Madeline L. Cooper

Roles of gender, gonadectomy and estrous phase in the analgesic effects of intracerebroventricular morphine in rats

Gender and gonadal function have previously been shown to influence the magnitude of analgesia following systemic morphine and opioid and nonopioid forms of swim analgesia with male rats displaying greater analgesia than female rats and gonadectomy reducing analgesic magnitude in both genders. These effects have been presumed to be centrally mediated. The present study evaluated the roles of gender, gonadectomy and estrous phase upon dose-response and time-response functions of analgesia following intracerebroventricular administration of morphine as measured by the tail-flick and jump tests. Sham-operated male rats displayed significantly greater magnitudes of peak and total analgesia following central morphine than sham-operated female rats on both nociceptive measures. This striking effect was reflected both in terms of magnitude and ED50; while male rats displayed near-maximal analgesia at a 5 micrograms dose of morphine, female rats displayed moderate analgesia at doses as high as 40 micrograms of morphine. Castration produced small, but significant reductions in the magnitude of central morphine analgesia; the ED50 of morphine analgesia, however, was not changed. Although female rats in either proestrous or estrous displayed significantly greater magnitude of analgesia than ovariectomized rats or rats in a combined met-/di-estrous phase at some doses, the ED50 of morphine analgesia was not significantly altered as functions of estrous phase or ovariectomy. The interaction of opiate receptors and gonadal steroid receptors is considered as one possible determinant of gender differences observed in the magnitude and potency of central morphine analgesia.

Interactions among aging, gender, and gonadectomy effects upon morphine antinociception in rats.

In addition to age-related deficits in morphine antinociception in female rats, gender and gonadectomy differences have also been observed, with male rats displaying greater magnitudes of effects than females and castrated males. Since there are little data indicating how aging, gender, and gonadectomy interact in modulating morphine antinociception, the present study evaluated alterations in this response as functions of age (6, 12, 18, and 24 months), gender, and gonadal status (intact, gonadectomized) across a dose range (1-10 mg/kg) and time course (0.5-2 h) on the tail-flick test. The maximal percentage effect (MPE) of morphine (1 mg/kg) was significantly increased in castrated males (18 months), sham females (18 and 24 months), and ovariectomized females (18 months) relative to 6-month-old groups. Increases in the MPE of morphine (1 mg/kg) occurred in sham females (24 months) relative to corresponding sham males and ovariectomized females. The MPE of morphine (2.5 mg/kg) was significantly increased in sham males (18 months) and decreased in sham females (12 months). Decreases in the MPE of morphine (2.5 mg/kg) occurred in castrated males (18 and 24 months) as well as sham (18 months) and ovariectomized (18 and 24 months) females relative to sham males. Whereas the MPE of morphine (5 mg/kg) was unchanged by these variables, the MPE of morphine (10 mg/kg) was significantly decreased in sham females (18 and 24 months) relative to females aged 6 months, as well as males and ovariectomized females aged 24 months.(ABSTRACT TRUNCATED AT 250 WORDS)

General, μ and κ opioid antagonists in the nucleus accumbens alter food intake under deprivation, glucoprivic and palatable conditions

Abstract Ventricular microinjection studies found that whereas μ (β-funaltrexamine, B-FNA), μ1 (naloxonazine) and κ (nor-binaltorphamine, Nor-BNI) opioid receptor antagonists, but not δ antagonists, reduce deprivation-induced intake, κ and μ, but not μ1 or δ antagonists reduce both 2-deoxy- d -glucose (2DG) hyperphagia and sucrose intake. Since opioid agonists stimulate spontaneous food intake in the accumbens, the present study examined whether administration of either naltrexone, B-FNA or Nor-BNI in the accumbens altered intake under deprivation (24 h), glucoprivic (2DG: 500 mg/kg, i.p.) or palatable sucrose (10%) conditions. Naloxonazines effects in the accumbens were also evaluated for deprivation-induced intake. Deprivation-induced intake was significantly decreased over 4 h by naltrexone (5–20 μg, 44%), B-FNA (1–4 μg, 55%) and Nor-BNI (4 μg, 31%), but not naloxonazine (10 μg) in the accumbens. 2DG hyperphagia was significantly decreased by naltrexone (10–20 μg, 79%), B-FNA (1–4 μg, 100%) and Nor-BNI (1–4 μg, 75%) in the accumbens. Sucrose intake was significantly decreased by naltrexone (50 μg, 27%) and B-FNA (1–4 μg, 37%), but not Nor-BNI in the accumbens. These data suggest that μ receptors, and particularly the μ2 binding site in the accumbens are responsile for the opioid modulation of these forms of intake in this nucleus, and that this control may be acting upon the amount of intake per se.

Modulation of gender-specific effects upon swim analgesia in gonadectomized rats

Gender-specific effects have been observed for continuous and intermittent cold-water swim (CCWS and ICWS respectively) analgesia: analgesic magnitudes following CCWS and ICWS are significantly smaller in female rats than in age-matched and weight-matched male rats. The present study evaluated the role of gonadal status in these gender-specific effects by examining CCWS and ICWS analgesia, hypothermia and activity in intact and gonadectomized rats. Following confirmation of the original gender-specific effects on the tail-flick and jump tests, it was found that both castration and ovariectomy significantly reduced CCWS and ICWS analgesia. Indeed, castrated males displayed similar magnitudes of analgesia as intact females. The more marked hypothermia observed in intact females indicated that this variable failed to account for the analgesic gender-specific effects. The reduced hypothermia following gonadectomy also failed to account for the analgesic changes. The increased activity during ICWS, but not CCWS following gonadectomy also did not account for the analgesic changes. These data suggest that gonadal steroids normally appear to facilitate these stress-related analgesic responses.

Excitatory amino acid antagonists in the rostral ventromedial medulla inhibit mesencephalic morphine analgesia in rats

&NA; Supraspinal opioid analgesia is mediated in part by connections between the periaqueductal gray (PAG) and rostral ventromedial medulla (RVM). Morphine analgesia elicited from the PAG is respectively decreased by selective serotonergic and opioid receptor antagonists administered into the RVM, and increased by RVM neurotensin antagonists. Since glutamate and excitatory amino acid (EAA) receptors are also active in the RVM, the present study evaluated whether either competitive (AP7) or non‐competitive (MK‐801) N‐methyl‐D‐aspartate (NMDA) antagonists or a kainate/AMPA (CNQX) antagonist microinjected into the RVM altered morphine (2.5 &mgr;g) analgesia elicited from the PAG as measured by the tail‐flick and jump tests. Mesencephalic morphine analgesia was markedly reduced on both tests after RVM pretreatment with either AP7 (0.01‐1 &mgr;g, 0.08‐7.8 nmol) or MK‐801 (0.03‐3 &mgr;g, 0.04‐4.4 nmol). In contrast, small but significant reductions in mesencephalic morphine analgesia occurred on the jump test following CNQX (0.5 &mgr;g, 2.2 nmol) in the RVM. NMDA antagonists did not markedly alter either basal nociceptive thresholds following RVM administration, or mesencephalic morphine analgesia following administration into medullary placements lateral or dorsal to the RVM. These data implicate EAA and particularly NMDA receptors in the RVM in modulating the transmission of opioid pain‐inhibitory signals from the PAG.

Opioid antagonists in the periaqueductal gray inhibit morphine and β-endorphin analgesia elicited from the amygdala of rats

In addition to brainstem sites of action, analgesia can be elicited following amygdala microinjections of morphine and mu-selective opioid agonists. The present study examined whether opioid analgesia elicited by either morphine or beta-endorphin in the amygdala could be altered by either the general opioid antagonist, naltrexone, the mu-selective antagonist, beta-funaltrexamine (BFNA) or the delta 2 antagonist, naltrindole isothiocyanate (Ntii) in the periaqueductal gray (PAG). Both morphine (2.5-5 micrograms) and beta-endorphin (2.5-5 micrograms) microinjected into either the baso-lateral or central nuclei of the amygdala significantly increased tail-flick latencies and jump thresholds in rats. The increases were far more pronounced on the jump test than on the tail-flick test. Placements dorsal and medial to the amygdala were ineffective. Naltrexone (1-5 micrograms) in the PAG significantly reduced both morphine (tail-flick: 70-75%; jump: 60-81%) and beta-endorphin (tail-flick: 100%; jump: 93%) analgesia elicited from the amygdala, indicating that an opioid synapse in the PAG was integral for the full expression of analgesia elicited from the amygdala by both agonists. Both BFNA (68%) and Ntii (100%) in the PAG significantly reduced morphine, but not beta-endorphin analgesia in the amygdala on the tail-flick test. Ntii in the PAG was more effective in reducing morphine (60%) and beta-endorphin (79%) analgesia in the amygdala on the jump test than BFNA (15-24%). Opioid agonist-induced analgesia in the amygdala was unaffected by opioid antagonists administered into control misplacements in the lateral mesencephalon, and the small hyperalgesia elicited by opioid antagonists in the PAG could not account for the reductions in opioid agonist effects in the amygdala. These data indicate that PAG delta 2, and to a lesser degree, mu opioid receptors are necessary for the full expression of morphine and beta-endorphin analgesia elicited from the amygdala.

Serotonin receptor subtype antagonists in the medial ventral medulla inhibit mesencephalic opiate analgesia

Supraspinal opioid analgesia is mediated in part by connections between the midbrain periaqueductal gray (PAG) and the ventral-medial medulla, including the nucleus raphe magnus (NRM) and nucleus reticularis gigantocellularis (NRGC). A serotonergic synapse appears to participate in this pathway since methysergide microinjected into the NRM-NRGC significantly reduced morphine analgesia elicited from the PAG. The present study evaluated the role of specific serotonin receptor subtypes by pretreating rats with microinjections of either the 5HT2 antagonist, ritanserin or the 5HT3 antagonist, ICS205930, into the NRM-NRGC and examining their effects upon morphine (2.5 micrograms) analgesia elicited from the PAG. Mesencephalic morphine analgesia was significantly reduced following pretreatment with both ritanserin (0.25-2.5 micrograms) on the tail-flick (81%) and jump (65%) tests and ICS205930 (0.25-5 micrograms) on the tail-flick (91%) and jump (63%) tests. Neither ritanserin nor ICS205930 altered basal nociceptive thresholds. Medullary placements ventral or lateral to the NRM/NRGC failed to support these antagonistic effects. These data indicate that ventro-medial medullary 5HT2 and 5HT3 serotonergic receptors modulate the transmission of opioid pain-inhibitory signals from the PAG.

Gender-specific and gonadectomy-specific effects upon swim analgesia: Role of steroid replacement therapy

Both gender-specific and gonadectomy-specific effects have been observed for the analgesic responses following continuous and intermittent cold-water swims (CCWS and ICWS respectively): female rats display significantly less analgesia than males, and gonadectomized rats display significantly less analgesia than sham-operated controls. The present study evaluated the effects of steroid replacement therapy with testosterone propionate (TP: 2 mg/kg, SC) upon CCWS and ICWS analgesia on the tail-flick and jump tests and hypothermia in sham-operated or gonadectomized male and female rats. Thirty days following surgery, rats received either no treatment, a sesame oil vehicle or TP for 14 days prior to, and then during testing. Relative to the no treatment condition, repeated vehicle injections in sham-operated rats eliminated the gender-specific, but did not affect the gonadectomy-specific effects upon CCWS and ICWS analgesia. TP reversed the deficits in CCWS and ICWS analgesia observed in both castrated and ovariectomized rats on both pain tests. TP only potentiated CCWS analgesia in sham-operated males on the tail-flick test. TP potentiated CCWS and ICWS hypothermia in gonadectomized rats and in male sham-operated rats. These data indicate that gonadal steroids play a major modulatory role in the etiology of swim analgesia, and that the observed gender effects are sensitive to possible adaptational variables.

Alterations in deprivation, glucoprivic and sucrose intake following general, mu and kappa opioid antagonists in the hypothalamic paraventricular nucleus of rats

While opioid agonists administered into the hypothalamic paraventricular nucleus increase food intake in rats, naloxone reduces deprivation-induced intake. Ventricular administration of either mu (beta-funaltrexamine) or kappa (nor-binaltorphamine) opioid antagonists reduces spontaneous, deprivation, glucoprivic and palatable intake. The present study assessed whether microinjections of either general, mu or kappa opioid antagonists into the paraventricular nucleus altered either deprivation (24 h) intake, 2-deoxy-D-glucose hyperphagia or sucrose intake in rats. Deprivation intake was significantly reduced by nor-binaltorphamine (5 micrograms, 68 nmol, 30-33%), beta-funaltrexamine (5 micrograms, 100 nmol, 26-29%) or naltrexone (10 micrograms, 260 nmol, 26%) in the paraventricular nucleus. 2-Deoxy-D-glucose hyperphagia was significantly reduced only after 2 h by naltrexone (10 micrograms, 260 nmol, 69%), norbinaltorphamine (20 micrograms, 272 nmol, 69%) or beta-funaltrexamine (20 micrograms, 400 nmol, 83%) in the paraventricular nucleus. Sucrose intake was significantly reduced by nor-binaltorphamine (5 micrograms, 68 nmol, 27-36%), naltrexone (5-10 micrograms, 130-260 nmol, 18-31%) and beta-funaltrexamine (5 micrograms, 100 nmol, 20%) in the paraventricular nucleus. These data indicate that general, mu and kappa opioid antagonists administered into the hypothalamic paraventricular nucleus produce similar patterns of effects upon different forms of food intake as did ventricular administration, implicating this nucleus as part of the circuitry underlying opioid mediation of ingestion.

Inhibition of mesencephalic morphine analgesia by methysergide in the medial ventral medulla of rats

The neural substrates of endogenous supraspinal opioid pain inhibition are mediated in part by connections between the midbrain periaqueductal gray (PAG) and the ventral-medial medulla, including the nucleus raphe magnus (NRM) and nucleus reticularis gigantocellularis (NRGC). To ascertain whether a serotonergic synapse participated in this pathway, the present study determined whether microinjections of methysergide into the NRM or NRGC would alter analgesia elicited by morphine microinjections into the PAG. Morphine (2.5 micrograms) in the PAG and immediately adjacent areas produced significant analgesia on the tail-flick and jump tests in rats. Pretreatment with the serotonin receptor antagonist methysergide (0.5-5 micrograms) in either the NRM or NRGC significantly reduced morphine analgesia elicited from the PAG by 69% on the tail-flick and by 50% on the jump tests without altering basal nociceptive thresholds. Medullary placements ventral or lateral to the NRM/NRGC failed to support this antagonistic effect. These data indicate that a ventro-medial medullary serotonergic synapse participates in the transmission of opioid pain-inhibitory signals from the PAG.