Characterization of Microglial Activation in Hypothalamic

Nuclei of Male and Female Mice

Introduction:

Inflammation occurs in the body in response to infection or injury and plays a role in the

pathophysiology of many diseases such as Alzheimer’s disease, diabetes, and some cancers.

(Kirkpatrick & Miller, 2013) Innate inflammation has been linked to both psychiatric disorders

and the glutamatergic and monaminergic abnormalities associated with them (Najjar et al.

2013). Recent data reveals “reliable associations of inflammatory markers with psychiatric

disorders, the induction of psychiatric symptoms following administration of inflammatory

stimuli, the association of inflammation-related genes with psychiatric disease, and the

elucidation of neurobiological and immunological mechanisms by which inflammation targets

neurotransmitters and neuro-circuits to change behavior” (Miller & Raison, 2015). With the

large role that inflammation plays in an assortment of diseases, it is important to look at the

varying ways that inflammation is induced in males and females and the physiological

consequences it has.

Microglia cells are the resident macrophages of the central nervous system, comprising

approximately 12% of the brain (Lull & Block, 2010). In a normal, healthy brain, microglia are

referred to as ‘resting’, and they are distinguished by their morphology (Well, lots of other

things too, but morphology is a big one. Cite this.) In this state, they have the ability scan the

environment by extending and retracting their processes in order to survey the environment

and identify signals that require a response (Tremblay et al. 2011; Lull, Block, 2010). Microglia
become activated after sensing changes in the environment, such as dead or dying cells, foreign

material, or pathogens (citation). In response to these signals, they undergo a drastic change in

morphology, characterized by their amoeboid shape, and can release reactive oxidative species,

pre- or anti- inflammatory factors, factors that recruit other cells, and more (Lull, Block, 2010).

They can also regulate the growth of new neurons, along with their formation, survival, and

elimination (McCarthy & Wright, 2017). Their goal is to return the brain to a state of

homeostasis.

Microglia can differentiate in males in females from a young age due to androgen

production by the fetal testis. The overall brain development and differentiation can cause

sexually dimorphic neuroanatomical changes that have a prominent impact on behavior,

sexually dimorphic meaning that there are two forms, on in males and one in females

(McCarthy, 2016). The importance of studying sex-differences in the brain cannot be

overlooked – the differing physiological changes in the brain between males and females could

play an unprecedented role in the generation of new therapies and procedures for a variety of

brain diseases. One of the areas of the brain that has sexually dimorphic implications is the

preoptic area (POA).

The POA is a region that extends from the posterior edge of the optic chiasm to the

anterior end of the hypothalamus. It has been noted to play a role in the detection of hormones

in the blood, regulates body temperature, cardiovascular function, fluid balance, water intake,

and more (Saper, 2012). It is also notable for its sexually dimorphic nuclei and its robust

neuroanatomical sex differences (McCarthy & Wright, 2017). The differences between males

and females ranged from dendritic organization to the size of sub nuclei, being 3-5 times larger
in males. These differences have behavioral consequences (McCarthy & Wright, 2017). The POA

is responsible for male sexual and parental behaviors, and lesions to this area eliminate both

drives. Microglia play a critical role in creating and establishing the sex-specific patterns found

in the POA (McCarthy & Wright, 2017). They are more highly concentrated and are in a more

activated state in males compared to females. The POA shows a clear connection between

microglia morphology and the behavioral differences and consequences that occur because of

it.

The POA is not the only place in the brain that show difference in immune cells between

males and females. In a rodent model, microglia density was significantly decreased in the

hippocampus and increased in the amygdala in males compared to females (Guneykaya et al.

2018). In a study of the cortex, hippocampus, amygdala, striatum, and cerebellum, there was

no significant difference in soma size between males and females at 3 weeks, but the soma size

was significantly larger in males at 13 weeks (Guneykaya et al. 2018). Throughout the brain,

there are key differences between the structure of microglia between males and females, and

those structural changes may have behavioral implications.

It has been demonstrated that there sexually dimorphic structural differences in the

POA, one nuclei of the hypothalamus. We are interested to see if this is true for other nuclei,

specifically the hypothalamic arcuate nucleus (ARC). The ARC is in an area of the brain that has

access to all of the nutrients and hormones circulating in the blood because of the relatively

permeable blood brain barrier that is associated with it (Roh & Kim, 2016). This allows the ARC

directly survey the substances that are entering the brain. The ARC has the ability to mediate

leptin, a protein that plays a role in the storage of fat in the body, and in locomotion activity
(Roh & Kim, 2016). This region is essential for the initiation of signals that cause us to change

behaviors in order to survive, like eating, drinking, and more. Specifically, we are looking at this

area because of its close proximity to proopiomelanocortin (POMC) neurons. It has previously

been demonstrated that there are physiological differences between the way microglia are

expressed in men and women in some parts of the brain, we are interested in determining if

microglial variations in this area have any consequences on drinking behavior.

POMC neurons originate in the ARC and extend all along the rostroaudal axis of the

brain, but its integration through the central nervous system (CNS) remains unclear (Mercer et

al. 2013). Beta-endorphins, one of the three primary opioids of the CNS, are a downstream

peptide product of the POMC gene (Mercer et al. 2013). These endorphins play a role in food

reward circuitry, and influence body mass, food intake, and metabolic output (Mercer et al.

2013).

*Significance* - “Being male imparts a major risk for the development of a developmental

neurological or neuropsychiatric disorder, whereas being female appears to afford some

protection from the same. Identifying the biological origins of male vulnerability and female

protection will generate novel targets for therapeutic intervention and prevention.” -

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5286722/
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Transcriptional and Translational Differences of Migroglia from Male and Female Brains.
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Kirkpatrick, B., Miller, B. (2013). Inflammation and Schizophrenia, Schizophrenia Bulletin,

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Miller, A. H., & Raison, C. L. (2015). Are Anti-inflammatory Therapies Viable Treatments for
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Mercer, A. J., Hentges, S. T., Meshul, C. K., & Low, M. J. (2013). Unraveling the central
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