Cloning and characterization of a novel gene encoding a putative seven-span transmembrane protein localized in endoplasmic reticulum

Objective: To clone and analyze the structure of a novel gene, named EST 1 (endoplasmic reticulum localized seven span transmembrane protein 1) and to analyze the expression pattern and intracellular location of EST 1. Methods: The cDNA library was screened to isolate novel cDNA fragment. The structure of novel gene was analysed by computer software. Expression of EST 1 was analyzed by dot blot and Northern blotting. Intracellular localization was observed after EST 1 enhanced green fluorescence protein (EGFP) fusion gene was transfected into mammalian cells. Results: The full length cDNA of mouse EST 1 was 1 802 bp, with a 1 293 bp open reading frame encoding 431 amino acids. It was predicated that protein encoded by EST 1 contained a signal peptide sequence at the N terminus, seven putative transmembrane domains, and an ER retaining signal at the C terminus. EST 1 EGFP fusion protein showed an ER like intracellular distribution in mammalian cells. Expression pattern analysis showed that EST 1 is expressed in all tissues examined. Conclusion: EST 1 is encoding a putative seven span transmembrane protein localized in endoplasmic reticulum. EST 1 was expressed in all tissues examined, suggesting an essential function of EST 1 in cells.

Local neuronal circuits that may shape the discharge patterns of inferior collicular neurons

The discharge patterns of neurons in auditory centers encode information about sounds.However,few studies have focused on the synaptic mechanisms underlying the shaping of discharge patterns using intracellular recording techniques.Here,we investigated the discharge patterns of inferior collicular（IC）neurons using intracellular recordings to further elucidate the mechanisms underlying the shaping of discharge patterns.Under in vivo intracellular recording conditions,recordings were obtained from 66 IC neurons in 18 healthy adult mice（Mus musculus,Km）under free field-stimulation.Fiftyeight of these neurons fired bursts of action potentials（APs）to auditory stimuli and the remaining eight just generated local responses such as excitatory（n=4）or inhibitory（n=4）postsynaptic potentials.Based on the APs and subthreshold responses,the discharge patterns were classified into seven types：phasic（24/58,41.4%）,phasic burst（8/58,13.8%）,pauser（4/58,6.9%）,phasic-pauser（1/58,1.7%）,chopper（2/58,3.4%）,primary-like tonic（14/58,24.1%）and sound-induced inhibitory（5/58,8.6%）.We concluded that（1）IC neurons exhibit at least seven distinct discharge patterns;（2）inhibition participates in shaping the discharge pattern of most IC neurons and plays a role in sculpting the pattern,except for the primary-like tonic pattern which was not shapedby inhibition;and（3）local neural circuits are the likely structural basis that shapes the discharge patterns of IC neurons and can be formed either in the IC or in lower-level auditory structures.