**2.2. Methods**

#### *2.2.1. Electron microscopy*

For the electron microscopy the fixation of the specimens was performed in Sotelo's fixing solution, according to method, which was described in previous article [54]. Then they were post-fixed in 1% osmium tetroxide, dehydrated in graded alcohol solutions and propylene oxide [54]. Thin sections were cut in a Reichert ultratome, contrasted with uranyl acetate and lead citrate and studied in a Zeiss 9aS electron microscope [54].


The hypothalamus was excised and studied from 1974 to 2011.

AD: Alzheimer's disease, F: female, M: male. Fixation for silver impregnation techniques.

**Table 1.** List of the AD brains.

#### *2.2.2. Light microscope*

**2. Material and methods**

died accidentally, were used as normal controls [54].

lead citrate and studied in a Zeiss 9aS electron microscope [54].

The hypothalamus was excised and studied from 1974 to 2011.

**Table 1.** List of the AD brains.

AD: Alzheimer's disease, F: female, M: male. Fixation for silver impregnation techniques.

The morphological study of the hypothalamus concerns 14 autopsy cases of patients suffered from AD [54], at early stages according NINCDS-ADRDA criteria [55] and Braak and Braak staging [56] (**Table 1**). Twelve additional intact brains of apparently healthy individuals, who

Samples from the hypothalamus were excised and processed for electron microscopy and silver impregnation techniques including rapid Golgi's method, Rio Hortega's and Bodian's

For the electron microscopy the fixation of the specimens was performed in Sotelo's fixing solution, according to method, which was described in previous article [54]. Then they were post-fixed in 1% osmium tetroxide, dehydrated in graded alcohol solutions and propylene oxide [54]. Thin sections were cut in a Reichert ultratome, contrasted with uranyl acetate and

**Gender Age at death Duration of the disease Length of brain fixation in months Braak and Braak stage**

M 55 y 3 y 1 II/III F 62 y 28 mo 1 II/III M 63 y 37 mo 1 II F 66 y 40 mo 1 II/III M 72 y 3 y 1 III M 74 y 38 mo 1 II/III F 75 y 42 mo 1 II/III F 76 y 46 mo 1 III M 78 y 42 mo 1 II/III F 80 y 2 y 1 II/III M 78 y 42 mo 1 II/III F 76 y 36 mo 1 III M 54 y 2 y 1 III M 65 y 37 mo 1 II/III

**2.1. Material**

170 Cognitive Disorders

techniques [57, 58].

*2.2.1. Electron microscopy*

**2.2. Methods**
