Immunolocalization of Sonic hedgehog in embryo-fetal mouse (Mus musculus) development

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Daniel Conei; Gustavo Saint-Pierre; Roberto Fierro; Mariano del Sol & Mariana Rojas Rauco

Summary

Sonic hedgehog (Shh) is an essential morphogen for the development of various structures, such as notochord, neural tube floor plate, limbs, among others. We sought to determine the immunolocalization of Shh in embryos and mouse fetuses. To do this, 10 pregnant mice (Mus musculus) BALB /c were euthanized, a group of 5 animals at 12.5 days post- coitus (dpc), and another group at 17.5 dpc. Embryos and fetuses obtained were fixed in 10 % formalin buffered in PBS and embedded in paraplast. Serial cross sections were made. Polyclonal antibody Shh (Santa Cruz Biotechnology, H-160, rabbit), dilution 1:100 was used. The immunolocalization of the positively labeled samples was identified and described. Shh expression in 12.5 dpc embryos was immunopositive in notochord, neural tube floor plate, radius precartilage and ulna, and practically all epithelia: bronchial, intestinal, bladder and urethra. In the fetal stage, at 17.5 dpc the immunopositivity disappears in the cartilage except for areas of ossification, decreases in the epidermis but appears in hair follicles. The intestinal mucosa has differentiated into segments, showing greater immunostaining at the level of the intestinal villi. Shh acts in different stages of the gestational period, being key in the differentiation of different structures. In embryonic stages, it is vital in the formation of the nervous system, organogenesis and formation of limbs, so its expression is found in these areas. However, in the fetal stage the expression changes to more specialized structures such as hair follicles and intestinal villi.

KEY WORDS: Sonic hedgehog; Morphogen; Organogenesis; Immunohistochemistry.

How to cite this article

CONEI, D.; SAINT-PIERRE, G.; FIERRO, R.; DEL SOL, M. & ROJAS, R. M. Immunolocalization of Sonic hedgehog in embryo-fetal mouse (Mus musculus) development. Int. J. Morphol., 36(2):693-698, 2018.