Interlichia Jillian P, Williams Nolann G, Rodgers Buel D
Department of Animal Sciences, Washington State University, Pullman, WA 99164, USA.
Growth Horm IGF Res. 2010 Apr;20(2):171-3. doi: 10.1016/j.ghir.2009.10.004. Epub 2009 Nov 27.
One of the most accurate indices of changes in somatic tissue growth rate in rodents is the width of tibial epiphyseal plates as unlike most mammals, rodent growth plates never ossify. Unfortunately, the original procedure to measure tibial epiphyseal plate width (TEPW) was developed for rats and yields poor results with mice. This paper demonstrates a simple method for silver staining growth plates that can be used to inexpensively and quickly measure the TEPW of mice. Poor visualization due to overstaining and the shattering of growth plates necessitated several revisions to the original protocol. These include exposing the growth plate prior to acetone dehydration, reducing the silver nitrate concentration from 2% to 1.5% and staining time from 2 min to 10 s and finally, the use of reflective light rather than transmissive light when imaging. The optimized protocol was then validated by generating an age-dependent TEPW growth curve that matched changes in tibia length. A total of 120 tibias were processed in a combined time of less than one day and for less than $30. By contrast, histological processing in the university's core facility would have cost $1440 and taken approximately three weeks. Thus, the revised protocol is vastly more cost effective, reliable and can be performed considerably quicker with minimal training.
啮齿动物体细胞组织生长速率变化最准确的指标之一是胫骨骨骺板的宽度，因为与大多数哺乳动物不同，啮齿动物的生长板从不骨化。不幸的是，最初测量胫骨骨骺板宽度（TEPW）的方法是针对大鼠开发的，对小鼠的测量结果不佳。本文展示了一种用于生长板银染的简单方法，可用于廉价且快速地测量小鼠的TEPW。由于过度染色导致的可视化不佳以及生长板破碎，需要对原始方案进行多次修订。这些修订包括在丙酮脱水前暴露生长板，将硝酸银浓度从2%降至1.5%，染色时间从2分钟降至10秒，最后，成像时使用反射光而非透射光。然后通过生成与胫骨长度变化相匹配的年龄依赖性TEPW生长曲线来验证优化后的方案。总共120根胫骨在不到一天的时间内处理完成，花费不到30美元。相比之下，在大学核心设施进行组织学处理将花费1440美元，大约需要三周时间。因此，修订后的方案成本效益更高、更可靠，并且只需最少的培训就能更快地完成。
