Serotonin regulation of behaviour via large-scale neuromodulation of serotonin receptor networks

Although we understand how serotonin receptors function at the single-cell level, what role different serotonin receptors play in regulating brain-wide activity and, in turn, human behaviour, remains unknown. Here, we developed transcriptomic-neuroimaging mapping to characterise brain-wide functional signatures associated with specific serotonin receptors: serotonin receptor networks (SRNs). Probing SRNs with optogenetics-fMRI and pharmacology in mice, we show that activation of dorsal raphe serotonin neurons differentially modulates the amplitude and functional connectivity of different SRNs, showing that receptors’ spatial distributions can confer specificity not only at the local, but also at the brain-wide, network-level. In humans, using resting state fMRI, different sets of SRNs are linked to different behavioural phenotypes. These results provide compelling evidence that heterogeneous brain-wide distributions of different serotonin receptor types may underpin behaviourally-distinct modes of serotonin regulation. This suggests that dorsal raphe serotonin neurons may regulate multiple aspects of human behaviour via modulation of large-scale receptor networks.

Developmental Vitamin D Deficiency in Pregnant Rats Does Not Induce Preeclampsia

Preeclampsia is a pregnancy disorder characterized by hypertension. Epidemiological studies have associated preeclampsia with an increased risk of neurodevelopmental disorders in offspring, such as autism and schizophrenia. Preeclampsia has also been linked with maternal vitamin D deficiency, another candidate risk factor also associated with autism. Our laboratory has established a gestational vitamin-D-deficient rat model that shows consistent and robust behavioural phenotypes associated with autism- and schizophrenia-related animal models. Therefore, we explored here whether this model also produces preeclampsia as a possible mediator of behavioural phenotypes in offspring. We showed that gestational vitamin D deficiency was not associated with maternal blood pressure or proteinuria during late gestation. Maternal and placental angiogenic and vasculogenic factors were also not affected by a vitamin-D-deficient diet. We further showed that exposure to low vitamin D levels did not expose the placenta to oxidative stress. Overall, gestational vitamin D deficiency in our rat model was not associated with preeclampsia-related features, suggesting that well-described behavioural phenotypes in offspring born to vitamin-D-deficient rat dams are unlikely to be mediated via a preeclampsia-related mechanism.

A brain-specific pgc1α fusion transcript affects gene expression and behavioural outcomes in mice

PGC1α is a transcriptional coactivator in peripheral tissues, but its function in the brain remains poorly understood. Various brain-specific Pgc1α isoforms have been reported in mice and humans, including two fusion transcripts (FTs) with non-coding repetitive sequences, but their function is unknown. The FTs initiate at a simple sequence repeat locus ∼570 Kb upstream from the reference promoter; one also includes a portion of a short interspersed nuclear element (SINE). Using publicly available genomics data, here we show that the SINE FT is the predominant form of Pgc1α in neurons. Furthermore, mutation of the SINE in mice leads to altered behavioural phenotypes and significant up-regulation of genes in the female, but not male, cerebellum. Surprisingly, these genes are largely involved in neurotransmission, having poor association with the classical mitochondrial or antioxidant programs. These data expand our knowledge on the role of Pgc1α in neuronal physiology and suggest that different isoforms may have distinct functions. They also highlight the need for further studies before modulating levels of Pgc1α in the brain for therapeutic purposes.

Behavioural and molecular characterisation of the Dlg2 haploinsufficiency rat model of genetic risk for psychiatric disorder

Genetic studies implicate disruption to the DLG2 gene in copy number variants as increasing risk for schizophrenia, autism spectrum disorders and intellectual disability. To investigate psychiatric endophenotypes associated with DLG2 haploinsufficiency (and concomitant PSD-93 protein reduction) a novel clinically relevant Dlg2+/- rat was assessed for abnormalities in anxiety, sensorimotor gating, hedonic reactions, social behaviour, and locomotor response to the N-Methyl-D-aspartic acid receptor antagonist phencyclidine. Dlg gene and protein expression were also investigated to assess model validity. Reductions in PSD-93 messenger RNA and protein were observed in the absence of compensation by other related genes or proteins. Behaviourally Dlg2+/- rats show potentiated locomotor response to phencyclidine, as is typical of psychotic disorder models, in the absence of deficits in the other behavioural phenotypes assessed here. This shows that the behavioural effects of Dlg2 haploinsufficiency may specifically relate to psychosis vulnerability but are subtle, providing a contrast to the gross deficits in Dlg2 homozygous models (Winkler, et al., 2018; Yoo et al., 2020a) which do not so specifically model the single chromosome DLG2 deletion in carriers of risk-associated copy number variants.

Proteomic Analysis of Hydromethylthionine in the Line 66 Model of Frontotemporal Dementia Demonstrates Actions on Tau-Dependent and Tau-Independent Networks

Abnormal aggregation of tau is the pathological hallmark of tauopathies including frontotemporal dementia (FTD). We have generated tau-transgenic mice that express the aggregation-prone P301S human tau (line 66). These mice present with early-onset, high tau load in brain and FTD-like behavioural deficiencies. Several of these behavioural phenotypes and tau pathology are reversed by treatment with hydromethylthionine but key pathways underlying these corrections remain elusive. In two proteomic experiments, line 66 mice were compared with wild-type mice and then vehicle and hydromethylthionine treatments of line 66 mice were compared. The brain proteome was investigated using two-dimensional electrophoresis and mass spectrometry to identify protein networks and pathways that were altered due to tau overexpression or modified by hydromethylthionine treatment. Overexpression of mutant tau induced metabolic/mitochondrial dysfunction, changes in synaptic transmission and in stress responses, and these functions were recovered by hydromethylthionine. Other pathways, such as NRF2, oxidative phosphorylation and protein ubiquitination were activated by hydromethylthionine, presumably independent of its function as a tau aggregation inhibitor. Our results suggest that hydromethylthionine recovers cellular activity in both a tau-dependent and a tau-independent fashion that could lead to a wide-spread improvement of homeostatic function in the FTD brain.

Effects of multigenerational exposure and phenotypic variation on a freshwater fish species exposed to elevated carbon dioxide (CO2)

The amount of dissolved carbon dioxide (CO2) and the acidity of aquatic ecosystems is increasing as atmospheric CO2 concentrations increase due to human activities. Changes in pH and dissolved CO2 can have considerable aversive effects on fish physiology and behaviour, which can result in negative effects on fish populations. Multigenerational studies have found that the conditions experienced by parents can have significant effects on the performance of their offspring and understanding these effects can help to predict how fish populations will cope in future conditions. Additionally, repeatable behavioural phenotypes are good predictors of trends in behaviour, can be useful predictors of other physiological and life history traits, and can be subject to selection pressures. Unfortunately, the effects of elevated CO2 on freshwater fishes over multiple generations, and the effects of behavioural phenotypes, are poorly understood. In my thesis, freshwater Japanese Medaka (Oryzias latipes) were used to investigate the influence of phenotypic variation and differences in time of exposure (generational) on biological responses to elevated CO2. Lab-reared medaka were divided into ‘responsive’ and ‘non-responsive’ groups based on behavioural differences from the population mean during acute exposure to high CO2 in a common shuttling and novel tank behavioural assay. Responsive and non-responsive fish in parental generation (P) were subdivided and exposed to either control (~480 ppm) or high CO2 (~1250 ppm) conditions over a 6-week period. Following this time, eggs from this generation were collected and randomly selected into either high or control conditions, where they were hatched and reared until maturation (filial generation one (F1), 18 weeks). Eggs from F1 were collected and hatched and reared in the same conditions as their parents until adulthood (filial generation two (F2), 24 weeks). Body condition (size, weight and length), behaviour (total distance moved, time spent in the outer zone of the behavioural arena, and swimming direction), reproductive (number of eggs, size of eggs, and survival to hatch) performance, and the relative abundance of various mRNA transcripts in whole brain tissue of fish was measured across these three generations. Behavioural phenotypes influenced reproduction for P and F2 generation fish, and growth for F1 and F2 fish; suggesting that intraspecific variation in behavioural phenotypes may influence how medaka respond to elevated CO2. However, behavioural phenotypes did not have a significant effect on mRNA abundance on genes targeted in my study. Multigenerational exposure to elevated CO2 were shown to improve the performance of offspring in some measures and resulted in changes of mRNA abundance of several genes. Transgenerational exposure, where a parent or grandparent was exposed to elevated CO2 but the offspring were not exposed to elevated CO2, resulted in some deleterious effects suggesting that, generally, exposure to environmental conditions that differ from that of their parents may put fish especially at risk. In my thesis, current CO2 exposure appeared to be the best predictor of overall condition, where fish exposed to elevated CO2 were worse off than fish exposed to control CO2 conditions. The results of this research contribute to filling a current gap of knowledge in understanding how freshwater fish will respond to future conditions over an ecologically-relevant time scale. Importantly, this information will contribute to generating more informed decisions on freshwater ecosystem management and future research directions. Marine and freshwater environments offer food and water security and are of high importance to the economy and the health of our planet, making my research relevant to our broader society.