Hyper-SUMOylation of K+ Channels in Sudden Unexplained Death in Epilepsy: Isolation and Primary Culture of Dissociated Hippocampal Neurons from Newborn Mice for Subcellular Localization

Abstract

The physiological characteristics of rat and murine hippocampal neurons are widely studied, especially because of the involvement of the hippocampus in learning, memory, and neurological functions. Primary cultures of hippocampal neurons are commonly used to discover cellular and molecular mechanisms in neurobiology. By isolating and culturing individual hippocampal neurons, neuroscientists are able to investigate the activity of neurons at the individual cell and single synapse level, and to analyze properties related to cellular structure, cellular trafficking, and individual protein subcellular localization or protein-protein interaction using a variety of biochemical techniques. Conclusions addressed from such research are critical for testing theories related to memory, learning, and neurological functions. Here, we will describe how to isolate and culture primary hippocampal cells from newborn mice. The hippocampus may be isolated from newborn mice in as short as 2 min, and the cell cultures can be maintained for up to 2 weeks, and then ready for investigation of subcellular localization of K channel proteins and interaction with SUMO-specific protease 2 (SENP2). The protocol provides a fast and efficient technique for the culture of neuronal cells from mouce hippocampal tissue, and will ensure the immunocytochemistry detection of subcellular localization or protein-protein interactions in neurological research.

The function of SUMOylation and its crucial roles in the development of neurological diseases

“…After seizures, SENP2‐deficient mice develop atrioventricular block and cardiac arrest. Seizures and heart block are prevented by an opener of the Kv7 channel 15,19,159 . Therefore, we revealed the pathogenic role of SUMOylation in the nervous system and established an animal model of SUDEP (sudden unexplained death in epilepsy).…”

Schisandrin Restores the Amyloid β-Induced Impairments on Mitochondrial Function, Energy Metabolism, Biogenesis, and Dynamics in Rat Primary Hippocampal Neurons

“…Isolation of rat primary hippocampal neurons was conducted according to the previous reports [17]. Specially, the newborn Sprague-Dawley rats at the age of 1–3 days were obtained from Liaoning Changsheng Biotechnology Co., Ltd. (Liaoning, China) for the gathering of hippocampal tissues.…”

SUMOylation Wrestles With the Occurrence and Development of Breast Cancer

“…In recent years, additional studies have shown that SUMOylation and its pathways are associated with human diseases (25)(26)(27)(28). Various types of stress induce the upregulated SUMOylation, making SUMOylation a critical mechanism to protect cells from stress-induced apoptosis or cell death (29,30).…”

“…(sudden unexplained death in epilepsy) patients (25)(26)(27). SENP2 has been reported to play a crucial role in hepatocellular carcinoma (HCC) cell growth by modulating b-catenin stability (68).…”