Further characterization of 5-HT1A receptors in the goldfish retina: role of cyclic AMP in the regulation of the in vitro outgrowth of retinal explants.

The presence of serotonin 5-HT1A receptors and their physiological role were further characterized in the goldfish retina. The effects of the 5-HT6/7 receptor antagonists pimozide, fluphenazine and amoxapine, the 5-HT1A receptor antagonist WAY-100,135, and the alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, on the 5-HT1A receptor agonist [3H]8-hydroxy-2-(di-n-propylamino)tetralin binding to retinal membranes, were evaluated. In addition, the effects of serotonin, 8-hydroxy-2-(di-n-propylamino)tetralin, WAY-100,135, the adenylate cyclase inhibitors SQ22536 and MDL12330A, and the cyclic AMP analog 8-bromoadenosine-3':5' cyclic monophosphate were also studied on neuritic outgrowth from retinal explants. WAY-100,135 but not 5-HT6/7 receptor antagonists inhibited [3H]8-hydroxy-2-(di-n-propylamino)tetralin binding to retinal membranes N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline decreased [3H]8-hydroxy-2-(di-n-propylamino)tetralin binding sites up to 70%, while receptor turnover was similar to that reported in other tissues. Serotonin and 8-hydroxy-2-(di-n-propylamino)tetralin stimulated cyclic AMP production, both ex vivo and in vitro, and these increases were related to inhibition of neuritic outgrowth. The inhibitory effect was reduced by SQ22536 and by WAY-100,135, and was mimicked by 8-bromoadenosine-3':5'cyclic monophosphate. This study supports previous findings about the role of serotonin as a regulator of axonal outgrowth during in vitro regeneration of the goldfish retina and demonstrates that this effect is mediated, at least in part, by 5-HT1A receptors through a mechanism which involves an increase of cyclic AMP levels.

[Collaboration between the geriatrician and the surgeon: a major contribution for the aged patient in rehabilitation]. / La collaboration entre le gériatre et le chirurgien: un intérêt majeur pour le patient âgé en rééducation.

BACKGROUND: Subtrochanteric fractures are complex fractures requiring prudent reeducation. In elderly patients who also have other illnesses, reeducation of walking may be quite difficult. CASE REPORT: An 89-year-old woman was referred for physical therapy after osteosynthesis of a comminutive subtrochanteric fracture. Weight bearing was contraindicated for 3 months due to her "incapacity to perform reeducation exercises". The patient also had several serious co-morbidities which had to be integrated into the reeducation program. DISCUSSION: In cases of complex fractures, reeducation must take into account other comorbid conditions. Outcome can be favorable, but requires a well organized interdisciplinary approach associating the geriatrics and the orthopedics teams.

Modulation of outgrowth from goldfish retinal explants by a 5-HT2 receptor agonist and [3H]ketanserin binding sites in goldfish and rabbit retina.

The binding of [3H]ketanserin to goldfish and rabbit retinal membrane preparations and the possible role of 5-HT2A receptors in the in vitro outgrowth from goldfish retina were evaluated. Saturation experiments indicated a high-affinity binding site, and positive cooperativity for both tissues. The 5-HT2A/2C agonist (+/-)-2,5-dimetoxy-4-iodoamphetamine and serotonin inhibited outgrowth from goldfish retinal explants. These effects were blocked by the 5-HT2 antagonists ketanserin and 1-(1-naphthyl)piperazine and by the 5-HT2C antagonist mesulergine, respectively. Results make to suggest that [3H]ketanserin binds to 5-HT2A receptors in the rabbit and goldfish retina, but also to a monoamine transporter in the latter tissue. Subtypes of 5-HT2 receptors might mediate the 5-HT modulatory role on in vitro outgrowth of the goldfish retina.

5HT1A receptor agonist differentially increases cyclic AMP concentration in intact and lesioned goldfish retina. In vitro inhibition of outgrowth by forskolin.

5HT1A receptors occur in the retina of various species and the administration of 5HT1A agonists results in the inhibition of outgrowth from postcrush goldfish retinal explants. The levels of cyclic AMP (cAMP) play a role in the modulation of the outgrowth of the nevous system. Moreover, the stimulation of central 5HT1A receptors with the agonist 8-hydroxy-2-(di-n-propylamino)tetralin has been reported to produce an increase or decrease in the activity of adenylate cyclase. In the present investigation we studied the effect of adenylate cyclase stimulation by forskolin, as well as the modulatory effects of 5HT1A receptor agonists and antagonists on the production of cAMP in the goldfish retina, and on the outgrowth of this tissue in vitro. 8-Hydroxy-2-(di-n-propylamino)tetralin produced a significant and dose-dependent increase in cAMP concentration. This effect was not additive to the stimulation produced by forskolin. By contrast, as previously described, the 5HT1A agonist decreased cAMP concentration in the hippocampus of the rat. Both effects were significantly impaired by the 5HT1A antagonist WAY-100,135. A significant effect of the antagonist alone was observed only in the goldfish retina. The increase in cAMP levels was greater in the intact than in the postcrush retina. In addition, forskolin decreased the outgrowth of postcrush retinal explants in a dose-dependent manner, suggesting the importance of critical levels of cAMP in this process. Taken together, 5HT1A receptors seem to be positively coupled to adenylate cyclase in the goldfish retina, where cAMP plays a role as a modulator of outgrowth and regeneration. The inhibitory effect of 5HT1A receptor agonists on retinal outgrowth might be mediated through the production of cAMP. The activation of other subtypes of 5HT receptors positively coupled to adenylate cyclase by the 5HT1A agonist, such as 5HT7, cannot be discarded.

8-[3H]hydroxy-2-(di-n-propylamino) tetralin binding sites in goldfish retina.

The binding sites of 8-[3H]hydroxy-2-(di-n-propylamino)tetralin ([3H]DPAT) were characterized in the retina of goldfish in order to evaluate the selectivity of the ligand for serotonin1A (5HT1A) receptors. Specificity of the binding was performed in the presence of serotonergic and dopaminergic agonists and antagonists. Buspirone, spiroxatrine and 5-methoxy-N,N-dimethyltryptamine were potent inhibitors, followed by propranolol, citalopram, imipramine and desipramine. Serotonin was not a potent inhibitor, and its interaction with the binding sites of [3H]DPAT was complex. Nomifensine displayed an important inhibition, however, other dopamine uptake blockers, such as bupropion and GBR-12909, were less potent. Haloperidol was also a good inhibitor, but the D1 receptor agonist, SKF-38393, the D2 receptor antagonist, sulpiride, and dopamine did not inhibit the binding. GppNHp inhibited the binding in the micromolar range. The analysis of saturation experiments by isotopic dilution, using buspirone to determine nonspecific binding, revealed two sites. The number of binding sites defined by buspirone were higher than the ones defined by nomifensine. The specific binding, using buspirone for definition, was reduced by the intraocular injection of 6-hydroxydopamine. This investigation demonstrates that [3H]DPAT labels 5HT1A receptors in goldfish retina, but also interacts with a non-5HT receptor site. These receptors seem to be localized in dopaminergic neurons.

Characterization of serotonin transporter in goldfish retina by the binding of [3H]paroxetine and the uptake of [3H]serotonin: modulation by light.

The serotonin (5-HT) uptake system of goldfish retina was evaluated by the binding of [3H]paroxetine to membrane preparations and the uptake of [3H]5-HT into isolated cells from goldfish retina. The order of potency of inhibitors of [3H]paroxetine binding was imipramine > 5-methoxy-N,N- dimethyltryptamine > desipramine > fluoxetine > citalopram > 5-HT. The saturation experiments indicated a high-affinity binding site, and positive cooperativity with Hill coefficient higher than unity. The association reached equilibrium at about one hour of incubation and was efficiently displaced by imipramine. The equilibrium dissociation constants calculated by the antilog of the log concentration of ligand giving 50% of occupation, and by the ratio of dissociation/association constants, were similar: 5.84 and 2.34 nM, respectively. The binding was not significantly reduced by decreasing the temperature of incubation and was sodium dependent. The lesion with 5,7-dihydroxytryptamine reduced the binding to 60%. The uptake of [3H]5-HT into isolated cells also showed positive cooperativity. The order of potency of inhibitors was similar to the one obtained for the binding of [3H]paroxetine. Darkness increased the uptake of 5-HT. The allosteric regulation of the 5-HT transporter and the modulation by light could be related to the physiological role of the monoamine, as a neurotransmitter and as a precursor of melatonin synthesis in the retina.