Journal: Human Molecular Genetics

Article Title: EAP1 regulation of GnRH promoter activity is important for human pubertal timing

doi: 10.1093/hmg/ddy451

Figure Lengend Snippet: Eap1 mRNA is expressed in the hypothalamus of peri-pubertal mice. Peri-pubertal mice hypothalamus sections were used for in situ hybridization and immunohistochemistry to localize Eap1 mRNA (staining in purple) and GnRH neurons (staining in brown), respectively. Eap1 is expressed in the VMH, Arc and Pa nuclei of (A) male and (B) female mice. GnRH neuron bodies are dispersed predominantly in the medial preoptic area (C) in an Eap1 positive milieu. GnRH neuron bodies are shown at higher magnification in (D) . GnRH neuron projections are also detected in the ME (A) and at the level of the 3V (C). Sense probe was used as a negative control with no detectable staining (E). Arrows indicate GnRH neuron bodies. ♀ indicates female; ♂, male; 3V, third ventricle; AHC, anterior hypothalamic area, central part; MPA, medial preoptic area; VMPO, ventromedial preoptic nucleus.

Article Snippet: When co-labeling was desired, after in situ hybridization, the sections were incubated with primary antibodies (anti-GnRH; Immunostar, Hudson, Wisconsin, USA) diluted at 1:1000 in PBS–Triton 0.1%, as used in ( ).

Techniques: In Situ Hybridization, Immunohistochemistry, Staining, Negative Control