Original Article

Experimental Neurobiology 2019; 28(6): 709-719

Published online December 31, 2019

© The Korean Society for Brain and Neural Sciences

Three-Dimensional Analysis of Mouse Habenula Subnuclei Reveals Reduced Volume and Gene Expression in the Lipopolysaccharide-mediated Depression Model

Esther Yang, Jin Yong Kim, Soo Hyun Yang, Eunsoo Lee, Woong Sun, Hyun Woo Lee* and Hyun Kim*

Department of Anatomy, College of Medicine, Korea University, Seoul 02841, Korea

Correspondence to: *To whom correspondence should be addressed.
Hyun Woo Lee, TEL: 82-2-2286-1385, FAX: 82-2-929-5696
Hyun Kim, TEL: 82-2-2286-1153, FAX: 82-2-929-5696
These authors contributed equally to this work.

Received: September 25, 2019; Revised: November 25, 2019; Accepted: November 27, 2019

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License
( which permits unrestricted non-commercial use, distribution, and
reproduction in any medium, provided the original work is properly cited.

The habenula (Hb) is small but important brain structure, anatomically and functionally links the forebrain with the midbrain to modulate various neuropsychiatric functions associated with drug addiction and emotion-associated dysfunctions. Several reports suggested that the dysfunction of Hb-related functions affected the Hb structurally and functionally. However, the technical limitation has awaited the solid conclusion of whether Hb change due to depression is likely to occur in certain subnuclei of the Hb. To probe this possibility, we developed 3-dimensional reconstruction methods for the high-resolution volumetric analysis of Hb and the mRNA levels at the given volume in normal or lipopolysaccharide (LPS)-mediated mouse model of depression. Notably, we discovered that the volume reduction was prominent in medial Hb but not in lateral Hb after LPS treatments. On the other hand, the RNA expression levels of known Hb regional markers such as Tac1 (dorsal part of medial Hb), ChAT (ventral part of medial Hb), and Tacr1 (medial and lateral Hb) were all decreased in all Hb subnuclei in LPS-injected mice. Accordingly, accurate volumetry with marker labeling was not feasible. Collectively, these established 3D analyses of mouse Hb successfully and precisely determine the volume-based changes of small brain structure, which should be applicable in a wider range of mouse models or pathological specimens.

Graphical Abstract

Keywords: 3-dimensional structure, Gene expression, Depression, Habenula