Summary: Study identifies rare brain cell types that are unique to male mice, and others that are unique to females. The cells were discovered in the hypothalamus, a region of the brain that governs both aggression and mating behaviors.
Caltech researchers have discovered rare brain cell types that are unique to male mice and other types that are unique to female mice. These sex-specific cells were found in a region of the brain that governs both aggression and mating behaviors.
The study was done as a collaboration between the laboratory of David Anderson, Seymour Benzer Professor of Biology, Tianqiao and Chrissy Chen Institute for Neuroscience Leadership Chair, Howard Hughes Medical Institute Investigator, and director of the Tianqiao and Chrissy Chen Institute for Neuroscience; and a team at the Allen Institute for Brain Sciences in Seattle, Washington. A paper describing the research appears online on October 17 in the journal Cell.
“The results show that there are differences between male and female mammalian brains at the level of cellular composition as well as gene expression but that those differences are subtle, and their functional significance remains to be explained,” says Anderson.
There are many different types of cells within the brain, such as neurons that transmit signals and glial cells that support neural functions. Although all of these cells contain the same set of genes, or genome, the types of cells differ in how they express those genes. As an analogy, one can imagine the genome as an 88-key piano in each cell. Each cell does not use all 88 keys. Therefore, the subset of keys that the cell “plays” determines the type of cell it is.
The hypothalamus is a fundamental region of the brain found in all vertebrates including humans. Previous studies have shown that a specific anatomic subdivision in the hypothalamus, called the ventrolateral subdivision of the ventromedial hypothalamus (VMHvl), contains cells that control aggression and mating behaviors. In these studies, strong stimulation of these neurons in male and female mice immediately caused the animals to become aggressive, even in the absence of any threat. However, weak stimulation caused the mice to begin mating behaviors.
In this new work, led by Caltech graduate student Dong-Wook Kim, the researchers examined gene expression in individual cells in the VMHvl. This was made possible by advanced transcriptomic techniques that can enumerate and identify the RNA transcripts that a cell contains; this information can then be used to classify different cell types. Previous studies were only able to examine 10 percent of the transcripts in each cell, whereas this study looked at a larger proportion of transcripts. The team discovered that there are 17 different types of brain cells in this tiny region alone. What is more, an examination of the patterns of gene expression revealed that some of these 17 cell types are much more abundant in male mice than in females, while others are found only in females.
It was known that different genes are expressed in the two mouse sexes–indeed, a genetic test can tell you whether a mouse is male or female–but this is the first discovery of types of cells that are sex-specific in a mammalian brain. Cells are considered to be distinct types when the expression of large clusters of genes varies from cell to cell.
Future work will try to determine the functions of these differing cell types.
The work was a collaboration between several different Caltech laboratories. In addition to Anderson, other Caltech faculty co-authors are Yuki Oka, assistant professor of biology and Chen Scholar; Lior Pachter (BS ’94), Bren Professor of Computational Biology and Computing and Mathematical Sciences; and Long Cai, professor of biology and biological engineering. Cai, Oka, and Pachter are affiliated faculty members of the Tianqiao and Chrissy Chen Institute for Neuroscience at Caltech.
Other Caltech co-authors include former graduate students Lynn Yi (PhD ’19) and Sheel Shah (PhD ’17), research technician Noushin Koulena, research technician assistant Nico Pierson, research specialist Li-Ching Lo, and postdoctoral scholar Allan-Hermann Pool. Additional co-authors include Zizhen Yao, Lucas Graybuck, Tae Kyung Kim, Thuc Nghi Nguyen, Kimberly Smith, Olivia Fong, Bosiljka Tasic, and Hongkui Zeng, all of the Allen Institute for Brain Science.
Funding: Funding was provided by the National Institutes of Health (NIH) BRAIN Initiative, the NIH, and the Howard Hughes Medical Institute.
Multimodal Analysis of Cell Types in a Hypothalamic Node Controlling Social Behavior
Highlights • SMART-seq and 10x platforms reveal 17 transcriptomic cell types (T-types) in VMHvl • Multiplexed smFISH confirms scRNA-seq and identifies T-types with A-P restrictions • Rare female- and male-specific T-types are found among the Esr1 + subclusters • Relatively few VMHvl T-types exhibit behavior-specific activation and connectivity
Summary The ventrolateral subdivision of the ventromedial hypothalamus (VMHvl) contains ∼4,000 neurons that project to multiple targets and control innate social behaviors including aggression and mounting. However, the number of cell types in VMHvl and their relationship to connectivity and behavioral function are unknown. We performed single-cell RNA sequencing using two independent platforms—SMART-seq (∼4,500 neurons) and 10x (∼78,000 neurons)—and investigated correspondence between transcriptomic identity and axonal projections or behavioral activation, respectively. Canonical correlation analysis (CCA) identified 17 transcriptomic types (T-types), including several sexually dimorphic clusters, the majority of which were validated by seqFISH. Immediate early gene analysis identified T-types exhibiting preferential responses to intruder males versus females but only rare examples of behavior-specific activation. Unexpectedly, many VMHvl T-types comprise a mixed population of neurons with different projection target preferences. Overall our analysis revealed that, surprisingly, few VMHvl T-types exhibit a clear correspondence with behavior-specific activation and connectivity.