Spinal cord nerve fiber projection is repressed by sonic hedgehog overexpression during chicken embryonic development

doi:10.16098/j.issn.0529-1356.2015.01.001

Abstract

Abstract:

Objective To study the effect of sonic hedgehog(Shh) overexpression on nerve fiber projection in the spinal cord during chicken embryonic development.
Methods At embryonic day(E)2.5-3.0, in vivo electroporation was used to co-transfect Shh expression plasmid and reporter gene plasmid into the chicken embryonic spinal cord. Samples were collected and sliced at E6. At least three samples were collected. The overexpression of Shh was detected using immunofluorescence histochemistry, and nerve fiber projecting affected by Shh overexpression was detected with Open Book technology. Results The results of immunofluorescence showed Shh was overexpressed in the chicken embryonic spinal cord successfully. Compared with the control group, after Shh overexpression, the nerve fiber did not across the floor plate and project stereotypically both in slice level and whole mount level, which suggested that Shh plays an important role for nerve fiber projecting in the chicken embryonic spinal cord. Conclusion The normal nerve fiber projection is repressed by Shh overexpression in the chicken embryonic spinal cord.

Key words: Sonic hedgehog, Nerve fiber projecting, In vivo electroporation, Open Book, Chicken embryo

LI Han YANG Ci-qing WANG Cong-rui HU A-zhen FU Su-lei LIN Jun-tang*. Spinal cord nerve fiber projection is repressed by sonic hedgehog overexpression during chicken embryonic development[J]. AAS, 2015, 46(1): 1-5.

References

［1］Bovolenta P, Sanchez-Arrones L.Shh goes multidirectional in axon guidance［J］.Cell Res, 2012, 22(4):611-613.
［2］Charron F, Stein E, Jeong J, et al. The morphogen sonic hedgehog is an axonal chemoattractant that collaborates with netrin-1 in midline axon guidance［J］.Cell,2003,113(1): 11-23.
［3］Bourikas D, Pekarik V, Baeriswyl T, et al. Sonic hedgehog guides commissural axons along the longitudinal axis of the spinal cord［J］.Nat Neurosci,2005,8(3):297-304.
［4］Parra LM, Zou Y. Sonic hedgehog induces response of commissural axons to Semaphorin repulsion during midline crossing［J］.Nat Neurosci,2010,13(1):29-35. 
［5］Yang CQ, Mao HL, Lin JT. Establishment of chicken embryo culture system and in vivo electroporation methods［J］. Acta Anatomica Sinica, 2012,43(4):564-568.（in Chinese）
杨慈清，毛会丽，林俊堂. 鸡胚培养及活体电转基因方法的建立［J］.解剖学报，2012，43（4）:564-568.
［6］Hu AZh, Yang CQ, Fu SL, et al. A new method to study nerve fiber projecting in the spinal cord during chicken embryo development［J］. Acta Anatomica Sinica, 2014,45(5): 598-602.（in Chinese）
胡阿珍，杨慈清，付苏雷, 等. 一种鸡胚发育过程脊髓神经纤维投射研究方法的建立［J］.解剖学报，2014,45（5）:598-602.
［7］Liu F, Placzek M, Xu H. Axon guidance effect of classical morphogens Shh and BMP7 in the hypothalamo-pituitary system［J］.Neurosci Lett,2013,553:104-109.
［8］Okada A, Charron F, Morin S, et al. Boc is a receptor for sonic hedgehog in the guidance of commissural axons［J］.Nature, 2006, 16;444(7117):369-373.
［9］Choudhry Z, Rikani AA, Choudhry AM, et al. Sonic hedgehog signalling pathway: a complex network［J］. Ann Neurosci, 2014, 21(1):28-31.
［10］Avilés EC, Wilson NH, Stoeckli ET. Sonic hedgehog and Wnt: antagonists in morphogenesis but collaborators in axon guidance［J］. Front Cell Neurosci, 2013, 7(86):1-17.
［11］Domanitskaya E, Wacker A, Mauti O, et al. Sonic hedgehog guides post-crossing commissural axons both directly and indirectly by regulating Wnt activity［J］. J Neurosci,2010,30(33):11167-11176.
［12］Sánchez-Camacho C, Bovolenta P. Emerging mechanisms in morphogen-mediated axon guidance［J］. Bioessays,2009, 31(10):1013-1025.
［13］Sakai N, Kaprielian Z. Guidance of longitudinally projecting axons in the developing central nervous system［J］.Front Mol Neurosci,2012,5(59):1-14.
［14］Sakai N, Insolera R, Sillitoe RV, et al. Axon sorting within the spinal cord marginal zone via Robo-mediated inhibition of N-cadherin controls spinocerebellar tract formation［J］.J Neurosci,2012,32(44):15377-15387.

[1]	LI Qiu-ling YANG Ci-qing ZHANG Bi-chao ZHU Shao-yi LIN Jun-tang. Construction of Wnt3a fusion protein vector and its effect on the proliferation and axon formation of neural precursor cells during the chick embryonic spinal cord development [J]. Acta Anatomica Sinica, 2017, 48(1): 7-13.
[2]	CAI Yu-jin LI Hui-chao ZHANG Hong-quan JING Ya* YANG Yan-ping CUI Hui-lin. Expression of sonic hedgehog and islet-1 in morphogenesis of respiratory system in mouse embryo [J]. AAS, 2016, 47(5): 678-682.
[3]	LI Han YANG Ci-qing QIAO Liang ZHANG Bi-chao LIN Jun-tang*. Influence of sonic hedgehog overexpression on Pax family in the spinal cord during chicken embryonic development [J]. AAS, 2015, 46(5): 667-672.
[4]	YANG Ci-qingLI Xiao-ying WANG Cong-rui LI Han ZHANG Bi-chao LIN Jun-tang*. A self-made electrode used for in vivo electroporation in chicken embryonic tectum [J]. AAS, 2015, 46(1): 133-137.
[5]	HU A-zhen1 YANG Ci-qing FU SU-lei JIA Yang-yang LI Han GUO Zhi-kun LIN Jun-tang *. A new method to study nerve fiber projecting in the spinal cord during chicken embryo development [J]. AAS, 2014, 45(5): 724-728.
[6]	LI Hui-chao JING Ya* SHI Liang YANG Yan-ping LI Hai-rong QIAO Cong-jin QIAO Ai-xiu CAI Yu-jin. Association of morphogenesis of pulmonary endoderm with development of prepharyngeal mesenchyme and outflow tract septation in mouse embryos [J]. AAS, 2013, 44(6 ): 804-811.
[7]	. Establishment of chicken embryo culture system and EM>in vivo/EM> electroporation methods [J]. , 2012, 43(4): 564-568.
[8]	. Differentiation of rat bone marrow mesenchymal stem cells into dopaminergic neurons after treatment with sonic hedgehog and fibroblast growth factor-8 [J]. , 2011, 42(1): 22-26.
[9]	. Temporal-spatial expression of SHH and β-catenin during hair follicle morphogenesis in different hoof periphery site of EM>Bos grunniens/EM> embryos [J]. , 2009, 40(5): 816-820.
[10]	. Effect of sonic hedgehog on the proliferation and differentitation of neural stem cells [J]. , 2009, 40(2): 211-214.
[11]	. EFFECT OF EphB/Ephrin-B1 SIGNAL ON THE CURRENTS OF α7-nAChRs ON CILIARY GANGLION NEURONS [J]. , 2008, 39(5): 656-660.
[12]	. DERIVATION OF ADIPOCYTES AND NEURONLIKE CELLS FROM CHICKEN EMBRYONIC PRIMORDIAL GERM CELLS IN VITRO [J]. , 2008, 39(4): 531-538.