Long-Term Consumption of High-Fat Diet in Rats: Effects on Microglial and Astrocytic Morphology and Neuronal Nitric Oxide Synthase Expression

2016-08-15 20:04:03

Cellular and Molecular Neurobilogy; 15 August 2016: DOI: 10.1007/s10571-016-0417-5

Kinga Gzielo, Michal Kielbinski, Jakub Ploszaj, Krzysztof Janeczko, Stefan P. Gazdzinski, Zuzanna Setkowicz


Obesity in humans is associated with cognitive decline and elevated risk of neurodegenerative diseases of old age. Variations of high-fat diet are often used to model these effects in animal studies. However, we previously reported improvements in markers of memory and learning, as well as larger hippocampi and higher metabolite concentrations in Wistar rats fed high-fat, high-carbohydrate diet (HFCD, 60 % energy from fat, 28 % from carbohydrates) for 1 year; this diet leads to mild ketonemia (Setkowicz et al. in PLoS One 10:e0139987, 2015). In the present study, we follow up on this cohort to assess glial morphology and expression of markers related to gliosis. Twenty-five male Wistar rats were kept on HFCD and twenty-five on normal chow. At 12 months of age, the animals were sacrificed and processed for immunohistochemical staining for astrocytic (glial fibrillary acidic protein), microglial (Iba1), and neuronal (neuronal nitric oxide synthetase, nNOS) markers in the hippocampus. We have found changes in immunopositive area fraction and cellular complexity, as studied by a simplified Sholl procedure. To our knowledge, this study is the first to apply this methodology to the study of glial cells in HFCD animals. GFAP and Iba1 immunoreactive area fraction in the hippocampi of HFCD-fed rats were decreased, while the mean number of intersections (an indirect measure of cell complexity) was decreased in GFAP-positive astrocytes, but not in Iba1-expressing microglia. At the same time, nNOS expression was lowered after HFCD in both the cortex and the hippocampus.


In 2014, more than 1.9 billion adults, 18 years and older, were overweight. Of these, over 600 million were obese. Once considered a problem only in high-income countries, obesity has reached epidemic proportions also in low- and middle-income countries. In the nervous system, obesity has been linked to increased oxidative stress and inflammation, particularly involving astrocytes and microglia. In the end, obesity results in cognitive decline in humans and impairment in different tasks in animals. Multiple different approaches to modeling obesity in experimental animals exist. Variants of high fat, or high-fat and high-carbohydrate (HFCD) diet are experimental tools often used to model the effects of energy dense food. Of special interest are the effects of such a diet on hippocampal function, as numerous rodent studies indicate that highly caloric diets impair function and structure of the hippocampus, leading to alterations in hippocampus-dependent long-term spatial memory. There is also mounting evidence that increased inflammation, oxidative stress and altered energy metabolism linked to HFCD consumption are all tied to mechanisms of cognitive decline relevant both to Alzheimer’s disease and normal aging. On the other hand, the relationship between dietary fat and caloric intake, age, and cognitive decline is still controversial, with some authors finding either no negative or weakly positive effects of high-calory diets. For this reason, it is important to further investigate the influence of high-fat and high-carbohydrate diets on brain function at different points during lifespan, especially in mature and aging animals.

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Key Words

High-fat diet | nNOS | Astrocyte | Microglia | Hippocampus | Sholl analysis