Effect of various ACTH analogs on lordosis behavior in the female rat

The effect of ACTH and various related analogs on lordosis behavior in female rats was compared with that produced by α-MSH. Ovariectomized rats received 2 μg estradiol benzoate on Day 1 and Day 3 either 0.1 or 0.2 mg progesterone. Four hours later the females were placed with sexually experienced male rats and the lordosis quotient (LQ) noted. These particular doses of progesterone were chosen because they were sub-maximal and produced a proportion of both nonreceptive (LQ less than 50%) and receptive (LQ greater than 50%) rats. Treatment with 20 μg α-MSH on Day 2 stimulated lordosis in nonreceptive rats but inhibited lordosis in the receptive rats.Of the other peptides tested only ACTH4–10 was as effective as α-MSH in facilitating and inhibiting lordosis behavior. ACTH1–24 and ACTH4–9 also produced both effects. ACTH1–39 and ACTH1–16, on the other hand, had neither effect but were both effective in stimulating and inhibiting lordosis when administered on Days 1, 2 and 3. It is suggested that ACTH4–10 may contain the essential sequence for these facilitatory and inhibitory effects on female sexual receptivity and that elongation of the peptide chain beyond ACTH 1–13 (α-MSH) may decrease this activity.     

alpha-MSH and Pro-Leu-Gly-NH2 (PLG; MIF-1): influence on dopamine (DA) uptake in crude synaptosomal preparations from rat mediobasal hypothalamus (MBH) and caudate putamen (CP)

The uptake of tritiated dopamine [3H] (DMI insensitive DA uptake) by synaptosomal fractions isolated from rat mediobasal hypothalamus (MBH) and caudate putamen (CP) was measured in the presence of different concentrations of alpha-melanocyte stimulating hormone (alpha-MSH) and Pro-Leu-Gly-NH2 (PLG; MIF-1) which is an inhibitor of alpha-MSH release. Compared to control, [3H]DA uptake increased significantly when the synaptosomal fraction of CP was incubated with 0.1 and 1 microM of alpha-MSH and also when the rat was previously injected with alpha-MSH. A simultaneous reduction of endogenous dopamine content was observed. Kinetic studies suggest that the enhanced uptake induced by alpha-MSH 1 microM is the consequence of the rise in Vmax, without changes in the apparent km. The uptake of [3H]DA in hypothalamic (MBH) preparations on the other hand, was not modified by the presence of alpha-MSH. PLG did not have any significant effect on [3H]DA uptake either in the CP or in the MBH. alpha-MSH may act as a modulator of the dopaminergic nigrostriatal system and the results obtained incubating CP synaptosomes in its presence demonstrate a possible direct modulator action by alpha-MSH on the terminal area of the substantia nigra neurons.     

Influence of centrally administered peptides on ear withdrawal from heat in the rabbit

Certain neuropeptides previously linked to stress and implicated in CNS control of analgesia/algesia were tested using a recently developed analgesiometric model, the rabbit ear-withdrawal test. The latency to ear withdrawal increased in a dose-related manner after β-endorphin was injected intracerebroventricularly (IVC). Intermediate doses (0.5 and 1.0 μg) of adrenocorticotropic hormone (ACTH) caused hyperalgesia as indicated by decreases in latency. Corticotropin-releasing factor (CRF, 0.5 and 1.0 μg) also caused significant hyperalgesia late in the testing period. -Melanocyte stimulating hormone (-MSH, 0.25–2.0 μg), a molecule that shares the first 13 amino acid sequence with ACTH, and somatostatin (0.25–2.0 μg), caused no significant change in latency. However, 1.0 μg doses of each peptide antagonized the analgesic effect of β-endorphin (1.0 μg) in the following order of potency: ACTH = -MSH > CRF > somatostatin. The results support the idea that CNS peptides that are released during stress can exert opposing actions on acute pain, even though they may cause little effect alone.     

Exocytotic release of dopamine in ventral tegmental area slices from C57BL/6 and dopamine transporter knockout mice

The present study used voltammetry to ascertain whether electrically stimulated somatodendritic dopamine release in ventral tegmental area slices from C57BL/6 and dopamine transporter knockout mice was due to exocytosis or dopamine transporter reversal, as has been debated. The maximal concentration of electrically evoked dopamine release was similar between ventral tegmental area slices from dopamine transporter knockout and C57BL/6 mice. Dopamine transporter blockade (10 μM nomifensine) in slices from C57BL/6 mice inhibited dopamine uptake but did not alter peak evoked dopamine release. In addition, dopamine release and uptake kinetics in ventral tegmental area slices from dopamine transporter knockout mice were unaltered by the norepinephrine transporter inhibitor, desipramine (10 μM), or the serotonin transporter inhibitor, fluoxetine (10 μM). Furthermore, maximal dopamine release in ventral tegmental area slices from both C57BL/6 and dopamine transporter knockout mice was significantly decreased in response to Na⁺ channel blockade by 1 μM tetrototoxin, removal of Ca²⁺ from the perfusion media and neuronal vesicular monoamine transporter inhibition by RO-04-1284 (10 μM) or tetrabenazine (10 and 100 μM). Finally, the glutamate receptor antagonists AP-5 (50 and 100 μM) and CNQX (20 and 50 μM) had no effect on peak somatodendritic dopamine release in C57BL/6 mice. Overall, these data suggest that similar mechanisms, consistent with exocytosis, govern electrically evoked dopamine release in ventral tegmental area slices from C57BL/6 and dopamine transporter knockout mice.     

Opposite modulation of capsaicin-evoked substance P release by glutamate receptors

Substance P and glutamate are present in primary afferent C-fibers and play important roles in persistent inflammatory and neuropathic pain. In the present study, we have examined whether activation of different glutamate receptor subtypes modulates the release of substance P evoked by the C-fiber selective stimulant capsaicin (1 μM) from rat trigeminal nucleus slices. The selective NMDA glutamate receptor agonist L-CCG-IV (1–10 μM) enhanced capsaicin-evoked substance P release about 100%. This facilitatory effect was blocked by 0.3 μM MK-801, a selective NMDA receptor antagonist. The metabotropic glutamate receptor agonists L-AP4 (group III) and DHPG (group I) (30–100 μM) inhibited capsaicin-evoked substance P release by approximately 60%. These inhibitory effects were blocked by the selective metabotropic glutamate receptor antagonist (±)-MCPG (5 μM). On the other hand, AMPA and kainate (0.1–10 μM), did not significantly affect capsaicin-evoked substance P release. Thus, substance P release from non-myelinated primary afferents, and possibly nociception, may be under the functional antagonistic control of some metabotropic and ionotropic glutamate receptor subtypes.     

Effect of Pro-Leu-Gly-NH2 on plasma levels of α-MSH in the rat

With the idea to give further support that Pro-Leu-Gly-NH₂ (PLG) acts as MIF (the inhibitor factor of MSH), this paper reports the effect of PLG on the secretion of MSH release using a recently developed radioimmunoassay for α-MSH.Pro-Leu-Gly-NH₂ was effective in inhibiting the release of MSH induced by the injection of acid extracts of median eminence (MRF). The rise in plasma MSH by these extracts was not due to their intrinsic content of MSH. Pro-Leu-Gly-NH₂ also inhibited the basal secretion of MSH in male rats. Since PLG also blocked the release of MSH induced by the injection of haloperidol, it is suggested that its effect is not mediated by dopamine.     

Stimulation by melanocortins of neurite outgrowth from spinal and sensory neurons in vitro

Using ELISAs for B-50/GAP43 and neurofilament (NF), we tested ACTH(1–24), -MSH, ACTH(4–10), and an ACTH(4–9) analogue (ORG2766) for their ability to induce sprouting and neuritogenesis from spinal and sensory neurons. Dissociated fetal rat spinal cord neurons or neonatal rat dorsal root ganglion (DRG) cells were cultured with peptide and assayed after 24, 48, or 96 h. In spinal neurons, -MSH and ACTH(1–24) induced the expression of B-50 dose dependently. After 24 h -MSH had a stimulatory effect (from 10 nM onwards), with a maximum at 100 μM (36% increase). After 96 h the maximal effect of 100 μM -MSH on B-50/GAP43 was lower (19%). ACTH(1–24) (100 μM) stimulated B-50/GAP43 by 19%. Neurofilament levels (96 h) were elevated maximally by 64% at 100 μM -MSH. In DRG neurons a bell-shaped dose-response curve was found for -MSH, the maximal effect being observed after 48 h at 100 nM: 54% for B-50/GAP43 and 22% for NF. In both culture systems neither ACTH(4–10) nor ORG2766 was effective. We conclude that -MSH stimulates the expression of B-50/GAP43 (sprouting) and the formation of NF (neurite elongation) and may therefore be considered a neurotrophic factor.     

Stimulatory effect of prostaglandin E1 on thyroliberin-induced α-melanotropin release from perifused neuro-intermediate lobes of frog pituitary gland

A possible direct effect of prostaglandins on α-melanotropin (α-MSH) release at the level of the intermediate lobe of the frog pituitary was investigated $\text{in vitro}$ using a perifusion system technique. The effect of prostaglandins was studied on both spontaneous and TRH-stimulated α-MSH secretion. No significant effect of PGE₁, PGE₂, PGF_1α or PGF_2α on basal release of α-MSH could be detected. Indomethacin did not alter the α-MSH release induced by TRH. Conversely a significant increase in TRH-induced α-MSH secretion was observed in the presence of 1 x 10^?6M PGE₁. This magnifying effect was directly related to the concentration of TRH for doses ranging from 1 x 10^?8M to 1 x 10^?6M.     

Involvement of hypothalamic somatostatin in glucagon-induced suppression of growth hormone secretion in conscious rats

The role of central glucagon in regulating GH secretion was studied in conscious male rats with chronic indwelling intra-atrial and intracerebro-ventricular (ICV) cannulae. Repeated blood sampling every 20 min from 1000 hr to 1700 hr showed two major GH bursts occurring at regular intervals (3.6±0.1 hr) around 1200 hr and 1540 hr. The ICV (lateral ventricle) injection of glucagon (10 μg/rat) at 1100 hr inhibited spontaneous GH secretion, and the mean (±SE) plasma GH levels from 1120 hr to 1700 hr were lower than those in controls injected ICV with the vehicle solution only (31.9±7.8 ng/ml vs. 157.1±13.4 ng/ml, p<0.01). The GH bursts did not appear until 5 hr after the injection. The intravenous (IV) injection of glucagon (10 μg/rat) did not change plasma GH levels or the occurrence of spontaneous GH bursts. The glucagon-induced suppression of GH release was attenuated when anti-somatostatin serum (ASS), but not normal rabbit serum (NRS), was given IV in a volume of 0.25 ml immediately before the ICV injection of glucagon (10 μg/rat) (mean GH levels at 1120–1700 hr: ASS+glucagon, 133.6±26.7 ng/ml vs. NRS+glucagon, 30.5±7.4 ng/ml, p<0.01). These findings suggest that central glucagon may play an inhibitory role in regulating GH secretion by stimulating SRIF release from the hypothalamus in the rat.     

The in vivo modulation of dopamine synthesis by calcium ions: influences on the calcium independent release

To investigate the contribution of the dopamine (DA) synthesis to both the calcium-dependent and the carrier-mediated, mechanisms of DA release in the striatum, anaesthetized rats were locally superfused in the striatum with a push–pull cannula supplied with an artificial CSF containing tritiated tyrosine. DA, dihydroxyphenylacetic acid (DOPAC) and their respective specific activity were measured in effluent and used to evaluate changes in the DA synthesizing rate. Excluding calcium ions from the CSF only partially reduced spontaneous DA release (70%) still leaving a possible carrier-mediated DA release. This effect was not additive with a local superfusion with 0.1 mM a-methyl-p-tyrosine, a blocker of DA synthesis, suggesting that synthesis could already be reduced by calcium-free superfusion. Local superfusion with 100 μM cadmium in the presence or not of calcium ions, increased the DA release (220 and 350%, respectively), simultaneously reducing DA synthesis. Local application of 1 μM calcium ionophore (A23187) was without effect on the basal release of DA but enhanced DA synthesis and increased the amphetamine-evoked and carrier-mediated amine release. We conclude that DA synthesis can be a modulatory process of the firing-independent and carrier-mediated amine release while it weakly affects the classical calcium-dependent release.     

Dopaminergic Inhibition of Prolactin Release and Calcium Influx Induced by Neurotensin in Anterior Pituitary Is Independent of Cyclic AMP System

The present study demonstrates that 3,4-dihydroxyphenylethylamine (DA, dopamine) prevents neurotensin (NT) stimulation of both prolactin (PRL) release and calcium influx by interacting with specific receptors that are functionally linked to calcium channels. As shown by the studies with dispersed cells from rat anterior pituitary, the pharmacology of the control of PRL release and calcium influx, both induced by NT, was found to be typical of a DAergic process. This was demonstrated by the order of potency of agonists in inhibiting PRL release and calcium influx (DA greater than epinephrine greater than norepinephrine much greater than isoproterenol); by the high affinity of antagonists such as haloperidol and fluphenazine for this process; and by the high degree of stereoselectivity of sulpiride. Specific D2 receptor agonists, such as bromocriptine and lisuride, and the specific D2 receptor antagonist (-)-sulpiride were found to be highly potent on the DA receptors negatively coupled with calcium channels and PRL release. DA was found to lack the capacity to change the influx of calcium induced by either the sodium channel activator veratridine or high extracellular potassium levels, thus indicating a specific action of this amine on calcium channels sensitive to NT. In a range of concentrations that are effective in inhibiting either the calcium influx or the PRL release, both induced by NT, DA did not alter the cyclic AMP generating system. DA (from 1.0 nM to 50 nM) did not affect adenylate cyclase activity in rat pituitary gland homogenates and did not modify intracellular cyclic AMP levels in pituitary cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

μ-opioid receptor-mediated inhibition of the release of radiolabelled noradrenaline and acetylcholine from rat amygdala slices

Presynaptic modulation by opioids of electrically-evoked neurotransmitter release from superfused rat amygdala slices prelabelled with [³H]noradrenaline (NA) and [¹⁴C]choline was examined. Both [³H]NA and [¹⁴C]acetylcholine release were strongly inhibited by morphine, the mixed δ/μ-receptor agonist [ -Ala², -Leu⁵]enkephalin (DADLE) and the highly selective μ-agonist [ -Ala², MePhe⁴, Gly-ol⁵]enkephalin (DAMGO), whereas the highly selective δ-agonist [ -Pen², -Pen⁵]enkephalin and the κ-agonist bremazocine were without effect. The inhibitory effects were potently antagonized by naloxone but not by the selective δ-receptor antagonist fentanylisothiocyanate. When the selective uptake inhibitor desipramine was used to prevent uptake of [³H]NA into noradrenergic nerve terminals, but sparing the uptake into dopaminergic nerve terminals, the electrically evoked release of tritium was strongly inhibited by bremazocine but not by DADLE or DAMGO.

The data indicate, that in the amygdala transmitter release from dopaminergic nerve fibres is inhibited only via activation of κ-receptors, whereas transmitter release from noradrenergic and cholinergic nerve fibers is subjected to inhibition by opioids via activation of μ-receptors only. Regional differences and similarities of modulation of neurotransmitter release by opioids in the rat brain are briefly discussed.     

Effect of Nitroprusside (Nitric Oxide) on Endogenous Dopamine Release from Rat Striatal Slices

It is becoming apparent that the synthesis of nitric oxide (NO) from L-arginine not only explains endothelium-dependent vascular relaxation, but is a widespread mechanism for the regulation of cell function and communication. We examined the role of NO on the endogenous dopamine (DA) release from rat striatum. Nitroprusside, in the concentration range of 3-100 microM, induced a dose-dependent increase in the endogenous DA release from rat striatal slices. The maximal response was 330% over the baseline release. A higher concentration of nitroprusside (300 microM) produced an inhibitory effect on the spontaneous release of DA. L-Arginine (10 and 100 microM), a substrate in the NO-forming enzyme system, also produced an elevation of DA release. L-Arginine-induced DA release was attenuated by NG-monomethyl-L-arginine, an inhibitor of NO synthase. NADPH (1 microM), a cofactor of NO synthase, enhanced L-arginine-induced DA release. These results suggest a possible involvement of NO in the DA release process in rat striatum.     

Age-dependent modulation by galanin of growth hormone release from rat pituitary cells in culture.

The effect of galanin (Gal), a 29-amino acid peptide widely distributed in mammalian CNS, was investigated in cultured pituitary cells of rats of different ages. Gal (0.1-10/microM) stimulated GH release in a concentration-dependent manner in 5-and 10-day-old rat pituitaries (EC50: 0.87 and 1.73/microM, respectively) but was ineffective (0.01-1/microM) or even inhibitory (10/microM) in 40-day-old male rat pituitaries. Gal (0.1-10/microM) was ineffective to alter stimulation of GH release induced by GHRH (10 nM) in 5-day-and 40-day-old rat pituitary cells, but Gal (1/microM) slightly inhibited (24%) it in 10-day-old rat pituitaries. Gal (1 and 10/microM) also inhibited GH secretion (45 and 58%, respectively) from 40-day-old pituitary cells when a lower GHRH dose (0.1 nM) was used for stimulation. The results of this study indicate that Gal has the ability to either stimulate or inhibit GH release from dispersed pituitary cells and that its effects are closely related to the age of the rats.     

Arachidonate lipoxygenase inhibitors in guinea-pig isolated trachea. Effects on contractions to antigen and various agonists

We compared the effects of the leukotriene (LT) D₄ receptor antagonist FPL55712 and some lipoxygenase inhibitors on contractions of isolated guinea-pig trachea induced by antigen (ovalbumin, OA) and calcium ionophore A23187 in the presence of the cyclooxygenase inhibitor indomethacin (5 μM), and by arachidonic acid (AA), melittin and LTD₄. FPL55712 (0.1 and 1 μM) inhibited contractions induced by AA (100 μM) and the phospholipase A₂ activator melittin (3 μg/ml), while the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA, 10 μM) was a more effective inhibitor of the melittin response than the response. FPL55712 inhibited contractions induced by OA (100 μg/ml) more than by A23187 (1 μg/ml), and these inhibitory effects of FPL55712 were much less in the presence of l-serine-borate complex (45 mM), an inhibitor of LTC₄ conversion to LTD₄. NDGA (10 μM) had no significant effect on the OA response, whereas the lipoxygenase inhibitors 1-phenyl-3-pyrazolidone (phenidone, 10 μM) and 5,8,11,14-eicosatetraynoic acid (ETYA, 10 μM) clearly inhibited it. In contrast, NDGA and phenidone inhibited the A23187 response, but ETYA had no effect on it. FPL55712, phenidone and ETYA, but not NDGA, had a large inhibitory effect on LTD₄-induced contractions, but these inhibitors had no effect on histamine-induced contractions. These results suggest that in the guinea-pig trachea inhibitors of LTD₄-induced contractions decrease antigen-induced contractions, whereas lipoxygenase inhibitors reduce the contraction to A23187.     

Ethanol-Induced Increase in Endogenous Dopamine Release May Involve Endogenous Opiates

The effect of opiate peptides on basal and potassium-stimulated endogenous dopamine (DA) release from striatal slices was studied in vitro. Dual stimulation of the striatal slices gave a reproducible increase in DA release that was calcium dependent. Addition of the delta-opiate receptor agonists Met5-enkephalin, [D-Ala2,D-Leu5]enkephalin (DADLE), and [D-Ser2]Leu-enkephalin-Thr (DSLET), increased the basal DA release without affecting potassium-stimulated release in a dose-dependent manner. The effect of DADLE was antagonized by the addition of naloxone. In contrast, the mu-opioid receptor agonist [D-Ala2,N-MePhe4,Gly-ol5]enkephalin (DAGO) and the epsilon-opioid agonist beta-endorphin inhibited the stimulated DA release without changing the basal release. The inhibitory effect of DAGO on potassium-stimulated release was antagonized by naloxone. The addition of ethanol (75 mM) to the incubation media produced a delayed increase of both the basal and stimulated DA release. There was no change in stimulated DA release when the change in basal release was subtracted, suggesting that ethanol produced a dose-dependent, selective increase in basal DA release. Naloxone and the selective delta-opiate antagonist ICI 174864 inhibited the ethanol-induced increase in basal DA release. Naloxone and ICI 174864 added alone did not alter either basal or stimulated DA release. We therefore suggest that the ethanol-induced increase in basal DA release is an indirect effect involving an endogenous delta-opiate agonist.     

Potent inhibitory effect of selective D2 and D3 agonists on dopamine-responsive dorsomedial arcuate neurons in brain slices of estrogen-primed rats.

The possible involvement of dopamine D2 and D3 receptors in the action of dopamine (DA) on inhibiting dorsomedial arcuate nucleus (dmARN) neurons in brain slices was determined in this study. Fresh brain slices were prepared from ovariectomized, estrogen-primed Sprague-Dawley rats and used for extracellular single-unit recording. The dmARN neurons were first identified by their inhibitory responses to DA and then tested with PHNO and/or PD128907, selective D2 and D3 agonists, respectively. PD128907 in 5-50 nmole doses significantly inhibited the majority of DA-responsive dmARN neurons (86.3% of 44 units). Moreover, PHNO in 5-25 nmole doses inhibited all DA-responsive neurons tested (100% of 34 units). The inhibitory effects of PHNO and PD128907 were not only prominent; but also persisted in low Ca2+, high Mg2+ medium, indicating that they were acting directly on the recorded neuron. Pretreatment of either raclopride or U99194A, D2 and D3 receptor antagonists respectively, reversed the effects of DA in a few trials. In contrast, SKF81297, a D1 receptor agonist, induced variable responses in dmARN neurons. These results clearly indicate that DA may act through D2 and/or D3 receptors to exhibit an inhibitory effect on presumed TIDA neurons in dmARN.     

几种神经内分泌因子对鲤促性腺激素释放激素(GnRH)释放的调节作用:离体研究

用离体静态培育系统进行的初步研究表明 ,在幼鲤 ,多巴胺 (DA)显著刺激下丘脑片段和脑垂体碎片释放GnRH ,并且是剂量依存的 ;促甲状腺素释放激素 (TRH)和γ -氨基酸丁酸 (GABA)对GnRH的释放没有影响。在成鲤 ,DA抑制下丘脑片段和脑垂体碎片释放GnRH ,而TRH和GABA刺激GnRH的释放 ;DA对GABA刺激的GnRH释放也具有抑制作用 ;TRH和GABA的协同作用对下丘脑和脑垂体GnRH释放活动的影响明显低于TRH和GABA的单独作用 ,表明TRH和GABA之间可能存在着某种GnRH释放的相互消竭作用。