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An immunohistochemistry protocol to identify satellite glial cells positive for GFAP in the dorsal root ganglion of rats in different ages
Author(s) -
Simões Ana Leda B,
Giorgetto Carolina,
Carmo Elisabete C,
Machado Nathalia B,
Sanada Luciana S,
Fazan Valeria Paula Sassoli
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.564.2
Subject(s) - glial fibrillary acidic protein , dorsal root ganglion , immunostaining , immunohistochemistry , biology , pathology , paraformaldehyde , ganglion , anatomy , dorsum , medicine
The nervous system is characterized by its high capacity of differentiation and cellular plasticity, especially during the neonatal period [1]. It is well known that the satellite glial cells (SGCs) in the dorsal root ganglion of adult animals are easily identified by their expression of glial fibrillary acidic protein (GFAP), and surround the neuron cell body forming a typical “ring shape” in histological images. However, we didn't find any description in the literature regarding the development and maturation of satellite glial cells in the dorsal root ganglion (DRG) The aim of this work is to validate an immunohistochemical method (IHQ), which identifies SGCs in DRG of rats during different stages of development. Thus, Wistar rats (n=4), male and female, were investigated 15, 30, 90 and 180 days after birth. The animals were anesthetized, perfused with 4% buffered paraformaldehyde and had the left L5 DRG removed and frozen. Criossections (8 μm) were incubated in rabbit polyclonal antibody anti‐glial fibrillary acidic protein (GFAP) (1:2.000, 24 hr) followed by biotinylated anti‐rabbit IgG (1:500; 24h) and finally Extravidin‐HRP (1:1.500; 24h) [2]. Our results showed that there was immunoreactivity for small round cells in the DRG of animals with 15 and 30 days‐old, whereas there was immunostaining in SGCs of animals with 90 and 180 days‐old, according to the patterns described in the literature. This indicates that his method is viable, reproducible and effective for the identification of SGCs in the DRG of adult animals. We also demonstrated that the hitological organization of the SGCs in the DRG as described in the literature appears only after 90 days of life. Future studies will be performed to investigate this development and maturation of the SGCs in the DRG. Support or Funding Information FAPESP, CNPq, CAPES and FAEPA