Sox10 Is a Specific Biomarker for Neural Crest Stem Cells in Immunohistochemical Staining in Wistar Rats

Abstract

Objective: Neural crest stem cells (NCSCs) are prototypically migratory cells immigrating from the dorsal neural tube to specific embryonic sites where they generate a variety of cell types. A lot of biomarkers for NCSCs have been identified. However, which biomarkers are the most specific is still unclear.

Methods: The rat embryos harvested in embryonic day 9 (E9), E9.5, E10, E10.5, E11, E12, E13, and E14 were paraffin-embedded and sectioned in transverse. NCSCs were spatiotemporally demonstrated by immunohistochemical staining with RET, p75NTR, Pax7, and Sox10. NCSCs were isolated, cultured, and stained with RET, p75NTR, Pax7, and Sox10.

Results: In the paraffin sections of rat embryos, the immunohistochemical staining of RET, p75NTR, and Sox10 can all be used in demonstrating NCSCs. Sox10 was positive mainly in NCSCs while RET and p75NTR were positive not only in NCSCs but also in other tissue cells. In primary culture cells, Sox10 was mainly in the nucleus of NCSCs, RET was mainly in the membrane, and p75NTR was positive in cytoplasm and membrane.

Conclusions: Sox10 is the specific marker for immunohistochemical staining of NCSCs in paraffin sections. In cultured cells, Sox10, p75NTR, and RET presented a similar staining effect.

Figure 1

Comparison of immunohistochemical staining results for Sox10, Pax7, p75NTR, and RET in normal rat embryos of different developmental stages (E10 and E12). Scale bar, 100 μm.

Figure 2

The migratory pathway and pattern of NCSCs were observed with Sox10 as a marker. (a) E9, the neural tube has not been completely closed, and the neural crest has not been formed. (b) E9.5, the neural crest began to form, and NCSCs were delaminated. (c) E10, NCSCs migrated symmetrically to both sides along the gap between neural tube and epidermis. (d) E10.5, NCSCs migrated to both sides of salivary glands and chordates. (e) E11, NCSCs migrated to both sides of the dorsal aorta, and some cells entered the bilateral pericardial cavity. (f) E12, NCSCs were transplanted in the pericardial cavity, and some cells began to pass through the original diaphragm and enter the anterior intestine, middle intestine, pancreas, and spleen. (g) E13, NCSCs began to colonize the intestinal wall. (h) E14, ENS is formed. Scale bar, 100 μm.

Figure 3

Immunological staining for Sox10, Pax7, p75NTR, and RET in primary cell culture of ENCCs isolated from E12 normal rat embryos. Scale bar, 50 μm.