Cathepsin B inhibition blocks neurite outgrowth in cultured neurons by regulating lysosomal trafficking and remodeling.
Cathepsin B inhibition blocks neurite outgrowth in cultured neurons by regulating lysosomal trafficking and remodeling.
复制标题
DOI：
10.1111/jnc.15032
复制
发表时间：
2020-11
影响因子：
4.7
通讯作者：
Ni J
中科院分区：
文献类型：
--
作者：
Jiang M;Meng J;Zeng F;Qing H;Hook G;Hook V;Wu Z;Ni J
已知溶酶体介导神经元中的神经突生长。然而，控制这种生长的主要溶酶体分子尚不清楚。我们在体外研究了原代小鼠神经元，发现它们随着时间的推移自然地发育出神经突，并且随着它们的发育，溶酶体半胱氨酸蛋白酶组织蛋白酶B（CTSB）mRNA水平显著增加。令人惊讶的是，我们发现用抑制CTSB的CA-074 Me处理这些神经元可以防止神经突起。由于该化合物还抑制另一种蛋白酶，我们使用CRISPR技术评估了其中CTSB基因缺失（CTSB KO）的N2 a神经元细胞系，并通过用视黄酸处理诱导其神经突生长。我们发现CTSB KO N2 a细胞未能产生神经突生长，但野生型（WT）产生。CA-074 Me是CA-074的细胞渗透性前药，CA-074是细胞不可渗透的，并且是特异性CTSB抑制剂。在分别用CA-074 Me和CA-074处理的WT N2 a细胞中，神经突生长受到抑制并且没有受到抑制。溶酶体相关膜糖蛋白2（LAMP 2）阳性溶酶体在WT中运输至浆细胞膜，但在CTSB KO N2 a细胞中未运输。有趣的是，在WT和CTSB KO N2 a细胞之间没有发现神经突生长调节蛋白、PI 3 K/AKT、ERK/MAPK、cJUN和CREB的明显差异。这些发现表明，细胞内CTSB控制神经突生长，它通过调节溶酶体运输和神经元重塑。这增加了关于CTSB在神经发育中的生理功能的有价值的信息。
Lysosomes are known to mediate neurite outgrowth in neurons. However, the principal lysosomal molecule controlling that outgrowth is unclear. We studied primary mouse neurons in vitro and found that they naturally develop neurite outgrowths over time and as they did so the lysosomal cysteine protease cathepsin B (CTSB) mRNA levels dramatically increased. Surprisingly, we found that treating those neurons with CA-074Me, which inhibits CTSB, prevented neurites. As that compound also inhibits another protease, we evaluated a N2a neuronal cell line in which the CTSB gene was deleted (CTSB KO) using CRISPR technology and induced their neurite outgrowth by treatment with retinoic acid. We found that CTSB KO N2a cells failed to produce neurite outgrowths but the wild-type (WT) did. CA-074Me is a cell permeable prodrug of CA-074, which is cell impermeable and a specific CTSB inhibitor. Neurite outgrowth was and was not suppressed in WT N2a cells treated with CA-074Me and CA-074, respectively. Lysosome-associated membrane glycoprotein2 (LAMP2)-positive lysosomes traffic to the plasma cell membrane in WT but not in CTSB KO N2a cells. Interestingly, no obvious differences between WT and CTSB KO N2a cells were found in neurite outgrowth regulatory proteins, PI3K/AKT, ERK/MAPK, cJUN and CREB. These findings show that intracellular CTSB controls neurite outgrowth and that it does so through regulation of lysosomal trafficking and remodeling in neurons. This adds valuable information regarding the physiological function of CTSB in neural development.
登录
查看更多内容
影响因子：
4.8
作者：
López-Carballo, G;Moreno, L;Barettino, D
通讯作者：
Barettino, D
DOI：
10.1016/s0169-328x(00)00191-1
发表时间：
2000-11-10
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Dragunow, M;Xu, RA;During, M
通讯作者：
During, M
DOI：
10.1007/978-1-4939-7595-2_12
发表时间：
2018-01-01
期刊：
PROTEASES AND CANCER: METHODS AND PROTOCOLS
影响因子：
--
作者：
Gnanamony, Manu;Gondi, Christopher S.
通讯作者：
Gondi, Christopher S.
影响因子：
--
作者：
Masia, Susana;Alvarez, Susana;Barettino, Domingo
通讯作者：
Barettino, Domingo
影响因子：
4.4
作者：
Ha, Soon-Duck;Martins, Andrew;Kim, Sung Ouk
通讯作者：
Kim, Sung Ouk