Suprachiasmatic nucleus of the human brain: An immunocytochemical and morphometric analysis

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Abstract
Background The present paper describes the immunocytochemical and morphometric characteristics of two major cell groups of the suprachiasmatic nucleus (SCN) in the human hypothalamus: the vasopressin (VP) and vasoactive intestinal polypeptide (VIP) neuronal subdivisions. The dimensions (volume and length) and the number of neurons expressing each peptide in the two subdivisions were obtained, as well as the mean diameter of the cell nuclei. All morphometric parameters were studied in relation to sex and age. Methods Brains of 42 human subjects (22 males and 20 females) ranging in age from 10 to 92 years were obtained at autopsy. The hypothalamic area containing the SCN was dissected from each brain, dehydrated, and embedded in paraffin. Serial sections of 6 μm were cut in a coronal plane and stained with thionin for general orientation. To determine the architectonic boundaries of the VP‐ and VIP‐expressing cell populations every 25th section was immunocytochemically stained by means of antibodies against arginine VP or VIP using the peroxidase‐antiperoxidase method. The VP‐ and VIP‐expressing cell numbers in the SCN of each subject were estimated by unilaterally counting the number of nuclear profiles with the aid of a Zeiss microscope under ×500 magnification, using a deconvolution procedure and a correction for section thickness. Results The main portion of the VP positive neurons is located in the dorsomedial part of the SCN and is rostrocaudally longer in females than in males (1.76 ± 0.12 mm and 1.40 ± 0.10 mm, respectively). The volume of the VP subdivision is 0.244 ± 0.017 mm3 and contains 6,890 ± 520 VP‐immunoreactive neurons, with a mean density of about 29,000 neurons/mm3. No significant sexual dimorphism or age‐related alterations in the population of VP neurons is found. The VIP positive neurons are mainly located in the ventral and central part of the SCN and extend rostrocaudally in a similar way in females and males (1.07 ± 0.08 mm and 1.02 ± 0.11 mm, respectively). The volume of the VIP subdivision is 0.034 ± 0.004 mm3 and contains 1,700 ± 140 VIP‐immunoreactive neurons, with a mean density of about 63,000 neurons/mm3. An age‐dependent sexual dimorphism is observed in the number of VIP‐expressing neurons in the SCN: young males have about twice as many VIP neurons as females of the same age, whereas in middle‐aged subjects this sexual difference is reversed, and less robust, with females now having about 1.7 times as many VIP neurons as males. In old subjects the difference in VIP cell number between men and women disappears. Conclusions The present study clearly shows that the population of VP neurons in the human SCN is considerably larger than the population of VIP neurons. Furthermore, the age‐related sexual differences in the VIP cell number reinforces the idea that the SCN is not only involved in the timing of circadian rhythms but also in the temporal organization of reproductive functions.