Page 16 - C. Biotechnology and molecular imaging

[C. Biotechnology and molecular imaging-10]

[Part 3] Investigation of Proto-oncogene PRDM Family

Expression Pattern during Mouse Embryonic Development by

using mPACT-A

Jiwon Woo¹˙²˙, Haewon Kang¹˙², Yong-Eun Cho¹˙²˙*

¹Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul 03722, Republic of Korea, ²The Spine

and Spinal Cord Institute, Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University College of
Medicine, Seoul 06273, Republic of Korea

Recent developments in tissue clearing methods such as CLARITY have allowed for the three-dimensional analysis

of biological structures in whole, intact tissue, providing greater understanding of spatial relationships and biological
circuits. Nonetheless, studies have reported issues with maintaining structural integrity and preventing tissue

disintegration, preventing the wide application of these techniques to fragile tissues such as developing embryos.
Here, we present optimized passive clearing techniques, mPACT-A, that improve tissue rigidity without the expense

of optical transparency. We also present a further modified mPACT-A protocol that is specifically optimized for
handling mouse embryos, which are small and fragile, such that they easily dismantle when processed via

established tissue clearing methods. We demonstrate proof-of-concept by investigating the expression of two
relatively understudied PRDM proteins, PRDM7 and PRDM12, in intact cleared mouse embryos at various stages of

development. We observed strong PRDM7, PRDM8, PRDM12 and PRDM13 expression in the developing mouse
nervous system, suggestive of potential roles in neural development that will be tested in future functional studies.