Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy.
Altered synaptic connectivity in an in vitro human model of STXBP1 encephalopathy.
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DOI：
10.1093/brain/awac396
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发表时间：
2023-03-01
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影响因子：
--
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早期婴儿发育性脑病和癫痫性脑病是毁灭性的疾病，通常是遗传性的，但其病理机制往往仍不清楚。该研究领域的一个主要障碍是难以研究人类在子宫内相关时间段的皮质脑发育。为了解决这个问题，我们建立了一种体外测定法，通过使用人类亚板和邻近皮质区域的活器官型培养物来研究基因变异对人类大脑发育的影响，这些器官型培养物是从道德来源的受孕后 14-17 周的脑组织中制备的 (www.hdbr.org)。我们能够将培养物维持几个月，在此期间，皮质板、亚板和边缘区的总体解剖结构持续存在，而神经元继续在形态上发育并形成新的突触网络。因此，这种准备工作允许研究基因操作及其对完整发育中的人类皮质网络的下游影响。我们重点关注 STXBP1 单倍体不足，这是发育性脑病和癫痫性脑病最常见的遗传原因之一。这是通过 shRNA 干扰诱导的，导致突触功能受损和谷氨酸能突触密度降低。因此，我们为如何研究任何感兴趣的基因对人类皮质发育的影响提供了关键的原理验证。遗传性癫痫的病理学很可能始于子宫内。麦克劳德等人。使用由道德来源的组织样本制备的早期人类皮质的活器官切片培养物，建立一种体外测定法，研究特定基因变异对人类大脑发育的影响。
Early infantile developmental and epileptic encephalopathies are devastating conditions, generally of genetic origin, but the pathological mechanisms often remain obscure. A major obstacle in this field of research is the difficulty of studying cortical brain development in humans, at the relevant time period in utero. To address this, we established an in vitro assay to study the impact of gene variants on the developing human brain by using living organotypic cultures of the human subplate and neighbouring cortical regions, prepared from ethically sourced, 14–17 post-conception week brain tissue (www.hdbr.org). We were able to maintain cultures for several months, during which time the gross anatomical structures of the cortical plate, subplate and marginal zone persisted, while neurons continued to develop morphologically and form new synaptic networks. This preparation thus permits the study of genetic manipulations and their downstream effects on an intact developing human cortical network. We focused on STXBP1 haploinsufficiency, which is among the most common genetic causes of developmental and epileptic encephalopathy. This was induced using shRNA interference, leading to impaired synaptic function and a reduced density of glutamatergic synapses. We thereby provide a critical proof-of-principle for how to study the impact of any gene of interest on the development of the human cortex. The pathology of genetic epilepsies is likely to begin in utero. McLeod et al. establish an in vitro assay to study the impact of specific gene variants on the developing human brain, using living organotypic slice cultures of the early human cortex, prepared from ethically sourced tissue samples.
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影响因子：
16.6
作者：
He E;Wierda K;van Westen R;Broeke JH;Toonen RF;Cornelisse LN;Verhage M
通讯作者：
Verhage M
影响因子：
25
作者：
Weimer, RM;Richmond, JE;Jorgensen, EM
通讯作者：
Jorgensen, EM
影响因子：
14.5
作者：
Xian, Julie;Parthasarathy, Shridhar;Ruggiero, Sarah M.;Balagura, Ganna;Fitch, Eryn;Helbig, Katherine;Gan, Jing;Ganesan, Shiva;Kaufman, Michael C.;Ellis, Colin A.;Lewis-Smith, David;Galer, Peter;Cunningham, Kristin;O'Brien, Margaret;Cosico, Mahgenn;Baker, Kate;Darling, Alejandra;de Goes, Fernanda Veiga;El Achkar, Christelle M.;Doering, Jan Henje;Furia, Francesca;Garcia-Cazorla, Angeles;Gardella, Elena;Geertjens, Lisa;Klein, Courtney;Kolesnik-Taylor, Anna;Lammertse, Hanna;Lee, Jeehun;Mackie, Alexandra;Misra-Isrie, Mala;Olson, Heather;Sexton, Emma;Sheidley, Beth;Smith, Lacey;Sotero, Luiza;Stamberger, Hannah;Syrbe, Steffen;Thalwitzer, Kim Marie;van Berkel, Annemiek;van Haelst, Mieke;Yuskaitis, Christopher;Weckhuysen, Sarah;Prosser, Ben;Rigby, Charlene Son;Demarest, Scott;Pierce, Samuel;Zhang, Yuehua;Moller, Rikke S.;Bruining, Hilgo;Poduri, Annapurna;Zara, Federico;Verhage, Matthijs;Striano, Pasquale;Helbig, Ingo
通讯作者：
Helbig, Ingo
影响因子：
4.8
作者：
Parisotto, Daniel;Pfau, Maximilian;Soellner, Thomas H.
通讯作者：
Soellner, Thomas H.
DOI：
10.1038/nrn.2017.107
发表时间：
2017-10
期刊：
Nature reviews. Neuroscience
影响因子：
--
作者：
Di Lullo E;Kriegstein AR
通讯作者：
Kriegstein AR