A retaining sphenoid and dura procedure in the rat to obtain intact pituitary-infundibulum-hypothalamus preparations

https://doi.org/10.1016/j.jneumeth.2020.108694Get rights and content
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Highlights

  • We present a procedure to obtain intact rat hypothalamus-pituitary brain specimen.

  • The present method protects the integrity of the fragile infundibulum (pituitary stalk).

  • Our method is based on the anatomy of the rat head, skull, brain and meninges.

  • Examples are illustrated of histology and 3D tissue clearing in the hypothalamic-pituitary continuity.

Abstract

Background

The histopathological study of brain tissue is a conventional method in neuroscience. However, procedures specifically developed to recover intact hypothalamic-pituitary brain specimens, are not available.

New method

We describe a detailed protocol for obtaining intact rat brain with pituitary-hypothalamus continuity through an intact infundibulum. The brain is collected via a ventral approach through removing the skull base. Membranous structures surrounding the hypothalamus-pituitary system can be preserved, including vasculature.

Results

We report a retaining sphenoid and dura technique to obtain intact hypothalamic-pituitary brain preparations, and we confirm the practicability of this method. By combination of this technique with histological analysis or 3D brain tissue clearing and imaging methods, the functional morphology structure of the hypothalamus-pituitary can be further explored.

Comparison with existing method

The current procedure is limited in showing the connection between the hypothalamus and the pituitary. Our procedure effectively protects the integrity of the fragile infundibulum and thus prevents the pituitary from separating from the hypothalamus.

Conclusions

We present a convenient and practical approach to obtain intact hypothalamus-pituitary brain specimens for subsequent histopathological evaluation.

Keywords

Hypothalamohypophyseal system
Pituitary stalk
Rat brain
Arginine vasopressin
Histopathology

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