Apoptotic neurons in the hypothalamus of rats fed on high-fat diet.

(A–F) Transmission electron microscopy images of typical apoptotic neurons (A–E) and a normal neuron (F) in the arcuate nucleus of Wistar rats fed on high-fat (A, C–E) and control (B, F) diets, respectively. The arrows indicate typical apoptotic neurons. (G) Apoptotic neurons were counted in low magnification fields of transmission electron microscopy analysis from the arcuate nucleus of Wistar rats fed on high-fat (HF) and control (CT) diets, the results are presented as % of CT. (H) Representative synaptophysin immunofluorescence staining of samples from arcuate (Arc) and lateral hypothalamic (LH) nuclei, from Wistar rats fed on CT or HF diets. (I) Synaptophysin positive nerve terminals were field counted and the results are presented as % of CT. (J) Representative NeuN (rhodamine) and Bax (fluorescein) double immunoflorescence staining of samples from hypothalamus of Wistar rats fed on CT and HF diets; orange arrows depict neurons without Bax expression, yellow arrows depict double positive NeuN/Bax stained neurons. A–F are representative of n = 3; magnification, ×100 (scale bar, 20 µm), A–B; and ×20,000 (scale bar, 0.2 µm), C–F. G, field counting was performed in five distinct fields from n = 3; *p<0.05 vs. CT. H and J are representative of n = 5; magnification, ×100 (scale bar, 40 µm), H; and ×400 (scale bar, 10 µm), J; nuclei are stained in blue by DAPI. I, nerve terminals were counted in120 distinct fields (×200 magnification) from n = 5; *p<0.05 vs. CT.