Septal innervation of basal forebrain cholinergic neurons to the hippocampus is critical for normal learning and memory and is severely degenerated in Alzheimer’s disease. To understand the molecular events underlying physiological cholinergic synaptogenesis and remodeling, as well as pathological loss, we developed an optimized primary septal-hippocampal co-culture system. Hippocampal and septal tissue were harvested from embryonic Sprague–Dawley rat brain and cultured together at varying densities, cell ratios, and in the presence of different growth factors. We identified conditions that produced robust septal-hippocampal synapse formation. We used confocal microscopy with primary antibodies and fluorescent ligands to validate that this system was capable of generating developmentally mature cholinergic synapses. Such synapses were comprised of physiological synaptic partners and mimicked the molecular composition of in vivo counterparts. This co-culture system will facilitate the study of the formation, plasticity, and dysfunction of central mammalian cholinergic synapses.

基底前脑胆碱能神经元到海马的间隔神经支配对于正常学习和记忆至关重要，并且在阿尔茨海默病中严重退化。为了了解生理胆碱能突触发生和重塑以及病理损失的分子事件，我们开发了一种优化的原代间隔-海马共培养系统。从胚胎 Sprague-Dawley 大鼠脑中采集海马和间隔组织，并以不同的密度、细胞比例和不同的生长因子一起培养。我们确定了产生强大的间隔海马突触形成的条件。我们使用具有一抗和荧光配体的共聚焦显微镜来验证该系统能够产生发育成熟的胆碱能突触。这种突触由生理突触伙伴组成，并模仿体内对应物的分子组成。这种共培养系统将有助于研究中枢哺乳动物胆碱能突触的形成、可塑性和功能障碍。