A Novel Method to Estimate Levels of Receptors Using an Allosteric Modulator: Focus on Muscarinic M1 Receptor.

This chapter outlines some of the general principles that need to be considered when developing a radioligand binding assay to measure the affinity and density of radioligand binding to a receptor in tissue or on cells. In addition it describes an innovative step forward in using radioligand binding assays to measure levels of muscarinic M1 receptors in human postmortem CNS, using both membrane binding and in situ radioligand binding. These examples show how, using receptor-specific allosteric modulators, it is possible to gain an estimate of the density of a single receptor using a radioligand that is not totally specific to the target site of interest. Given there is a growing understanding that there are problems with antibodies not showing specificity to their supposed target protein, well-characterized radioligand binding techniques still provide an important tool when studying receptor density in tissues and cells.

Evidence that the frontal pole has a significant role in the pathophysiology of schizophrenia.

Different regions of the cortex have been implicated in the pathophysiology of schizophrenia. Recently published data suggested there are many more changes in gene expression in the frontal pole (Brodmann's Area (BA) 10) compared to the dorsolateral prefrontal cortex (BA 9) and the anterior cingulate cortex (BA 33) from patients with schizophrenia. These data argued that the frontal pole is significantly affected by the pathophysiology of schizophrenia. The frontal pole is a region necessary for higher cognitive functions and is highly interconnected with many other brain regions. In this review we summarise the growing body of evidence to support the hypothesis that a dysfunctional frontal pole, due at least in part to its widespread effects on brain function, is making an important contribution to the pathophysiology of schizophrenia. We detail the many structural, cellular and molecular abnormalities in the frontal pole from people with schizophrenia and present findings that argue the symptoms of schizophrenia are closely linked to dysfunction in this critical brain region.

Lower levels of tubulin alpha 1b in the frontal pole in schizophrenia supports a role for changed cytoskeletal dynamics in the aetiology of the disorder.

Our transcriptomic study suggested there were markedly lower levels of tubulin alpha 1b (TUBA1B) expression in BA 10, but not BA 9, from patients with schizophrenia. We now use Western blotting to compare levels of TUBA1B protein in BA 9 and 10 from patients with schizophrenia and BA 10 from patients with mood disorders to controls as well as in the frontal cortex from rats after treatment with haloperidol, chlorpromazine or vehicle for 28 days. Levels of TUBA1B were significantly lower (- 18.6%) in BA 10, but not BA 9, from patients with schizophrenia. Levels of TUBA1B did not differ significantly from controls in BA 10 from patients with mood disorders or in the cortex of rats after antipsychotic drug treatments. Levels of TUBA1B were significantly lower (- 30%) in BA 10 from patients with schizophrenia who were not being treated with antipsychotic drugs close to death compared to those who were treated close to death. These data suggest that lower levels of TUBA1B, a cytoskeletal protein, in BA 10 from patients with schizophrenia are not a simple drug effect and therefore add to the hypothesis that a breakdown in cytoskeletal homoeostasis may be contributing to the genesis of the symptoms of the disorder.