Nuclear Stiffness Decreases with Disruption of the Extracellular Matrix in Living Tissues.
Nuclear Stiffness Decreases with Disruption of the Extracellular Matrix in Living Tissues.
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DOI：
10.1002/smll.202006699
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发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Neu CP
中科院分区：
文献类型：
--
作者：
McCreery KP;Xu X;Scott AK;Fajrial AK;Calve S;Ding X;Neu CP
细胞核和细胞外基质之间的相互作用导致宏观组织表型变化。细胞外环境与核膜相连，并提供维持细胞稳态的线索，部分由机械转导机制调节。然而，很少有人知道细胞外基质环境如何影响基因表达和细胞表型在天然组织环境。研究核力学是深入了解基因调控和细胞病理学变化的一种方法，因为细胞核的力学性质会随着不同的基因调控机制而变化。以前，实验挑战限制了在细胞处于天然组织环境中时检测和直接测量核力学的能力。在这里，我们假设酶促破坏组织基质导致组织变软，影响嵌入细胞和核结构的刚度。我们的目标是直接测量核力学，而不干扰天然组织结构，以更好地了解核与细胞和组织微环境的相互作用。为了实现这一点，我们扩大了原子力显微镜针尖探针技术，探测培养细胞中的核硬度，以测量天然组织内的核膜和细胞膜硬度。我们通过成像针穿透和随后的稳定表达膜修复蛋白CHMP 4 B-GFP的HeLa细胞的质膜和核膜的修复来验证该技术。在离体天然组织环境中，我们发现，虽然用胶原酶3（MMP-13）和聚集蛋白聚糖酶-1（ADAMTS-4）酶促降解活软骨组织降低了组织基质硬度，但细胞和核膜硬度也降低。最后，我们展示了使用AFM针尖技术的细胞和细胞核弹性成像的能力。这些结果证明了传播到活细胞的质膜和内部核被膜结构的天然组织环境的破坏。细胞核是一种机械感应细胞器，其承受从细胞外基质传播的机械负荷，影响基因表达。在组织被酶促破坏后，应用原子力显微镜技术来测量细胞膜和核膜的硬度，同时细胞是有活力的并且仍然嵌入其天然组织环境中。
Reciprocal interactions between the cell nucleus and the extracellular matrix lead to macroscale tissue phenotype changes. The extracellular environment is linked to the nuclear envelope and provides cues to maintain cellular homeostasis regulated in part by mechanotransduction mechanisms. However, little is known about how the extracellular matrix environment affects gene expression and cellular phenotype in the native tissue environment. Studying nuclear mechanics is one way to gain insight into changes in gene regulation and cellular pathology, because the mechanical properties of the nucleus will change with different gene regulation mechanisms. Previously, experimental challenges limited the ability to detect and directly measure nuclear mechanics while cells are within the native tissue environment. Here, we hypothesized that enzymatic disruption of the tissue matrix results in a softer tissue, affecting the stiffness of embedded cell and nuclear structures. We aimed to directly measure nuclear mechanics without perturbing the native tissue structure to better understand nuclear interplay with the cell and tissue microenvironments. To accomplish this, we expanded an atomic force microscopy needle-tip probe technique that probes nuclear stiffness in cultured cells to measure the nuclear envelope and cell membrane stiffness within native tissue. We validated this technique by imaging needle penetration and subsequent repair of the plasma and nuclear membranes of HeLa cells stably expressing the membrane repair protein CHMP4B-GFP. In the native tissue environment ex vivo, we found that while enzymatic degradation of viable cartilage tissues with collagenase 3 (MMP-13) and aggrecanase-1 (ADAMTS-4) decreased tissue matrix stiffness, cell and nuclear membrane stiffness is also decreased. Finally, we demonstrated the capability for cell and nucleus elastography using the AFM needle-tip technique. These results demonstrate disruption of the native tissue environment that propagates to the plasma membrane and interior nuclear envelope structures of viable cells. The nucleus is a mechanosensing organelle that bears mechanical loads propagated from the extracellular matrix, impacting gene expression. After tissues are enzymatically disrupted, an atomic force microscopy technique is applied to measure stiffness of the cell membrane and nuclear envelope while cells are viable and still embedded in their native tissue environment.
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影响因子：
41.2
作者：
Chaudhuri O;Gu L;Klumpers D;Darnell M;Bencherif SA;Weaver JC;Huebsch N;Lee HP;Lippens E;Duda GN;Mooney DJ
通讯作者：
Mooney DJ
影响因子：
2.4
作者：
Korhonen, RK;Laasanen, MS;Jurvelin, JS
通讯作者：
Jurvelin, JS
影响因子：
1.6
作者：
HUTTER, JL;BECHHOEFER, J
通讯作者：
BECHHOEFER, J
影响因子：
4.1
作者：
Ashworth, JL;Murphy, G;Kielty, CM
通讯作者：
Kielty, CM
影响因子：
56.9
作者：
Jimenez, Ana Joaquina;Maiuri, Paolo;Perez, Franck
通讯作者：
Perez, Franck