RESUMO
GEC1 protein shares high identity with GABARAP (GABA(A) Receptor-Associated Protein), interacts with tubulin and GABA(A) receptors and is potentially involved in intracellular transport processes. Recently, using quantitative real time PCR, we have reported the gec1 mRNA expression in different rat brain areas. In the present study, we investigated the cell types expressing gec1 in rat brain. Sense and anti-sense gec1 RNA probes, corresponding to the 3'-untranslated region, were generated. In northern blotting experiments, the anti-sense probe revealed only the 1.75 kb gec1 mRNAs. On the other hand, in immunohistochemistry experiments, GEC1 polyclonal antibodies did not discriminate between GEC1 and GABARAP proteins. Therefore, we used digoxigenin-labeled RNA probes for in situ hybridization (ISH) experiments to map the gec1 expression. Using the anti-sense probe, we detected the gec1 mRNAs specifically in neurons throughout the rostrocaudal extent of the brain as well as in the spinal cord. Although a majority of neurons expressed the gec1 mRNAs, different intensities of labeling were observed depending on the areas: the strongest labeling was observed in the isocortex, hippocampus, basal telencephalon, some thalamic and most of hypothalamic nuclei, cerebellum, and numerous brainstem nuclei. Furthermore, the gec1 mRNAs were intensely expressed in neurons involved in somatomotor and neuroendocrine functions and weakly expressed in sensory and reticular structures. These results corroborate the putative role of the GEC1 protein in the trafficking of receptor GABA(A).
Assuntos
Encéfalo/metabolismo , Proteínas de Transporte/genética , Proteínas Associadas aos Microtúbulos/genética , RNA Mensageiro/metabolismo , Receptores de GABA-A/metabolismo , Medula Espinal/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Elementos Antissenso (Genética) , Encéfalo/anatomia & histologia , Mapeamento Encefálico , Proteínas de Transporte/biossíntese , Hibridização In Situ , Masculino , Proteínas Associadas aos Microtúbulos/biossíntese , Vias Neurais/anatomia & histologia , Vias Neurais/metabolismo , Neurônios/metabolismo , Ratos , Ratos Sprague-Dawley , Medula Espinal/anatomia & histologiaRESUMO
GABARAP and GEC1/GABARAPL1 interact with tubulin and GABA(A) receptor and belong to a new protein family. This family includes GATE 16 and LC3, potentially involved in intracellular transport processes. In this study, we combined brain dissection and quantitative real-time reverse transcription polymerase chain reaction to study discriminatively gabarap, gec1/gabarapL1, gate16/gabarapL2, lc3 mRNA distribution in multiple rat brain areas.
