A key role for membrane transporter NKCC1 in mediating chondrocyte volume increase in the mammalian growth plate.
A key role for membrane transporter NKCC1 in mediating chondrocyte volume increase in the mammalian growth plate.
复制标题
DOI：
10.1002/jbmr.47
复制
发表时间：
2010-07
影响因子：
6.2
通讯作者：
Hall, Andrew C.
中科院分区：
文献类型：
--
作者：
Bush, Peter G.;Pritchard, Meredith;Loqman, Mohamad Y.;Damron, Timothy A.;Hall, Andrew C.
生长板软骨细胞体积增加，从而骨延长的机制知之甚少。许多细胞类型激活Na-K-Cl协同转运蛋白（NKCC），从而导致体积增加。我们推测NKCC可能是肥大软骨细胞体积扩张的原因。在存在/不存在特异性NKCC抑制剂布美他尼的情况下孵育来自16只大鼠幼崽（P7）的跖骨/掌骨，并在24小时后进行全骨长度测量和生长板组织学分析。使用共聚焦激光扫描显微镜在7只大鼠胫骨生长板（P7）上观察荧光NKCC免疫组织化学。对从三个年龄段（P49/53/58）的各五只大鼠的胫骨生长板的增殖和肥大区细胞分离的mRNA进行微阵列分析。暴露于布美他尼导致骨生长以剂量依赖性方式减少约35%（配对学生t检验，p <0.05）;组织学分析显示肥大区高度的减少是原因。荧光免疫组织化学的定量显示NKCC从增殖细胞的细胞内空间到肥大区细胞的胞质膜的显著（配对Student’s t检验，p <0.05）变化。此外，微阵列分析表明，在增殖和肥大细胞之间的NKCC 1 mRNA的增加。在NKCC抑制剂布美他尼存在下，肥大区细胞中NKCC 1 mRNA的增加、其细胞定位以及骨生长的减少暗示NKCC参与生长板肥大软骨细胞体积的增加。需要进一步研究以确定哺乳动物生长板中NKCC的调控以及长期暴露于髓袢利尿剂对骨生长的可能有害影响。© 2010美国骨与矿物质研究学会。
The mechanisms that underlie growth plate chondrocyte volume increase and hence bone lengthening are poorly understood. Many cell types activate the Na-K-Cl cotransporter (NKCC) to bring about volume increase. We hypothesised that NKCC may be responsible for the volume expansion of hypertrophic chondrocytes. Metatarsals/metacarpals from 16 rat pups (P7) were incubated in the presence/absence of the specific NKCC inhibitor bumetanide and measurement of whole-bone lengths and histologic analysis of the growth plate were done after 24 hours. Fluorescent NKCC immunohistochemistry was visualised using a confocal laser scanning microscopy on seven rat tibial growth plates (P7). Microarray analysis was performed on mRNA isolated from proliferative and hypertrophic zone cells of tibial growth plates from five rats of each of three ages (P49/53/58). Exposure to bumetanide resulted in approximately 35% reduction (paired Student's t test, p < .05) of bone growth in a dose-dependent manner; histologic analysis showed that a reduction in hypertrophic zone height was responsible. Quantification of fluorescence immunohistochemistry revealed a significant (paired Student's t test, p < .05) change in NKCC from the intracellular space of proliferative cells to the cytosolic membrane of hypertrophic zone cells. Further, microarray analysis illustrated an increase in NKCC1 mRNA between proliferative and hypertrophic cells. The increase in NKCC1 mRNA in hypertrophic zone cells, its cellular localization, and reduced bone growth in the presence of the NKCC inhibitor bumetanide implicate NKCC in growth plate hypertrophic chondrocyte volume increase. Further investigation is warranted to determine the regulatory control of NKCC in the mammalian growth plate and the possible detrimental effect on bone growth with chronic exposure to loop diuretics. © 2010 American Society for Bone and Mineral Research.
登录
查看更多内容
影响因子：
4.8
作者：
Satoh, H;Susaki, M;Hiraki, Y
通讯作者：
Hiraki, Y
影响因子：
3.2
作者：
Crouch, JJ;Sakaguchi, N;Schulte, BA
通讯作者：
Schulte, BA
影响因子：
3.6
作者：
KOO, WWK;GUAN, ZP;NEUMANN, V
通讯作者：
NEUMANN, V
影响因子：
4.2
作者：
Prandota, J
通讯作者：
Prandota, J
DOI：
10.1152/ajplung.00021.2004
发表时间：
2004-09-01
影响因子：
4.9
作者：
Iwamoto, LM;Fujiwara, N;Wada, RK
通讯作者：
Wada, RK