Multimodal analysis reveals cellular diversity and divergent circuits of the zona incerta

R Ryan Kast (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) S Sean K. Simmons (Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard) N Nita Rome (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) X Xian Adiconis (Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard) L Lisa Melamed (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) K Kathryn A. McCabe (McGovern Institute for Brain Research, Massachusetts Institute of Technology) D Dongqing Wang (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) A Alexandra Krol (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) M Magdalena Krubner (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) V Vincent Prevosto (McGovern Institute for Brain Research, Massachusetts Institute of Technology) K Kirsten Levandowski (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) U Ubadah Sabbagh (Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology) H Heather A. Sullivan (McGovern Institute for Brain Research, Massachusetts Institute of Technology) I Ian R. Wickersham (McGovern Institute for Brain Research, Massachusetts Institute of Technology) F Fan Wang J Joshua Z. Levin (Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard) Z Zhanyan Fu (McGovern Institute for Brain Research, Massachusetts Institute of Technology) G Guoping Feng

Abstract

The zona incerta (ZI) plays an important role in diverse behavioral functions and is an emerging clinical target for deep brain stimulation to treat neurological conditions. Despite their importance, the cell type composition of the ZI and the anatomical circuit organization linking specific ZI cell types to brain-wide circuits remain unclear. In this study, we used single-nucleus RNA-sequencing, spatial RNA profiling, ex vivo electrophysiology, and anatomical circuit mapping to generate a multimodal cellular atlas of the ZI and to characterize the brain-wide monosynaptic inputs to specific ZI cell types. We define four genetically distinct populations of ZI GABAergic neurons that display unique spatial distributions, specialized intrinsic electrophysiological properties, divergent efferent projections, and cell-type-dependent synaptic input patterns. Finally, using fiber photometry, we reveal response divergence between distinct ZI cell types in a behavioral assay that centers around approach-avoidance conflict produced by appetitive and innately aversive stimuli. This study thus provides a foundational resource for the design and interpretation of future experiments aimed at understanding the organization and function of the ZI.

Article Details

Volume / Issue Vol. 123, Issue 18
Published May 05, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (18)

R

Ryan Kast

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

S

Sean K. Simmons

Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard

N

Nita Rome

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

X

Xian Adiconis

Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard

L

Lisa Melamed

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

K

Kathryn A. McCabe

McGovern Institute for Brain Research, Massachusetts Institute of Technology

D

Dongqing Wang

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

A

Alexandra Krol

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

M

Magdalena Krubner

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

V

Vincent Prevosto

McGovern Institute for Brain Research, Massachusetts Institute of Technology

K

Kirsten Levandowski

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

U

Ubadah Sabbagh

Yang Tan Collective, Hock E. Tan and K. Lisa Yang Center for Autism Research at MIT, Massachusetts Institute of Technology

H

Heather A. Sullivan

McGovern Institute for Brain Research, Massachusetts Institute of Technology

I

Ian R. Wickersham

McGovern Institute for Brain Research, Massachusetts Institute of Technology

F

Fan Wang

J

Joshua Z. Levin

Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard

Z

Zhanyan Fu

McGovern Institute for Brain Research, Massachusetts Institute of Technology

G

Guoping Feng