RESUMEN
Dental enamel forms through the concerted activities of specialized extracellular matrix proteins, including amelogenin, enamelin, MMP20, and KLK4. Defects in the genes encoding these proteins cause non-syndromic inherited enamel malformations collectively designated as amelogenesis imperfecta (AI). These genes, however, account for only about a quarter of all AI cases. Recently we identified mutations in FAM83H that caused autosomal dominant hypocalcified amelogenesis imperfecta (ADHCAI). Unlike other genes that cause AI, FAM83 H does not encode an extracellular matrix protein. Its location inside the cell is completely unknown, as is its function. We here report novel FAM83H mutations in four kindreds with ADHCAI. All are nonsense mutations in the last exon (c.1243G>T, p.E415X; c.891T>A, p.Y297X; c.1380G>A, p.W460X; and c.2029C>T, p.Q677X). These mutations delete between 503 and 883 amino acids from the C-terminus of a protein normally comprised of 1179 residues. The reason these mutations cause such extreme defects in the enamel layer without affecting other parts of the body is not known yet. However it seems evident that the large C-terminal part of the protein is essential for proper enamel calcification.
Asunto(s)
Análisis Mutacional de ADN , Proteínas/química , Proteínas/genética , Calcificación de Dientes/genética , Adolescente , Amelogénesis Imperfecta/genética , Niño , Exones , Matriz Extracelular/metabolismo , Femenino , Genes Dominantes , Humanos , Masculino , Modelos Genéticos , Fenotipo , Estructura Terciaria de Proteína , Proteínas/fisiologíaRESUMEN
Nonsyndromic dentin defects classified as type II dentin dysplasia and types II and III dentinogenesis imperfecta are caused by mutations in DSPP (dentin sialophosphoprotein). Most reported disease-causing DSPP mutations occur within the repetitive DPP (dentin phosphoprotein) coding sequence. We characterized the DPP sequences of five probands with inherited dentin defects using single molecule real-time (SMRT) DNA sequencing. Eight of the 10 sequences matched previously reported DPP length haplotypes and two were novel. Alignment with known DPP sequences showed 32 indels arranged in 36 different patterns. Sixteen of the 32 indels were not represented in more than one haplotype. The 25 haplotypes with confirmed indels were aligned to generate a tree that describes how the length variations might have evolved. Some indels were independently generated in multiple lines. A previously reported disease-causing DSPP mutation in Family 1 was confirmed and its position clarified (c.3135delC; p.Ser1045Argfs*269). A novel frameshift mutation (c.3504_3508dup; p.Asp1170Alafs*146) caused the dentin defects in Family 2. A COL1A2 (c.2027G>A or p.Gly676Asp) missense mutation, discovered by whole-exome sequencing, caused the dentin defects in Family 3. We conclude that SMRT sequencing characterizes the DPP repeat region without cloning and can improve our understanding of normal and pathological length variations in DSPP alleles.
RESUMEN
Dose-responsive motor activity induced by systemic injection of the kappa (kappa) preferring opioid agonist, U50,488H (1-10 mg/kg, s.c.) in guinea pigs was recently reported [Brent P. J. and Bot G. (1992) Psychopharmacology 107: 581-590], characterised at the higher doses used (5-10 mg/kg) by sustained postural abnormalities. The effects on the U50,488H-induced, abnormal, motor response of pharmacological manipulation of opioid receptors and sigma (sigma) sites was studied. The opioid antagonist naloxone, [5 and 15 mg/kg, subcutaneously (s.c.)], the kappa selective antagonist, norbinaltorphimine (NBNI), administered intracerebroventricularly (i.c.v., 20 and 50 nM) 0.5 hr before U50,488H, and the anticonvulsant phenytoin [25 and 50 mg/kg, intraperitoneally (i.p.)] given 1 hr before, attenuated the abnormal postures, whereas naloxone methobromide (15 mg/kg), a quaternary opioid which does not cross the blood-brain barrier, had no significant effect on the movements. In contrast, the drugs with varying affinity for sigma binding sites such as 1,3-di(2-tolyl)guanidine (DTG, 10 and 30 mg/kg), haloperidol (1 and 5 mg/kg, s.c.), dextromethorphan (1, 10 and 20 mg/kg, s.c.) and reduced haloperidol (1 mg/kg, s.c.), given 0.5-1 hr before U50,488H, exacerbated the severity of the abnormal motor activity in a dose-related manner by decreasing the latency to onset of maximum obtainable motor response and increasing the duration of the response. In addition, haloperidol (1 and 5 mg/kg, s.c.), dextromethorphan (10 mg/kg, s.c.) and DTG (30 mg/kg, s.c.), given in combination with U50,488H, induced behaviour characterised by marked oral activity. In contrast to the effect of haloperidol, pretreatment with the selective dopamine D-2 antagonist, raclopride (10 mg/kg, s.c.), had no significant effect on the abnormal movements induced by U50,488H, but did induce oral activity. These data indicate the possible involvement of kappa opioid receptors in the abnormal movement induced by U50,488H, and further demonstrate that there is an interaction between the kappa receptors and sigma sites which can influence the abnormal motor activity.
Asunto(s)
Analgésicos/farmacología , Conducta Animal/efectos de los fármacos , Locomoción/efectos de los fármacos , Antagonistas de Narcóticos/farmacología , Pirrolidinas/farmacología , Receptores Opioides kappa/efectos de los fármacos , Receptores sigma/efectos de los fármacos , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Antagonistas de los Receptores de Dopamina D2 , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Femenino , Cobayas , Inyecciones Intraventriculares , Ligandos , Masculino , Actividad Motora/efectos de los fármacos , Fenitoína/farmacología , Postura , Racloprida , Receptores de Dopamina D2/metabolismo , Salicilamidas/farmacologíaRESUMEN
The effects on locomotor activity and plasma catecholamines of substance P, 0.5 nmol, injected into each lateral ventricle (i.c.v.), or 1 nmol, injected into the cisterna magna of conscious guinea-pigs, were investigated. Locomotor activity was measured in cages fitted with an infra-red photocell and detector, and plasma catecholamines, were measured by HPLC with electrochemical detection. Substance P, given intraventricularly or into the cisterna magna, produced increased locomotor activity and a pattern of behavioural activity which mimicked the opiate withdrawal response, found in previous studies in this species. Levels of NA and AD in plasma were also significantly elevated after injection of substance P. These effects of substance P were relatively long-lasting, since they were present up to 1 hr after injection. The results show that the effects of centrally administered substance P in guinea-pigs are similar to those in rats. Furthermore, the results support the suggestion that substance P might be the mediator of the opiate withdrawal response in the central nervous system as has been proposed for the enteric nervous system.
Asunto(s)
Epinefrina/sangre , Actividad Motora/efectos de los fármacos , Norepinefrina/sangre , Sustancia P/farmacología , Animales , Cisterna Magna , Femenino , Cobayas , Inyecciones , MasculinoRESUMEN
The effects of intracerebroventricular administration of morphine, the selective mu-agonist DAMGO, the delta-agonist DPDPE, the kappa-preferring peptide dynorphin A(1-13) and the kappa-agonist U50,488H on locomotor behaviour in the guinea pig were investigated. Morphine (total dose = 0.01, 0.1, 1, 10, 200 nmol), DAMGO and DPDPE (total dose = 0.1, 1, 10, 100 nmol of each) produced piloerection and sedation, indicating that the responses of guinea pigs to mu- and delta-opioid agonists differed from those of rats and mice. In contrast, U50,488H (total dose = 10, 100 nmol) and dynorphin A(1-13) (total dose = 100 nmol) produced increased locomotor activity which was attenuated by pretreatment with naloxone and norbinaltorphimine, thus confirming the involvement of kappa-opioid receptors. Furthermore, pretreatment with spantide, baclofen, muscimol, bicuculline, MK-801, raclopride and atropine also inhibited the U50,488H-induced locomotor activity, suggesting the involvement of GABA, dopamine, excitatory amino acids, substance P and acetylcholine in this response.
Asunto(s)
Analgésicos/farmacología , Actividad Motora/efectos de los fármacos , Narcóticos/farmacología , Pirrolidinas/farmacología , Receptores Opioides/efectos de los fármacos , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Dinorfinas/farmacología , Encefalina Ala(2)-MeFe(4)-Gli(5) , Encefalina D-Penicilamina (2,5) , Encefalinas/farmacología , Femenino , Cobayas , Hipnóticos y Sedantes/farmacología , Inyecciones Intraventriculares , Inyecciones Subcutáneas , Masculino , Morfina/farmacología , Fragmentos de Péptidos/farmacología , Piloerección/efectos de los fármacos , Receptores Opioides/fisiología , Receptores Opioides delta/efectos de los fármacos , Receptores Opioides delta/fisiología , Receptores Opioides kappa/efectos de los fármacos , Receptores Opioides kappa/fisiología , Receptores Opioides mu/efectos de los fármacos , Receptores Opioides mu/fisiologíaRESUMEN
We present data for a patient, GY, with a right hemianopia caused by traumatic damage to the left occipital cortex. Previous studies have established that this patient has residual vision which enables him to detect and localize transient stimuli presented to his 'blind' hemifield. We have now examined spectral responses associated with this residual vision by using two-colour incremental threshold methods to measure II-spectral functions, and a white light background to examine spectral data for 'colour-opponent' characteristics. We report that both methods yield normal spectral response characteristics for GY's 'blind' hemifield. We have also investigated the patient's ability to identify, verbally, coloured stimuli presented to his 'blind' hemifield, and found that, in 'forced choice' experiments, he achieves a high proportion of correct responses to large stimuli. The patient reported that in threshold detection measurements his responses were based on the presence or absence of a percept associated with transient light stimulation of the 'blind' hemifield (residual vision), whereas colour naming was achieved without conscious perception of colour ('blindsight').
Asunto(s)
Daño Encefálico Crónico/fisiopatología , Percepción de Color , Lóbulo Occipital/lesiones , Accidentes de Tránsito , Adulto , Humanos , Masculino , Estimulación Luminosa , Umbral Sensorial , Trastornos de la Visión/etiología , Trastornos de la Visión/fisiopatologíaRESUMEN
The present study examined the behavioural effects of sigma agonists and PCP-like non-competitive N-methyl-D-aspartate (NMDA) antagonists in guinea-pigs. Subcutaneous (SC) injection of the putative sigma agonist (+)NANM (1 and 10 mg/kg SC) and (-)NANM (1 and 10 mg/kg SC) produced a behavioural response in guinea-pigs which was characterized by sedation and exophthalmos, with locomotor depression, flattened posture and flaccidity, whereas the sigma ligand pentazocine induced sedation but no flattened posture. Ketamine (20 mg/kg SC) and (+)dizocilpine (0.025, 0.1 and 1 mg/kg SC) produced similar effects to those of (+) and (-)NANM. However, the putative sigma receptor ligand DTG (1 and 10 mg/kg SC) had no observable effect on behaviours in guinea-pigs, similar to results for other species. The behavioural effects produced by (+) and (-)NANM were not reversed by injection 1 h later of naloxone hydrochloride (15 mg/kg SC), haloperidol (10 mg/kg SC) or DTG (10 and 30 mg/kg SC), but the effects of all drugs were reversed by the selective dopamine D-2 agonist quinpirole (3 mg/kg IP). Moreover, injection of naloxone (15 mg/kg SC), DTG (10 and 30 mg/kg SC) or haloperidol (1 and 10 mg/kg SC) 10 min before, did not reverse the behaviour induced by (+)NANM (10 mg/kg SC). These data indicate that sigma and PCP-like drugs have a similar gross behavioural effect in guinea-pigs, possibly mediated by non-competitive antagonism of the NMDA subtype of glutamate receptors. The results demonstrating behavioural depression were in contrast to the stimulatory effects of these drugs at similar doses in other rodent species.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Conducta Animal/efectos de los fármacos , N-Metilaspartato/antagonistas & inhibidores , Fenciclidina/análogos & derivados , Fenciclidina/farmacología , Receptores Opioides/efectos de los fármacos , Animales , Antipsicóticos/farmacología , Benzamidas/farmacología , Maleato de Dizocilpina/farmacología , Dopaminérgicos/farmacología , Antagonistas de Dopamina , Ergolinas/farmacología , Femenino , Guanidinas/farmacología , Cobayas , Haloperidol/farmacología , Ketamina/farmacología , Masculino , Actividad Motora/efectos de los fármacos , Pentazocina/farmacología , Fenazocina/análogos & derivados , Fenazocina/farmacología , Quinpirol , Receptores de Dopamina D2 , Receptores sigma , RemoxipridaRESUMEN
This study describes specific behaviours in guinea-pigs after dopamine D-1 and D-2 receptor activation which differed to those described in other rodent species. Intraperitoneal (IP) administration of the dopamine D-2 receptor agonist quinpirole (1.5, 3 and 6 mg/kg) to guinea-pigs dose-dependently initiated locomotor activity and other behaviours including rearing, head-down sniffing, chewing, circling, licking and head/body shaking. Locomotor activity induced by quinpirole (3 mg/kg) was reduced by the D-2 receptor antagonists sulpiride, (100 mg/kg IP) and raclopride (10 mg/kg IP). The dopamine D-1 receptor agonist SKF 38393 (8, 16 and 32 mg/kg IP) produced little or no behavioural effect, nor did the D-1 receptor antagonist SCH 23390 (0.2 and 0.4 mg/kg IP). A 16 mg/kg dose of SKF 38393, given in combination with 3 mg/kg quinpirole, produced responses similar to quinpirole alone, whereas 32 mg kg SKF 38393 combined with quinpirole induced vacuous oral chewing, with attenuation of locomotor activity and circling, but not other behaviours produced by this dose of quinpirole. In contrast to previous studies in rats, the responses to quinpirole (3 mg/kg) were not significantly affected by SCH 23390 (0.2 and 0.4 mg/kg). It is concluded that the guinea-pig may be a useful and interesting species for study of the behavioural effects of D-1 and D-2 agonists and antagonists, as its responses appear to differ from those of other rodent species, but are similar to some responses to D-1 agonists observed in primates.
Asunto(s)
Conducta Animal/efectos de los fármacos , Receptores Dopaminérgicos/efectos de los fármacos , 2,3,4,5-Tetrahidro-7,8-dihidroxi-1-fenil-1H-3-benzazepina/farmacología , Animales , Antagonistas de Dopamina , Relación Dosis-Respuesta a Droga , Ergolinas/farmacología , Femenino , Cobayas , Masculino , Actividad Motora/efectos de los fármacos , Quinpirol , Racloprida , Receptores de Dopamina D1 , Receptores de Dopamina D2 , Salicilamidas/farmacología , Sulpirida/farmacologíaRESUMEN
The effects of morphine D-Pen2, D-Pen5 enkephalin (DPDPE) and U50,488H on the behavioural syndrome elicited by the dopamine (DA) D-2 agonist quinpirole, were investigated. Morphine (1, 5 and 15 mg/kg SC) and morphine administered intracerebroventricularly (ICV) (2 x 5 microliters, 10(-3) M; total dose = 10 nmol) produced piloerection and sedation. DPDPE-ICV (2 x 5 microliters and 2 x 10 microliters, 10(-3) M; total doses = 10 and 20 nmol) produced piloerection and sedation similar to morphine. U50,488H (1 mg/kg SC) induced locomotor activity and some stereotyped behaviour, whereas U50,488H (5 and 10 mg/kg SC) induced muscle rigidity and dystonic-like movements. The locomotor and behavioural response elicited by quinpirole (3 mg/kg IP) was attenuated in guinea-pigs pretreated with morphine (1, 5 and 15 mg/kg SC), morphine-ICV (2 x 5 microliters, 10(-3) M), and DPDPE-ICV (2 x 5 microliters and 2 x 10 microliters, 10(-3) M). These effects were reversed by naloxone (15 mg/kg SC). U50,488H (1 mg/kg SC) increased the quinpirole-induced locomotor activity, whereas U50,488H (5 and 10 mg/kg SC) decreased the locomotor activity and stereotyped behaviours produced by quinpirole. These results indicate that the gross behavioural effects of mu, delta and kappa opioids differ in guinea-pigs compared to other rodent species, and suggest differential involvement of these opioid receptor subtypes with DA D-2 receptor-mediated activity.
Asunto(s)
Conducta Animal/efectos de los fármacos , Encefalinas/farmacología , Ergolinas/farmacología , Morfina/farmacología , Actividad Motora/efectos de los fármacos , Pirrolidinas/farmacología , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Encefalina D-Penicilamina (2,5) , Femenino , Cobayas , Hipnóticos y Sedantes/farmacología , Inyecciones Intraventriculares , Masculino , Piloerección/efectos de los fármacos , QuinpirolRESUMEN
A single dose of haloperidol and reduced haloperidol has been found to exacerbate the dystonic response produced by U50,488H (trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) -cyclohexyl]-benzeacetamide methane sulphonate) in guinea-pigs [8]. The present study sought to correlate the behavioural effect of haloperidol and reduced haloperidol with their effect on inhibition of sigma binding sites in guinea-pig brain using receptor binding and semi-quantitative autoradiography. In the first experiments, groups of guinea-pigs were injected with saline (control, n = 12), haloperidol (0.1 and 1 mg/kg i.p., n = 5) or reduced haloperidol (0.1 and 1 mg/kg i.p., n = 5) 1, 3 and 10 days before, followed by U50,488H (10 mg/kg s.c.) and the effect on the dystonic response rated using a behavioural rating scale [8]. In the second experiments, animals (n = 5) were injected with saline, haloperidol and reduced haloperidol as above and killed 1, 3 and 10 days later, their brains removed, dissected and tissue sections processed for sigma binding site autoradiography using [3H]3-(3-hydroxyphenyl)-N-(n-propyl)piperidine ([3H]-3-PPP). Triplicate tissue sections were wiped using GF/C filters and radioactivity counted. Injection of haloperidol and reduced haloperidol 1, 3 and 10 days earlier exacerbated the dystonic response by decreasing the latency to maximal dystonia and increasing the duration of the response at each dose tested compared with saline-treated animals. These effects of haloperidol and reduced haloperidol on latency and duration were time-related since the effect at 1 > 3 > 10 days. In addition, [3H]-3-PPP binding was inhibited by haloperidol and reduced haloperidol in a dose-and time-related manner. For example, % inhibition of [3H]-3-PPP binding for haloperidol (1 mg/kg) > haloperidol (0.1 mg/kg) and % inhibition of binding (mean +/- SEM) produced by haloperidol (0.1 mg/kg) at 1 (96.1 +/- 2.4) > 3 (74.8 +/- 4.8) > 10 days (36.2 +/- 1.6). Similar results were obtained for haloperidol (1 mg/kg) and reduced haloperidol (0.1 and 1 mg/kg). [3H]-3-PPP autoradiography confirmed these binding data. The results indicate that the exacerbation by sigma ligands of the dystonia produced by U50,488H was associated with the degree of inhibition of [3H]-3-PPP binding.
Asunto(s)
Analgésicos/farmacología , Haloperidol/farmacología , Pirrolidinas/farmacología , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Autorradiografía , Conducta Animal , Sitios de Unión , Femenino , Cobayas , Hipocampo/efectos de los fármacos , Masculino , Cloruro de Sodio/farmacología , Factores de TiempoRESUMEN
The present study examined whether the kappa-opioid agonists U50,488H (trans-(+/-)-3,4-dichloro-N-methyl-N[-2-(1-pyrrolidinyl)- cyclohexyl]-benzeacetamide methane sulphonate), bremazocine, spiradoline and ICI 197067 bind to sigma sites in guinea-pig tissues using in vitro, semi-quantitative receptor autoradiography and receptor binding, and compared the binding profile so obtained with those for several selective sigma ligands. Guinea-pigs were killed and their brians, livers and spleens were removed, tissue sections cut and processed for sigma binding site autoradiography using (+)-[3H]-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine ((+)-[3H]-3-PPP), or tissue was wiped and determined by liquid scintillation. Serial slide-mounted sections were incubated with 9-10 concentrations (1 nM-10 microM) of kappa opioids and their potency to inhibit (+)-[3H]-3-PPP binding compared with that of the sigma ligands haloperidol, DTG (1,3 di(o)-tolylguanidine), (+)-3-PPP, (+) and (-)pentazocine, SR 31742A and rimcazole (n = 3, duplicate determinations). Binding of (+)-[3H]-3-PPP to untreated, matched serial tissue sections was used as control. Kd values were estimated in brain, liver and spleen using quantitative, saturation binding analysis, IC50 values were determined from the binding data obtained by slide wiping experiments for each drug, and Ki values were calculated using the Cheng-Prussoff equation. All four kappa opioids inhibited (+)-[3H]-3-PPP binding to sigma 1-receptors with order of potency: brain: U50,488H = spiradoline > bremazocine > ICI 197067; liver: spiradoline > U50,488H > ICI 197067 > bremazocine; spleen: U50,488H > spiradoline > ICI 197067 > bremazocine. By comparison, the sigma ligands inhibited (+)-[3H]-3-PPP binding to matched, serial slide-mounted brain tissue sections (similar results in liver and spleen) with order of potency: SR 31742A > haloperidol > (+)pentazocine > (+)-3-PPP > DTG > (-)pentazocine > rimcazole. (+)-[3H]-3-PPP autoradiography confirmed these binding data. It is concluded that the kappa opioids tested moderately inhibit (+)-[3H]-3-PPP binding to sigma 1-receptors in guinea-pig brain, liver and spleen tissue with Ki values comparable to some selective sigma ligands and therefore are not opioid selective.
Asunto(s)
Encéfalo/efectos de los fármacos , Hígado/efectos de los fármacos , Receptores Opioides kappa/agonistas , Receptores sigma/efectos de los fármacos , Bazo/efectos de los fármacos , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Autorradiografía , Benzomorfanos/farmacología , Sitios de Unión , Encéfalo/metabolismo , Cobayas , Hígado/metabolismo , Piperidinas , Pirrolidinas/farmacología , Receptores sigma/metabolismo , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismo , Bazo/metabolismoRESUMEN
In guinea-pigs, acute treatment with mu and delta receptor opioid agonists induces sedation and immobility [1,5], and attenuates the behavioural activation produced by the dopamine D2 agonist quinpirole [5]. In contrast, kappa-selective opioid agonists induce dystonic-like movements [4,5,8]. This has led us to investigate the possibility of an interaction between acute opioid treatment and the dopamine D2 system. The effect of acute treatment with mu, delta and kappa opioid agonists on [3H]spiperone binding sites (dopamine D2) in guinea-pig brain was studied using receptor autoradiography. The mu preferring agonist morphine (15 mg/kg subcutaneously, SC) given for 2 h, and the delta receptor selective agonist DPDPE (Tyr-D-Pen-Gly-Phe-D-Pen) (20 nM, intracerebroventricularly, ICV) given for 0.5 h, both decreased the density of specific (butaclamol displaceable) [3H]spiperone binding in the caudate putamen by 23.8 +/- 1.7% and 24.2 +/- 2.7% respectively, and in nucleus accumbens by 26.1 +/- 2.7% and 21.9 +/- 4.6% respectively compared to saline treated animals. There were no significant changes in the level of [3H]spiperone binding to other brain regions examined including frontal cortex, hippocampus, substantia nigra, ventral tegmental area, amygdala, hypothalamic nuclei and cerebellum. In other experiments, incubation of coronal slices from various brain regions with [3H]spiperone, in the presence of a high concentration of morphine (20 microM) or DPDPE (10 microM) did not affect the level of binding, thus precluding effects due to residual tissue levels of drugs after in vivo treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Cuerpo Estriado/metabolismo , Receptores de Dopamina D2/metabolismo , Receptores Opioides delta/agonistas , Receptores Opioides kappa/agonistas , Receptores Opioides mu/agonistas , Espiperona/metabolismo , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Autorradiografía , Encéfalo/metabolismo , Encefalina D-Penicilamina (2,5) , Encefalinas/farmacología , Femenino , Cobayas , Masculino , Morfina/farmacología , Especificidad de Órganos , Pirrolidinas/farmacología , Receptores de Dopamina D2/efectos de los fármacos , TritioRESUMEN
Recent evidence suggests a role for sigma (sigma) binding sites in maintenance of cell growth and/or proliferation. The present study examines, for the first time, the effect of sigma binding site ligands on in vitro growth of tumour cells derived from human mammary adenocarcinoma (MCF-7, MDA) and colon carcinoma (LIM 1215, WIDr), and melanoma (Chinnery). Addition of the sigma ligands haloperidol, reduced haloperidol, 1,3-di(2-tolyl)guanidine (DTG), (+)- and (-)-N-allylnormetazocine (SKF 10,047), (+)- and (-)-pentazocine and rimcazole at 6.25, 12.5, 25, 50, 100 microM at the beginning of culture or 24 h later, inhibited cell proliferation in a dose-dependent manner. Light microscopy revealed cell detachment, rounding and cell death. The potency of sigma ligands on melanoma cells was rimcazole > reduced haloperidol > haloperidol = (+)-pentazocine, whereas DTG and (+)- and (-)-SKF 10,047 and (-)-pentazocine had no effect even at 100 microM. In contrast, in MCF-7 cells, rimcazole > reduced haloperidol > haloperidol > (-)-pentazocine > DTG > (+)-pentazocine > (+)-SKF 10,047 > (-)-SKF 10,047. For colon cancer cells, reduced haloperidol > DTG > haloperidol = (-)-pentazocine = (+)-pentazocine = (+)-SKF 10,047. Of all the ligands tested, rimcazole and reduced haloperidol were the most potent inhibitors of cell proliferation. With the exception of one slow-growing colon cancer cell line (LIM 1215), the order of sensitivity of various cell lines to reduced haloperidol, SFK 10,047, DTG, haloperidol and (+)- and (-)-pentazocine was colon carcinoma > mammary adenocarcinoma > melanoma, whereas to rimcazole, the sensitivities of mammary adenocarcinoma and melanoma cells were comparable. The effect of sigma ligands in MCF-7 and melanoma cells was not due to blockade of dopamine D1 and D2 receptors, serotonin (5-HT2) receptors, N-methyl-D-aspartate (NMDA)/phencyclidine receptors, beta-adrenoceptors or opioid receptors, since 100 microM SCH 23390, raclopride, mianserin, (+)-MK-801, propranolol and 1 microM naloxone respectively, were ineffective. However, mianserin and raclopride were inhibitory to melanoma cells and one colon carcinoma cell line, respectively. Taken together, the results are consistent with the recent observation that sigma binding sites may play a role in cell growth and/or cell proliferation.
Asunto(s)
Neoplasias de la Mama/patología , Neoplasias del Colon/patología , Melanoma/patología , Receptores sigma/metabolismo , Adenocarcinoma/patología , División Celular/efectos de los fármacos , Humanos , Ligandos , Células Tumorales CultivadasRESUMEN
The present study was undertaken to determine whether acute physical dependence occurred in guinea-pigs in vivo and guinea-pig isolated ileum following a single dose of the kappa-opioid receptor agonist U50,488H. Administration of naloxone hydrochloride, 15 and 30 mg/kg s.c., to guinea-pigs treated 1 h before with U50,488H, 10 mg/kg s.c., induced increased locomotor activity accompanied by behavioural responses which differed from those previously found in this species with morphine withdrawal. Nor-binaltorphimine, 10 mg/kg s.c., given 1 h after administration of U50,488H, 10 mg/kg s.c., produced a small but significant increase in locomotor activity but no other withdrawal behaviours. The morphine withdrawal response was not significantly affected by U50,488H, 1 or 10 mg/kg s.c. On the guinea-pig isolated ileum, nor-binaltorphimine, 1 microM, produced a withdrawal contracture following 2 min contact of the ileum with U50,488H 1 microM. U50,488H, 1 microM, abolished the [Met5]enkephalin withdrawal response of the ileum. It is concluded that dependence occurs following activation of kappa-opioid receptors, which is largely non-morphine-like in the central nervous system, but which is morphine-like in the enteric nervous system.
Asunto(s)
Analgésicos Opioides/farmacología , Pirrolidinas/farmacología , Receptores Opioides kappa/efectos de los fármacos , Trastornos Relacionados con Sustancias/etiología , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Conducta Animal/efectos de los fármacos , Femenino , Cobayas , Íleon/efectos de los fármacos , Técnicas In Vitro , Masculino , Dependencia de Morfina/fisiopatología , Actividad Motora/efectos de los fármacos , Contracción Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Naloxona/farmacología , Naltrexona/análogos & derivados , Naltrexona/farmacología , Síndrome de Abstinencia a Sustancias/fisiopatologíaRESUMEN
The sigma receptor ligands (+) and (-)pentazocine and BD1008 (1-100 microM) were added to rat forebrain synaptosomes. Their effects on intrasynaptosomal free calcium ([Ca2+(+)]i) levels under basal conditions and after depolarisation with high potassium buffer (45 mM KCl), veratridine (25 microM) and 4-aminopyridine (4-AP, 1 mM) were determined. The sigma ligands elicited significant, concentration-dependent decreases in basal [Ca2+]i levels with an order of potency (-)pentazocine > (+)pentazocine = BD1008. The sigma ligands (at the maximum effective concentrations) also significantly inhibited the rise in [Ca2+]i levels produced by depolarisation with KCl, veratridine and 4-AP. The effect of (+) and (-)pentazocine (100 microM) to inhibit the depolarisation-dependent increase in [Ca2+]i levels was greater when veratridine and 4-aminopyridine were used to depolarise the synaptosomes than with KCl, whereas the effect of BD1008 (100 microM) was approximately equipotent using all three depolarising agents. However, BD1008 was more potent to inhibit the KCl-induced rise in [Ca2+]i compared to (+) and (-)pentazocine. The data demonstrate for the first time that sigma ligands decrease [Ca2+]i levels in rat forebrain synaptosomes and this suggests a possible mechanism for the changes to neuronal protein phosphorylation and neurotransmitter release previously observed with sigma ligands.
Asunto(s)
Calcio/metabolismo , Prosencéfalo/metabolismo , Receptores sigma/metabolismo , Sinaptosomas/metabolismo , Animales , Bloqueadores de los Canales de Calcio/farmacología , Calibración , Etilaminas/farmacología , Colorantes Fluorescentes , Fura-2 , Técnicas In Vitro , Ligandos , Microscopía Fluorescente , Narcóticos/farmacología , Pentazocina/farmacología , Prosencéfalo/efectos de los fármacos , Pirrolidinas/farmacología , Ratas , Receptores sigma/agonistas , Sinaptosomas/efectos de los fármacosRESUMEN
The effects of sigma (sigma) ligands on protein phosphorylation were examined in crude, rat forebrain synaptosomes. Synaptosomes were prelabelled with 32P(i) and incubated with the sigma ligands 1,3-di-o-tolylguanidine (DTG), (+)pentazocine and (-)pentazocine (3, 10, 30, 100, 300 microM), or haloperidol, reduced haloperidol, and (+)SKF 10,047 (100 microM). Aliquots were then incubated for 10 s in control (5 mM K+) or depolarising buffer (41 mM K+). All the sigma ligands increased basal phosphorylation of synapsin Ib and other proteins including dynamin, and inhibited the depolarisation-dependent increase in phosphorylation of synapsin Ib in synaptosomes. The effects of these ligands are not directly on protein kinases or protein phosphatases. This indicates that the sigma ligands are mediating their effects via interaction with sigma binding sites, and suggest, for the first time, that protein phosphorylation may be one mechanism through which sigma ligands produce their biological effects.
Asunto(s)
GTP Fosfohidrolasas/metabolismo , Microtúbulos/metabolismo , Prosencéfalo/metabolismo , Receptores sigma/metabolismo , Sinapsinas/metabolismo , Sinaptosomas/metabolismo , Animales , Anticonvulsivantes/farmacología , Dinaminas , Guanidinas/farmacología , Haloperidol/farmacología , Técnicas In Vitro , Pentazocina/farmacología , Fenazocina/análogos & derivados , Fenazocina/farmacología , Fosforilación , RatasRESUMEN
Haloperidol, a 'typical' neuroleptic, with affinity for both dopamine (DA) receptors and sigma binding sites, exacerbates morphine withdrawal in guinea-pigs. In this study, the effects of sigma ligands (+)- and (-)-SKF 10,047 (1 and 10 mg/kg s.c.), pentazocine (20 mg/kg s.c.) and DTG (1 and 10 mg/kg s.c.), non-competitive NMDA antagonists ketamine hydrochloride (20 mg/kg s.c.) and MK-801 (0.025, 0.1 and 1 mg/kg s.c.), 'atypical' neuroleptic drugs with (remoxipride 25 mg/kg s.c.) and without (raclopride 10 mg/kg s.c.; clozapine 25 mg/kg s.c.) affinity for sigma sites, and atropine sulphate (20 mg/kg s.c.), were investigated on the opiate withdrawal response induced by naloxone hydrochloride (15 mg/kg s.c.) in guinea-pigs treated 2 h before with a single dose of morphine sulphate (15 mg/kg s.c.). (+)- and (-)-SKF 10,047, pentazocine, ketamine and MK-801, given 0.5 h before naloxone, significantly attenuated the increased locomotor activity and other behaviours associated with morphine withdrawal in guinea-pigs. The selective sigma ligand DTG, and remoxipride had no effect on the withdrawal response but raclopride, clozapine, and atropine exacerbated the response. It is concluded that exacerbation of the morphine withdrawal response by neuroleptics is not related to sigma activity but to other mechanisms. Furthermore NMDA but not sigma mechanisms might play a role in the morphine withdrawal response.
Asunto(s)
Antipsicóticos/farmacología , Morfina/efectos adversos , Receptores sigma/metabolismo , Síndrome de Abstinencia a Sustancias/metabolismo , Análisis de Varianza , Animales , Atropina/farmacología , Conducta Animal/efectos de los fármacos , Clozapina/farmacología , Maleato de Dizocilpina/farmacología , Femenino , Cobayas , Ketamina/farmacología , Cinética , Masculino , Actividad Motora/efectos de los fármacos , Naloxona/farmacología , Fenazocina/análogos & derivados , Fenazocina/farmacología , Racloprida , Ensayo de Unión Radioligante , Receptores sigma/efectos de los fármacos , Remoxiprida/farmacología , Salicilamidas/farmacologíaRESUMEN
Terminal osseous dysplasia and pigmentary defects is a rare X-linked dominant disorder with prenatal male lethality. Affected females display multiple systemic abnormalities such as limb deformities and pigmented lesions of the face and scalp. Phenotypic expression of the syndrome varies among the affected individuals. In this case report, we describe the syndromic dental and oral abnormalities in a female child aged 3 and 1/2 years. A widened bigonial width of the mandible and a brachyfacial pattern are observed. Intraoral findings include multiple frenulae, shallow mucobuccal fold, hypodontia, conical incisors, and other developmental structural defects.
Asunto(s)
Enfermedades del Desarrollo Óseo/patología , Anomalías de la Boca/patología , Trastornos de la Pigmentación/patología , Anomalías Dentarias/patología , Anomalías Múltiples , Anodoncia/patología , Enfermedades del Desarrollo Óseo/genética , Preescolar , Hipoplasia del Esmalte Dental/patología , Femenino , Humanos , Incisivo/anomalías , Frenillo Lingual/anomalías , Mandíbula/anomalías , Fenotipo , Trastornos de la Pigmentación/genética , SíndromeRESUMEN
PURPOSE: The objective of this pilot study was to determine the prevalence of coronal dental caries among children with different genetic sensitivity levels of taste, as determined by 6-n-propylthiouracil (PROP). METHODS: Coronal caries and restorations in permanent and primary dentition were evaluated in 150 healthy school-aged children aged 6 to 12 years. A filter paper containing 6-n-propylthiouracil was used to determine each subject's genetic ability to taste bitter and sweet substances. Supertasters perceived stronger tastes from a variety of bitter and sweet substances than both medium tasters and nontasters. The data were analyzed by ANOVA with Duncan's multiple range test, Mantel-Haenszel chi-square, multiple linear regression analyses, and Pearson's coefficient of correlation. RESULTS: The nontasters had more mean decayed, missing, and filled surfaces (dfs/DMFS) than tasters. The values of mean decayed and filled surfaces of primary dentition and mean decayed, missing, and filled surfaces of permanent dentition (dfs/DMFS) and mean decayed surfaces of primary dentition and permanent dentition (ds/DS) were significantly higher in nontasters than in medium tasters, and in medium tasters compared with supertasters. After adjusting for missing teeth, the data were expressed as a percentage of the available surfaces, and the significant differences in dfs/DMFS and ds/DS persisted (r = -0.49, P < .001 and r = -0.51, P < .0001 respectively). CONCLUSIONS: After all associated factors were controlled, taste was the only independent variable significantly related to overall caries experience. The results of this study suggested an increased prevalence of overall caries experience in nontaster children.
Asunto(s)
Antimetabolitos/química , Índice CPO , Propiltiouracilo/química , Umbral Gustativo/fisiología , Gusto/genética , Análisis de Varianza , Distribución de Chi-Cuadrado , Niño , Estudios Transversales , Caries Dental/clasificación , Restauración Dental Permanente , Femenino , Humanos , Modelos Lineales , Masculino , Proyectos Piloto , Estadística como Asunto , Umbral Gustativo/clasificación , Diente/patología , Pérdida de Diente/clasificación , Diente Primario/patologíaRESUMEN
The effect of naloxone-precipitated withdrawal after acute morphine was studied on the concentrations of noradrenaline (NA), 4-hydroxy-3-methoxyphenylethyleneglycol (MHPG), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), and on the metabolite/parent amine ratios MHPG/NA, DOPAC/DA and HVA/DA, in eight regions of the guinea-pig brain. Guinea-pigs were treated with a single dose of morphine sulphate (15 mg/kg s.c.) or saline (control) and 2h later with naloxone hydrochloride (15 mg/kg s.c.) to precipitate withdrawal. The animals were decapitated at 0.5 h or 1 h after naloxone injections and their brains analysed for monoamine concentrations by HPLC-ECD. At 0.5 h after naloxone-precipitated withdrawal NA and MHPG levels, and the MHPG/NA ratio, were increased in the hypothalamus, and the NA levels were increased in the hypothalamus, medulla/pons and cortex 1 h after naloxone. Naloxone-precipitated withdrawal also produced increased DA metabolism in the cortex, midbrain and medulla 0.5 h later, and in the cortex, hypothalamus and striatum 1 h later. Hence naloxone-precipitated withdrawal from acute morphine treatment produced a complex pattern of increased synthesis and metabolism of NA and DA which varied over time and with the brain region examined.