F.J. Miñano

Prefrontocortical dopamine depletion induces antidepressant-like effects in rats and alters the profile of desipramine during Porsolt's test.

The objective of this study was to investigate whether bilateral dopamine depletion within the medial prefrontal cortex affects depression state, as well as the antidepressant efficacy of desipramine, in the forced swimming test. The rats behaviour was evaluated by quantifying duration of immobility, climbing, swimming and diving. Immobility latency was also quantified and proved to be a suitable novel parameter. Monoamine levels within the medial prefrontal cortex were measured by high-performance liquid chromatography during Porsolts test, as well as one week after it. While Porsolts test was followed by a typical depression-like profile in sham rats, depletion of prefrontocortical dopamine (86% vs sham controls) reduced immobility and enhanced swimming, which is consistent with a diminished depression tonus. The observed enhancement of swimming was correlated with a high prefrontocortical serotonergic neurotransmission. On the other hand, desipramine induced antidepression-like effects in sham rats by increasing prefrontocortical noradrenaline and serotonin neurotransmisson, but also by blocking the normal increase in dopamine activity during the swimming test. Interestingly, desipramine behaved in a quite different manner in lesioned rats. Thus, immobility duration was not further reduced and only climbing, but not swimming, was enhanced. These effects were correlated with a preferential enhancement of noradrenaline neurotransmission. In conclusion, the results indicate that: (i) dopamine neurotransmission within the medial prefrontal cortex is a factor involved in depression, since dopamine reduction led to a low depression tonus; (ii) desipramine induces antidepression not only by enhancing prefrontocortical noradrenaline and serotonin neurotransmission, but also by blocking the normal increase in dopamine neurotransmission during a depressant situation; (iii) a selective enhancement of prefrontocortical serotonin neurotransmission mediates swimming; and (iv) a selectively augmented prefrontocortical noradrenaline activity mediates climbing during Porsolts test.

Macrophage inflammatory protein-1: Unique action on the hypothalamus to evoke fever

Macrophage inflammatory protein (MIP-1) administered systemically causes a fever not blocked by a prostaglandin (PGE) synthesis inhibitor. The purpose of this study was to examine the central mechanism of pyrexic action of this cytokine in the unrestrained rat. After guide cannulae for microinjection were implanted stereotaxically just above the anterior hypothalamic preoptic area (AH/POA), the body temperature of each rat was monitored by a colonic thermistor probe. Saline control vehicle or MIP-1 was microinjected into the AH/POA in one of eight concentrations ranging from 0.0028-9.0 ng per 0.5 mu 1 volume. MIP-1 induced a biphasic or monophasic fever of short latency characterized by an inverse dose-response curve. The potency of MIP-1 was in the femtomolar (10(-15)) range with the lowest dose of 0.028 ng producing a fever of over 2.0 degrees C with a latency of 15 min or less. To determine whether a PGE mediates MIP-1 fever, indomethacin was administered either intraperitoneally in a dose of 5.0 mg/kg or directly into the MIP-1 injection site in a dose of 0.5 microgram/0.5 mu 1, both injected 15 min before MIP-1. Pretreatment of the injection site in the AH/POA with indomethacin failed to prevent the febrile response evoked by MIP-1 injected at the same locus. Further, the dose of systemic indomethacin, which blocks PGE-induced fever in the rat, attenuated only partially the MIP-1 fever. The results demonstrate that MIP-1 is the most potent endopyrogen discovered thus far, and that its action is directly in the region of the hypothalamus which contains both thermosensitive and pyrogen-sensitive neurons. The local action of MIP-1 on cells of the AH/POA in evoking fever is unaffected by the PGE inhibitor which indicates, therefore, that a cellular mechanism operates in the hypothalamus to evoke fever independently of the central synthesis of a PGE.

GABAA receptors in the amygdala: role in feeding in fasted and satiated rats

The purpose of this study was to clarify further the site of action in the amygdala as well as functional characteristics of feeding in response to two GABA receptor agonists. Guide cannulae for microinjection were implanted stereotaxically in the rat just above the central nucleus of the amygdala (CNA). Microinjections of 0.05, 0.25, 0.5 or 1.0 nmol muscimol, a GABAA-selective receptor agonist, produced a dose- and time-dependent decrease of food intake in both the satiated and fasted rat. The bilateral injection of muscimol into the amygdala was more effective than a unilateral injection during the first 2 h, although the overall effects were similar. Microinjection of 0.1 nmol bicuculline methiodide, a GABAA receptor antagonist, into the CNA significantly blocked this inhibitory effect of 0.05 and 0.5 nmol muscimol again in both the satiated and fasted rat. Doses of 0.05, 0.5, 5.0 and 10.0 nmol of the selective GABAB agonist, baclofen, injected into homologous sites in the CNA did not alter food intake. These findings support the viewpoint that the amygdala and its central nucleus comprise a pivotal region involved in the mechanisms underlying the control of feeding behavior. Further, it is envisaged that hypophagic or anorexic responses are induced through the activation of GABAA receptors by the presynaptic release of GABA from neurons which form a component of the anatomical system for hunger and satiety.

Hypothalamic indomethacin fails to block fever induced in rats by central macrophage inflammatory protein-1 (MIP-1)

This investigation examined the extent to which the activity of a prostaglandin (PG) in the anterior hypothalamic, preoptic area (AH/POA) of the rat plays a role in the intense fever induced by macrophage inflammatory protein-1 (MIP-1) applied directly to this anatomical region. For the microinjection of both a PG synthesis inhibitor, indomethacin, and MIP-1 into sites within the AH/POA, guide cannulae were implanted chronically just above this pyrogen-reactive region. Postoperatively, the body temperature (Tb) of each rat was monitored in the unrestrained condition by means of a colonic thermistor probe. MIP-1 microinjected into the AH/POA in a 0.5-microliter volume evoked a biphasic fever when given in a dose of 5.6 picograms (pg) and a monophasic fever in a dose of 28 pg. The latency of the febrile response was ordinarily 15 min with an asymptote of 1.5 degrees C reached ordinarily within 2.0-2.5 h. When the cytokine-reactive site in the AH/POA was pretreated with indomethacin microinjected in an efficacious dose of 0.5 microgram, the MIP-1 fever evoked by 5.6 pg was not inhibited. Further, pretreatment of AH/POA sites with indomethacin prior to the higher 28-pg dose of MIP-1 delayed the febrile response but did not block it. As a systemic control, indomethacin also was administered intraperitoneally in a dose of 5.0 mg/kg, again 15 min prior to the microinjection of MIP-1 into the AH/POA. In this case, indomethacin only partially attenuated but did not block the fever evoked by either dose of MIP-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Macrophage Inflammatory Protein‐1β (MIP‐1β) Produced Endogenously in Brain During E. coli Fever in Rats

Macrophage inflammatory protein‐1 (MIP‐1) evokes an intense fever, independent of a prostaglandin mechanism, and is now thought to play an important role in the defence response to bacterial pyrogens. The purpose of this study was 2‐fold: (i) to determine whether the potent doublet of this cytokine, MIP‐1β, is actually produced in the brain in response to a pyrogenic dose of a lipopolysaccharide of Escherichia coli and (ii) to determine the anatomical site of synthesis of this cytokine in the brain. Following the intense fever produced by intraperitoneal administration of lipopolysaccharide in the unrestrained rat, MIP‐1β immunoreactivity was identified post mortem in two regions of the brain implicated in fever: the organum vasculosum laminae terminalis (OVLT) and the anterior hypothalamic, preoptic area (AH/POA). Microinjection of goat anti‐mouse MIP‐1β antibody (anti‐MIP‐1β) directly into the AH/POA markedly suppressed fever in rats in response to lipopolysaccharide. Further, anti‐MIP‐1β administered 180 min after the injection of lipopolysaccharide acted as an antipyretic and reversed the fever induced by the endotoxin. Anti‐MIP‐1β or control immunoglobulin G antibody microinjected into the hypothalamus immediately before the intraperitoneal injection of the control saline did not alter the temperature of the rats. Taken together, the present results demonstrate that MIP‐1β is produced in the brain in response to a bacterial endotoxin. These observations, in the light of earlier data on fever induced by MIP‐1β, further support the hypothesis that endogenously synthesized MIP‐1β acts as an intermediary factor in the evocation of fever by acting on the thermosensitive cells of the brain.

Involvement of bicuculline-insensitive receptors in the hypothermic effect of GABA and its agonists

GABA, delta-aminovaleric acid (DAVA) and sodium valproate (VPA) decrease core temperature in conscious rats. Bicuculline increases GABA-induced hypothermia, does not modify DAVA (250 mg/kg) and VPA (100 and 400 mg/kg) hypothermia and antagonizes initial hypothermia by DAVA (1000 mg/kg) and VPA (200 mg/kg) and late hypothermia by DAVA (500 mg/kg) and VPA (200 mg/kg). Picrotoxin increases late hypothermia by GABA (250 mg/kg) and VPA (400 mg/kg), but decreases initial hypothermia by VPA (200 mg/kg). Pentylenetetrazol increases variably GABA-induced hypothermia, inhibits early early hypothermia by DAVA and increases hypothermia induced by VPA (400 mg/kg). We conclude that GABA and GABA-agonists (DAVA and VPA) may induce hypothermia partly mediated by activation of bicuculline-insensitive GABA-receptors.

Circulating Inflammatory Mediators during Start of Fever in Differential Diagnosis of Gram-Negative and Gram-Positive Infections in Leukopenic Rats

ABSTRACT Gram-negative and gram-positive infections have been considered the most important causes of morbidity and mortality in patients with leukopenia following chemotherapy. However, discrimination between bacterial infections and harmless fever episodes is difficult. Because classical inflammatory signs of infection are often absent and fever is frequently the only sign of infection, the aim of this study was to assess the significance of serum interleukin-6 (IL-6), IL-10, macrophage inflammatory protein-2 (MIP-2), procalcitonin (PCT), and C-reactive protein (CRP) patterns in identifying bacterial infections during start of fever in normal and cyclophosphamide-treated (leukopenic) rats following an injection of lipopolysaccharide (LPS) or muramyl dipeptide (MDP) as a model for gram-negative and gram-positive bacterial infections. We found that, compared to normal rats, immunosuppressed animals exhibited significantly higher fevers and lesser production of all mediators, except IL-6, after toxin challenge. Moreover, compared to rats that received MDP, both groups of animals that received an equivalent dose of LPS showed significantly higher fevers and greater increase in serum cytokine levels. Furthermore, in contrast to those in immunocompetent rats, serum levels of IL-6 and MIP-2 were not significantly changed in leukopenic animals after MDP injection. Other serum markers such as PCT and CRP failed to discriminate between bacterial stimuli in both groups of animals. These results suggest that the use of the analyzed serum markers at an early stage of fever could give useful information for the clinician for excluding gram-negative from gram-positive infections.

Hypothalamic interaction between macrophage inflammatory protein-1 alpha (MIP-1 alpha) and MIP-1 beta in rats: a new level for fever control?

1. The microinjection of macrophage inflammatory protein‐1 (MIP‐1 alpha; 5.0 and 25 pg) into the anterior hypothalamic, preoptic area (AHPOA) induced a slow onset; monophasic fever in rats that persisted for a long period. Microinjection of 25 pg MIP‐1 beta into the AHPOA induced a fever of rapid onset, whereas 5.0 pg MIP‐1 beta did not alter body temperature (Tb) significantly. When either MIP‐1 alpha or MIP‐1 beta was heated to 70 degrees C for 30 min prior to their injection, no pyrexic response was produced. 2. The concurrent microinjection of 25 pg MIP‐1 alpha and 25 pg MIP‐1 beta into the AHPOA attenuated the effects on Tb of either cytokine alone. However, pretreatment with either 5.0 pg MIP‐1 beta or 5.0 pg MIP‐1 alpha suppressed the febrile response induced by 25 pg MIP‐1 alpha or 25 pg MIP‐1 beta, given at the same site, respectively. 3. The present experiments show that MIP‐1 alpha and MIP‐1 beta are active individually and possess distinct differences in their evocation of a febrile response. Further, our results suggest a functional antagonism between MIP‐1 alpha and MIP‐1 beta that could represent a new level in the development of fever.

Anorexia and adipsia: Dissociation from fever after MIP-1 injection in ventromedial hypothalamus and preoptic area of rats

Certain cytokines such as tumor necrosis factor (TNF) and interleukin-1 (IL-1) act centrally to affect eating behavior and thermoregulation and may be involved in the physiological mechanisms leading to anorexia, adipsia and loss in body weight. The newly discovered macrophage inflammatory protein-1 (MIP-1) infused into the anterior hypothalamic, preoptic area (AH/POA) evokes an intense hyperthermia. The present experiments were designed to determine whether MIP-1 affects the feeding mechanism in the ventromedial hypothalamus (VMH) independently of the thermoregulatory mechanism in the AH/POA. For the microinjection of MIP-1, guide cannulae were implanted stereotaxically in the rat just above the VMH or AH/POA. Following postoperative recovery, each unrestrained rat was adapted to procedures whereby body temperature and intakes of food and water available ad lib were monitored at predetermined intervals. When an efficacious dose of 5.6 picograms (pg) MIP-1 was microinjected in a volume of 0.5 microliters into the VMH, the intake of food in the rat was reduced significantly in the short term and throughout the following 22 h. Within intervals of 30 min and 4.0 h following MIP-1, the amount of food consumed was 4.0 and 10 g, respectively, below that eaten by control rats given the saline solvent vehicle injected at the same site in the VMH. Over the entire test period, the intake of water was similarly significantly below that of the control rats. Whereas MIP-1 injected into the AH/POA evoked fever accompanied by a transient decline in feeding, the body temperature of the rats was unaffected by the cytokine injected in the VMH.(ABSTRACT TRUNCATED AT 250 WORDS)

Opioid and prostaglandin mechanisms involved in the effects of GABAergic drugs on body temperature.

1. Intraperitoneal (i.p.) injection to restrained rats of GABA (250-1000 mg/kg) or the GABAA-receptor agonist muscimol (0.05-1 mg/kg) induced a dose-dependent decrease in body temperature (BT). 2. Intraperitoneal injection of low doses of the GABAB-receptor agonist (+/-)-baclofen (1-10 mg/kg) did not significantly affect BT. However, baclofen, at high doses (30 mg/kg), produced an increase in BT. 3. Pretreatment with either bicuculline (3 mg/kg) or naloxone (1 mg/kg) did not significantly modify the hypothermic response observed with GABA or muscimol, except for the high dose of GABA (1000 mg/kg) which was potentiated by bicuculline pretreatment. 4. Indomethacin pretreatment (5 mg/kg) significantly antagonized the hypothermia induced by GABA and muscimol. 5. Injection of baclofen alone (1 mg/kg) did not significantly affect BT, but in presence of the GABAA antagonist bicuculline, baclofen significantly decreased BT. 6. Baclofen-induced hyperthermia appear to be via prostaglandin and opioid mechanisms since both indomethacin and naloxone abolish this effect. 7. The hypothermia induced by GABA-agonists appears to be due to simultaneous activation of GABAA and GABAB receptors, while the hyperthermic effect of baclofen appears to be due to stimulation of GABAB receptors. 8. The present results suggest that involvement of prostaglandins in the effects of GABA, muscimol and baclofen, while endogenous opiates seem to be implicated only in baclofen induced hyperthermia. 9. It can be concluded that GABA may be involved in the control of BT through GABAA and GABAB receptors.