RESUMEN
The 18â kDa translocator protein (TSPO) is a ubiquitous conserved outer mitochondrial membrane protein implicated in numerous cell and tissue functions, including steroid hormone biosynthesis, respiration, cell proliferation, and apoptosis. TSPO binds with high affinity to cholesterol and numerous compounds, is expressed at high levels in steroid-synthesizing tissues, and mediates cholesterol import into mitochondria, which is the rate-limiting step in steroid formation. In humans, the rs6971 polymorphism on the TSPO gene leads to an amino acid substitution in the fifth transmembrane loop of the protein, which is where the cholesterol-binding domain of TSPO is located, and this polymorphism has been associated with anxiety-related disorders. However, recent knockout mouse models have provided inconsistent conclusions of whether TSPO is directly involved in steroid synthesis. In this report, we show that TSPO deletion mutations in rat and its corresponding rs6971 polymorphism in humans alter adrenocorticotropic hormone-induced plasma corticosteroid concentrations. Rat tissues examined show increased cholesteryl ester accumulation, and neurosteroid formation was undetectable in homozygous rats. These results also support a role for TSPO ligands in diseases with steroid-dependent stress and anxiety elements.
Asunto(s)
Hormona Adrenocorticotrópica/farmacología , Proteínas Portadoras/genética , Hidrocortisona/sangre , Polimorfismo de Nucleótido Simple , Receptores de GABA-A/genética , Receptores de GABA/genética , Adolescente , Adulto , Animales , Secuencia de Bases , Proteínas Portadoras/metabolismo , Ésteres del Colesterol/biosíntesis , Ésteres del Colesterol/sangre , Gonadotropina Coriónica/farmacología , Clonación Molecular , Corticosterona/biosíntesis , Corticosterona/sangre , Embrión de Mamíferos , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Humanos , Hidrocortisona/biosíntesis , Masculino , Plásmidos/química , Plásmidos/metabolismo , Pregnanolona/biosíntesis , Pregnanolona/sangre , Ratas , Ratas Transgénicas , Receptores de GABA/metabolismo , Receptores de GABA-A/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Testosterona/biosíntesis , Testosterona/sangre , Dedos de Zinc , Cigoto/efectos de los fármacos , Cigoto/crecimiento & desarrollo , Cigoto/metabolismoRESUMEN
The influence of stereochemistry and alkyl side chain length on the bioactivity of the Rho kinase inhibitor Y-27632 [(+)-1, R=Me] was examined by the synthesis of (+)- and (-)-1, and two alkyl chain analogs (+)- and (-)-2 (R=n-propyl) and (-)-3 (R=n-octyl) as well as their evaluation in enzymatic and neurite outgrowth assays.
Asunto(s)
Amidas/síntesis química , Inhibidores Enzimáticos/síntesis química , Neuritas/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Piridinas/síntesis química , Amidas/farmacología , Línea Celular , Inhibidores Enzimáticos/farmacología , Humanos , Péptidos y Proteínas de Señalización Intracelular , Neuritas/fisiología , Piridinas/farmacología , Estereoisomerismo , Relación Estructura-Actividad , Quinasas Asociadas a rhoRESUMEN
The Rho signaling pathway regulates the cytoskeleton and motility and plays an important role in neuronal growth inhibition. Here we demonstrate that inactivation of Rho or its downstream target Rho-associated kinase (ROK) stimulated neurite growth in primary cells of cortical neurons plated on myelin or chondroitin sulfate proteoglycan substrates. Furthermore, treatment either with C3 transferase (C3) to inactivate Rho or with Y27632 to inhibit ROK was sufficient to stimulate axon regeneration and recovery of hindlimb function after spinal cord injury (SCI) in adult mice. Injured mice were treated with a single injection of Rho or Rho-associated kinase inhibitors delivered in a protein adhesive at the lesion site. Treated animals showed long-distance regeneration of anterogradely labeled corticospinal axons and increased levels of GAP-43 mRNA in the motor cortex. Behaviorally, inactivation of Rho pathway induced rapid recovery of locomotion and progressive recuperation of forelimb-hindlimb coordination. These findings provide evidence that the Rho signaling pathway is a potential target for therapeutic interventions after spinal cord injury.