and both the septum and preoptic area were significant only in relatively aggressive individuals, suggesting that these circuits may regulate the phenotypic variation in aggressiveness caused by incubation temperature. The correlations between the ventromedial hypothalamus and both the dorsal ventricular ridge and septum were significant only in females, suggesting that these circuits may modulate female-typical sexual behaviors. Correlations among preoptic, hypothalamic and amygdalar areas tended to be distributed across both sexes, suggesting that there may be shared pathways underlying the expression of male-typical and female-typical behaviors.

Quantitative histochemistry of cytochrome oxidase in rat brain

A quantitative analysis of cytochrome oxidase (CO) activity in histochemically stained sections of rat brain was developed using tissue standards and computerized image processing. Standards of brain paste containing known amounts of CO were cryosectioned and stained under the same conditions as brain sections. The gray levels of the stain were converted to units of CO activity using a calibration curve derived from densitometric analysis of the standards. The technique yields reproducible quantitative values, has the superior anatomical resolution of histochemistry, and is compatible with autoradiography.

Cytochrome oxidase in neuronal metabolism and Alzheimer's disease

Neuronal Metabolism and Cytochrome Oxidase Function: Brain Cytochrome Oxidase: Functional Significance and Bigenomic Regulation in the CNS M.T.T. Wong-Riley, et al. Quantitative Histochemistry of Cytochrome Oxidase Activity: Theory, Methods, and Regional brain Vulnerability F. Gonzalez-Lima, A. Cada. Cytochrome Oxidase and Neuroanatomical Patterns: What is the Connection? R.F. Hevner. Functional Mapping of Learning-Related Metabolic Activity with Quantitative Cytochrome Oxidase Histochemistry A. Poremba, et al. Functional Imaging Probes to Study the Neural Bases of Behavior in Genetic Animal Models of ADHD: A Comparative Analysis of Short and Long-Term Markers of Neuronal Activity M. Papa, et al. Alzheimers Disease and Cytochrome Oxidase Dysfunction: Cytochrome Oxidase Inhibition in Alzheimers Disease F. Gonzalez-Lima, et al. Molecular Mechanisms of Impaired Mitochondrial Gene Expression in Alzheimers Disease K. Chandrasekaran, et al. Behavioral, Electrophysiological, and Biochemical Consequences of Chronic Cytochrome Oxidase Inhibition in Rats M.C. Bennett, G.M. Rose. Cytochrome Oxidase: A Predictive Marker of Neurodegeneration N.P. Abdollahian, et al. Appendix. Cytochrome Oxidase Atlas of Rat Brain F. Gonzalez-Lima, A. Cada. Index.

Behavioral effects of metyrapone on Pavlovian extinction

This is the first study of the action of metyrapone on Pavlovian extinction. Pavlovian acquisition memory can be impaired when 50 mg/kg metyrapone, a corticosterone synthesis inhibitor, is injected 90 min before training. It was hypothesized that the same treatment given before extinction may also impair Pavlovian extinction memory, and thereby facilitate recovery of the extinguished behavior. This study examined the behavioral effects of 50 mg/kg metyrapone on the extinction of conditioned freezing following Pavlovian conditioning of tone (CS) and footshock (US). On days 1-2, mice were habituated to the training context. On days 4-5, mice received 4 tone-shock pairings per day. On day 6, metyrapone or saline was injected s.c. 90 min before an extinction session with 60 tone presentations. Probe sessions with 4 tones were conducted in the extinction context on day 7 and in the acquisition context on day 9. Metyrapone treatment did not affect performance during extinction or pre-CS freezing behavior. But metyrapone-treated animals showed greater conditioned freezing when tested with the tone the day after extinction in the extinction context (spontaneous recovery) and 3 days after extinction in the acquisition context (renewal effect). It was concluded that 50 mg/kg metyrapone did not affect extinction performance, but it effectively facilitated the subsequent recovery of the extinguished behavior. This effect may be explained by an impairment of the consolidation of the Pavlovian extinction memory. This interpretation is consistent with previous studies showing that metyrapone may interfere with memory consolidation for a variety of learned responses.

Classical conditioning of tone-signaled bradycardia modifies 2-deoxyglucose uptake patterns in cortex, thalamus, habenula, caudate-putamen and hippocampal formation

The 2-[14C]deoxyglucose (2-DG) autoradiographic method was used to map metabolic activity in all telencephalic and diencephalic structures of the rat brain during and after classical conditioning. A trial was made of a 4-5 KHz frequency modulated tone (CS) paired with midbrain reticular stimulation (US). The unconditioned response was a rapid bradycardia elicited by the US. Alert rats were injected with 2-DG, placed in a sound-proof chamber, and subjected during 90 min to a given treatment: (1) the CS before conditioning, (2) the US alone, (3) the paired CS-US (acquisition), (4) the CS after conditioning (extinction), (5) the US prior to the CS (sensitization), (6) the unpaired CS-US (pseudoconditioning), (7) the CS after pseudoconditioning and (8) no stimulation. The prefrontal cortex showed discrete regions with enhanced 2-DG uptake during conditioning and pseudoconditioning. A columnar organization was well-defined in the posterior parietal cortex of rats subjected to CS-US pairing. The medial thalamus was greatly activated in all groups subjected to reticular stimulation. The dorsomedial nucleus showed its largest activation during conditioning. The lateral habenula and a caudal portion of caudate-putamen showed an overall increase in 2-DG uptake during conditioning. The hippocampal formation showed a specific pattern of metabolic activation during conditioning and after conditioning. A laminar densitometric analysis showed that 2-DG uptake was concentrated in a central band along the sides of the hippocampal fissure which corresponded to the molecular layers. Only this neuropil band of greater metabolic activity showed the learning-related changes. In addition, the hippocampal formation was the only nonauditory structure in the forebrain which clearly responded to the acquired signal value of the tone CS after conditioning. These changes revealed by 2-DG provide a first demonstration of forebrain substrates with localized metabolic alterations related to learning and reticular sensitization.

In Vivo Low‐level Light Therapy Increases Cytochrome Oxidase in Skeletal Muscle

Low‐level light therapy (LLLT) increases survival of cultured cells, improves behavioral recovery from neurodegeneration and speeds wound healing. These beneficial effects are thought to be mediated by upregulation of mitochondrial proteins, especially the respiratory enzyme cytochrome oxidase. However, the effects of in vivo LLLT on cytochrome oxidase in intact skeletal muscle have not been previously investigated. We used a sensitive method for enzyme histochemistry of cytochrome oxidase to examine the rat temporalis muscle 24 h after in vivo LLLT. The findings showed for the first time that in vivo LLLT induced a dose‐ and fiber type‐dependent increase in cytochrome oxidase in muscle fibers. LLLT was particularly effective at enhancing the aerobic capacity of intermediate and red fibers. The findings suggest that LLLT may enhance the oxidative energy metabolic capacity of different types of muscle fibers, and that LLLT may be used to enhance the aerobic potential of skeletal muscle.