NeuroanatomyResearch PaperA comparison of model-based (2D) and design-based (3D) stereological methods for estimating cell number in the substantia nigra pars compacta (SNpc) of the C57BL/6J mouse
Section snippets
Animals
Adult 3–4 month old male and female C57BL/6J mice (Jackson Laboratories, Bar Harbor, ME, USA) were used in this study. All studies were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publications No. 80–23) revised 1996 and were approved by the SJCRH Animal Care and Use Committee. The authors further attest that all efforts were made to minimize the number of animals used and their suffering.
Serial reconstruction of the SNpc
Eight 3–4 month old C57BL/6J
Serial reconstruction of the DA neurons in the SNpc of the C57BL/6J mouse
The total bilateral number of DA neurons (±SEM) in the SNpc from the four serially reconstructed brains was 8305±539 (n=4, range 7464–9763). On average, the rostral to caudal length of the paraffin embedded SNpc was 1150 μm. The distribution of the cells demonstrates that the majority of cells are present 400–700 μm into the SNpc (rostral to caudal) (Fig. 3). This is similar to the distribution of C57BL/6J SNpc DA neurons previously reported (Nelson et al 1996, Hamre et al 1999).
Model-based (2D) stereological estimates of DA neurons in the C57BL/6J SNpc
Model-based
Discussion
Establishment of a gold standard for the number of neurons within the SNpc of mice is necessary in order for investigators to both assess the accuracy of the estimated values of the given experiments and allow comparisons between experiments. This is especially important with regard to determining the efficacy of particular treatments that purport to cause or prevent neuronal loss. Here we used serial reconstruction of the entire adult SNpc of the C57BL/6J mouse to reveal that it contains
Conclusion
In conclusion, we believe that the empirically-determined number of DA neurons of the SNpc reported here, along with the multiple methods for estimating this value will be beneficial to many using this system to assess disease models and potential treatments. Surprisingly, we found that the accuracy and precision of the design-based stereological methods were equal to that of the method-based analysis. Thus, since both methods can be used in obtaining accurate neuronal estimates, one needs to
Acknowledgments
The authors acknowledge support from the National Parkinson's Foundation, NIH (NS39006 to R.J.S.) and P-30 CA 21765 and the American Lebanese Syrian Associated Charities (ALSAC). The authors thank Haeman Jang for assistance with photomicroscopy and Yun Jiao for assistance with histology. We thank the staff at MicroBrightField Inc., especially Dan Peruzzi, for comments on this paper.
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