Prof. Ingrid Nylander

Radioimmunoassay on peptides in heat stabilized brain tissue 

Professor Ingrid Nylander has always been interested in pharmacology; how substances affect the human body and behavior. She studied pharmacology at Uppsala University, and joined Professor Lars Terenius group after her studies to pursue a PhD. During her PhD work Professor Nylander came in contact with opioids and peptides and developed a radioimmunoassay (RIA) protocol for detecting dynorphin in brain samples.

Interest in the role of peptides in addiction

After her PhD work was completed, Ingrid Nylander went to Louisiana State University for a post doc period, where the focus was behavioral studies of drug addiction. The main scope of the project was to study receptivity of cocaine and heroin, but also endogenous opioid peptides. Keeping peptides intact has always been tricky, why much of the latter studies were unsuccessful and, in hindsight, impossible from the start. After her post-doc in the U.S., professor Nylander returned to Sweden, but this time to the Karolinska Institute. Here she continued to explore addiction, primarily alcohol addiction, in animal models and tried to sort out how peptides and neurotransmittors regulate each other. Professor Nylander wanted to combine behavioral studies with neurochemistry, and hence quantitative studies of neuropeptide-levels were an important part of her research. At the Karolinska Institute Professor Nylander had access to a focused microwave applicator, where heat is used to stabilize peptides and proteins in the rodent brain; she even published a paper already in 1997, describing the importance of preserving your tissue sample to ensure correct levels of peptides in your results.

A professor interested in effects of early life stress

In 2005 Ingrid Nylander was appointed full professor at Uppsala University and now heads the “Neuropharmacology, addiction & behavior”-group at the department of Pharmaceutical Biosciences. She has developed a model for investigation of consequences of early-life stress by using maternal separation attempting to answer the fundamental question: why do some individuals develop addiction and how does childhood trauma contribute to this risk? In the rat model, the mother is separated from its pups for long periods of time, up to 6 hours, daily during the three first weeks of life. This separation has been shown to cause stress to the pups and permanent disturbances in the opioid system. The effect is consistent throughout adulthood; i.e. vulnerability and addiction are clearly affected by negative early-life events and the biological changes caused in the developing brain are a contributing factor.

Combining behavioral studies with neurobiology – how to measure the impact of trauma

After behavioral studies, the rats are sacrificed and the brain tissue is analyzed regarding receptors with receptor binding studies or autoradiography, enzymes with enzyme activity assays and peptides using RIA. As peptides are hard to preserve, Professor Nylander was curious what heat stabilization could do for her samples. In her previous protocol, samples were kept on ice until boiled in acetic acid, but still degradation occurred. With heat stabilization, not only is the peptide stability assured, but also sample handling becomes much easier. With the heat stabilized samples Professor Nylander feels secure that she measures as correct levels as possible.