Supervisor: prof. dr hab. Zuzanna Setkowicz-Janeczko (zuzanna.setkowicz@uj.edu.pl)

Institute of Zoology and Biomedical Research

Background information:

Up to 30% of clinical cases of epilepsy are drug-resistant. Among them, there are also women of reproductive age and sometimes even in the period of pregnancy. An alternative to pharmacological treatment may be a ketogenic diet (KD). KD is a special type of diet that mimics a fasting state where the source of energy are fats. Few studies describe its effects on pregnant women suffering from epilepsy or diabetes, ignoring possible long-term changes in their offspring. Our previous studies have shown that KD applied to pregnant females delays performance of most neurodevelopmental tests in pups born to these females. At the same time, we have found that switching KD by a standard diet immediately after birth may abolish these negative changes. It is known that the composition of the microbiome is important in maintaining the body homeostasis. In the newborn pups, the microbiome is passed down from the mother even before birth and then reinforced by the presence of the mother's microbiome during feeding period.

The main question to be addressed in the project:

• How KD treatment of rat females during the period from the conception day to the end of lactation or only during pregnancy will affect the development of nervous system of their offspring?
• Is it possible to eliminate the effects of KD used in pregnancy by normal diet introduced on the second day postpartum?
• How does the gut microbiome change during development in the offspring of those mothers who were fed with KD during pregnancy or pregnancy and lactation.

Information on the methods/description of work:

To answer the questions we propose to examine brains of offsprings from females fed with KD or a standard diet during pregnancy or pregnancy and lactation period. The brains will be collected at 2, 6, 14, 30 or 60 days postpartum together with samples of gut microbiome. In addition, 30 and 60 day-old offspring of these females will be subjected to behavioral tests of memory and anxiety level to check whether any KD-caused deficits can be abolished after replacing KD with normal diet which, consequently, will modify the gut microbiome. In addition, ex vivo MRI imaging will be performed to reveal possible changes in the structure of the brain and its internal connections (tractography). Histological examinations of the brains will also be performed to assess changes in glial and neural components.

Additional information: (eg. Special requirements for the student)

Knowledge of experimental methods that will be used in the project. The ability to animal care during procedures. The ability to perform statistical calculations for presenting results at conferences. Involvement in the preparation of a doctoral dissertation.

References:

[1] Eadie M.J. 2012. Shortcomings in the current treatment of epilepsy. Expert Rev Neurother. 12: 1419-27. doi: 10.1586/ern.12.129

[2] Kosiek et al. 2022. Ketogenic diet impairs neurological development of neonatal rats and affects biochemical composition of maternal brains: evidence of functional recovery in pups. Brain Struct Funct. 227: 1099-1113. doi: 10.1007/s00429-021-02450-1

[3] Włodarek et al. 2019. Role of Ketogenic Diets in Neurodegenerative Diseases (Alzheimer's Disease and Parkinson's Disease). Nutrients. 11 :169. doi: 10.3390/nu11010169