An adeno-associated viral vector transduces the rat hypothalamus and amygdala more efficient than a lentiviral vector

<p>Abstract</p> <p>Background</p> <p>This study compared the transduction efficiencies of an adeno-associated viral (AAV) vector, which was pseudotyped with an AAV1 capsid and encoded the green fluorescent protein (GFP), with a lentiviral (LV) vector, which was pseudotyped with a VSV-G envelop and encoded the discosoma red fluorescent protein (dsRed), to investigate which viral vector transduced the lateral hypothalamus or the amygdala more efficiently. The LV-dsRed and AAV1-GFP vector were mixed and injected into the lateral hypothalamus or into the amygdala of adult rats. The titers that were injected were 1 × 10⁸or 1 × 10⁹genomic copies of AAV1-GFP and 1 × 10⁵transducing units of LV-dsRed.</p> <p>Results</p> <p>Immunostaining for GFP and dsRed showed that AAV1-GFP transduced significantly more cells than LV-dsRed in both the lateral hypothalamus and the amygdala. In addition, the number of LV particles that were injected can not easily be increased, while the number of AAV1 particles can be increased easily with a factor 100 to 1000. Both viral vectors appear to predominantly transduce neurons.</p> <p>Conclusions</p> <p>This study showed that AAV1 vectors are better tools to overexpress or knockdown genes in the lateral hypothalamus and amygdala of adult rats, since more cells can be transduced with AAV1 than with LV vectors and the titer of AAV1 vectors can easily be increased to transduce the area of interest.</p

Neuropeptide delivery to the brain: a von Willebrand factor signal peptide to direct neuropeptide secretion

<p>Abstract</p> <p>Background</p> <p>Multiple neuropeptides, sometimes with opposing functions, can be produced from one precursor gene. To study the roles of the different neuropeptides encoded by one large precursor we developed a method to overexpress minigenes and establish local secretion.</p> <p>Results</p> <p>We fused the signal peptide from the Von Willebrand Factor (VWF) to a furin site followed by a processed form of the Agouti related protein (AgRP), AgRP_83-132or α-melanocyte stimulating hormone. <it>In vitro</it>, these minigenes were secreted and biologically active. Additionally, the proteins of the minigenes were not transported into projections of primary neurons, thereby ensuring local release. <it>In vivo </it>administration of VWF-AgRP_83-132, using an adeno-associated viral vector as a delivery vehicle, into the paraventricular hypothalamus increased body weight and food intake of these rats compared to rats which received a control vector.</p> <p>Conclusions</p> <p>This study demonstrated that removal of the N-terminal part of full length AgRP and addition of a VWF signal peptide is a successful strategy to deliver neuropeptide minigenes to the brain and establish local neuropeptide secretion.</p

Lentivirus-mediated transgene delivery to the hippocampus reveals sub-field specific differences in expression

<p>Abstract</p> <p>Background</p> <p>In the adult hippocampus, the granule cell layer of the dentate gyrus is a heterogeneous structure formed by neurons of different ages, morphologies and electrophysiological properties. Retroviral vectors have been extensively used to transduce cells of the granule cell layer and study their inherent properties in an intact brain environment. In addition, lentivirus-based vectors have been used to deliver transgenes to replicative and non-replicative cells as well, such as post mitotic neurons of the CNS. However, only few studies have been dedicated to address the applicability of these widespread used vectors to hippocampal cells in vivo. Therefore, the aim of this study was to extensively characterize the cell types that are effectively transduced in vivo by VSVg-pseudotyped lentivirus-based vectors in the hippocampus dentate gyrus.</p> <p>Results</p> <p>In the present study we used Vesicular Stomatitis Virus G glycoprotein-pseudotyped lentivirual vectors to express EGFP from three different promoters in the mouse hippocampus. In contrast to lentiviral transduction of pyramidal cells in CA1, we identified sub-region specific differences in transgene expression in the granule cell layer of the dentate gyrus. Furthermore, we characterized the cell types transduced by these lentiviral vectors, showing that they target primarily neuronal progenitor cells and immature neurons present in the sub-granular zone and more immature layers of the granule cell layer.</p> <p>Conclusion</p> <p>Our observations suggest the existence of intrinsic differences in the permissiveness to lentiviral transduction among various hippocampal cell types. In particular, we show for the first time that mature neurons of the granule cell layer do not express lentivirus-delivered transgenes, despite successful expression in other hippocampal cell types. Therefore, amongst hippocampal granule cells, only adult-generated neurons are target for lentivirus-mediated transgene delivery. These properties make lentiviral vectors excellent systems for overexpression or knockdown of genes in neuronal progenitor cells, immature neurons and adult-generated neurons of the mouse hippocampus in vivo.</p

The melanocortin pathway and energy homeostasis: From discovery to obesity therapy.

BACKGROUND: Over the past 20 years, insights from human and mouse genetics have illuminated the central role of the brain leptin-melanocortin pathway in controlling mammalian food intake, with genetic disruption resulting in extreme obesity, and more subtle polymorphic variations influencing the population distribution of body weight. At the end of 2020, the U.S. Food and Drug Administration (FDA) approved setmelanotide, a melanocortin 4 receptor agonist, for use in individuals with severe obesity due to either pro-opiomelanocortin (POMC), proprotein convertase subtilisin/kexin type 1 (PCSK1), or leptin receptor (LEPR) deficiency. SCOPE OF REVIEW: Herein, we chart the melanocortin pathway's history, explore its pharmacology, genetics, and physiology, and describe how a neuropeptidergic circuit became an important druggable obesity target. MAJOR CONCLUSIONS: Unravelling the genetics of the subset of severe obesity has revealed the importance of the melanocortin pathway in appetitive control; coupling this with studying the molecular pharmacology of compounds that bind melanocortin receptors has brought a new obesity drug to the market. This process provides a drug discovery template for complex disorders, which for setmelanotide took 25 years to transform from a single gene into an approved drug

Genomic Relationships, Novel Loci, and Pleiotropic Mechanisms across Eight Psychiatric Disorders

Genetic influences on psychiatric disorders transcend diagnostic boundaries, suggesting substantial pleiotropy of contributing loci. However, the nature and mechanisms of these pleiotropic effects remain unclear. We performed analyses of 232,964 cases and 494,162 controls from genome-wide studies of anorexia nervosa, attention-deficit/hyper-activity disorder, autism spectrum disorder, bipolar disorder, major depression, obsessive-compulsive disorder, schizophrenia, and Tourette syndrome. Genetic correlation analyses revealed a meaningful structure within the eight disorders, identifying three groups of inter-related disorders. Meta-analysis across these eight disorders detected 109 loci associated with at least two psychiatric disorders, including 23 loci with pleiotropic effects on four or more disorders and 11 loci with antagonistic effects on multiple disorders. The pleiotropic loci are located within genes that show heightened expression in the brain throughout the lifespan, beginning prenatally in the second trimester, and play prominent roles in neurodevelopmental processes. These findings have important implications for psychiatric nosology, drug development, and risk prediction.Peer reviewe

Chemogenetic Activation of Midbrain Dopamine Neurons Affects Attention, but not Impulsivity, in the Five-Choice Serial Reaction Time Task in Rats

Attentional impairments and exaggerated impulsivity are key features of psychiatric disorders, such as attention-deficit/hyperactivity disorder, schizophrenia, and addiction. These deficits in attentional performance and impulsive behaviors have been associated with aberrant dopamine (DA) signaling, but it remains unknown whether these deficits result from enhanced DA neuronal activity in the midbrain. Here, we took a novel approach by testing the impact of chemogenetically activating DA neurons in the ventral tegmental area (VTA) or substantia nigra pars compacta (SNc) on attention and impulsivity in the five-choice serial reaction time task (5-CSRTT) in rats. We found that activation of DA neurons in both the VTA and SNc impaired attention by increasing trial omissions. In addition, SNc DA neuron activation decreased attentional accuracy. Surprisingly, enhanced DA neuron activity did not affect impulsive action in this task. These results show that enhanced midbrain DA neuronal activity induces deficits in attentional performance, but not impulsivity. Furthermore, DA neurons in the VTA and SNc have different roles in regulating attention. These findings contribute to our understanding of the neural substrates underlying attention deficits and impulsivity, and provide valuable insights to improve treatment of these symptoms

Developmental differences in the brain response to unhealthy food cues : An fMRI study of children and adults

BACKGROUND: Food cues are omnipresent and may trigger overconsumption. In the past 2 decades, the prevalence of childhood obesity has increased dramatically. Because children's brains are still developing, especially in areas important for inhibition, children may be more susceptible than adults to tempting food cues. OBJECTIVE: We examined potential developmental differences in children's and adults' responses to food cues to determine how these responses relate to weight status. DESIGN: We included 27 children aged 10-12 y and 32 adults aged 32-52 y. Functional magnetic resonance imaging data were acquired during a food-viewing task in which unhealthy and healthy food pictures were presented. RESULTS: Children had a stronger activation in the left precentral gyrus than did adults in response to unhealthy compared with healthy foods. In children, unhealthy foods elicited stronger activation in the right inferior temporal and middle occipital gyri, left precentral gyrus, bilateral opercular part of the inferior frontal gyrus, left hippocampus, and left middle frontal gyrus. Adults had stronger activation in the bilateral middle occipital gyrus and the right calcarine sulcus for unhealthy compared with healthy foods. Children with a higher body mass index (BMI) had lower activation in the bilateral dorsolateral prefrontal cortex while viewing unhealthy compared with healthy foods. In adults there was no correlation between BMI and neural response to unhealthy compared with healthy foods. CONCLUSIONS: Unhealthy foods might elicit more attention both in children and in adults. Children had stronger activation while viewing unhealthy compared with healthy foods in areas involved in reward, motivation, and memory. Furthermore, children activated a motivation and reward area located in the motor cortex more strongly than adults did in response to unhealthy foods. Finally, children with a higher BMI had less activation in inhibitory areas in response to unhealthy foods, which may mean they are more susceptible to tempting food cues. This trial was registered at www.trialregister.nl as NTR4255

Developmental differences in the brain response to unhealthy food cues : An fMRI study of children and adults

Background: Food cues are omnipresent and may trigger overconsumption. In the past 2 decades, the prevalence of childhood obesity has increased dramatically. Because children's brains are still developing, especially in areas important for inhibition, children may be more susceptible than adults to tempting food cues. Objective: We examined potential developmental differences in children's and adults' responses to food cues to determine how these responses relate to weight status. Design: We included 27 children aged 10-12 y and 32 adults aged 32-52 y. Functional magnetic resonance imaging data were acquired during a food-viewing task in which unhealthy and healthy food pictures were presented. Results: Children had a stronger activation in the left precentral gyrus than did adults in response to unhealthy compared with healthy foods. In children, unhealthy foods elicited stronger activation in the right inferior temporal and middle occipital gyri, left precentral gyrus, bilateral opercular part of the inferior frontal gyrus, left hippocampus, and left middle frontal gyrus. Adults had stronger activation in the bilateral middle occipital gyrus and the right calcarine sulcus for unhealthy compared with healthy foods. Children with a higher body mass index (BMI) had lower activation in the bilateral dorsolateral prefrontal cortex while viewing unhealthy compared with healthy foods. In adults there was no correlation between BMI and neural response to unhealthy compared with healthy foods. Conclusions: Unhealthy foods might elicit more attention both in children and in adults. Children had stronger activation while viewing unhealthy compared with healthy foods in areas involved in reward, motivation, and memory. Furthermore, children activated a motivation and reward area located in the motor cortex more strongly than did adults in response to unhealthy foods. Finally, children with a higher BMI had less activation in inhibitory areas in response to unhealthy foods, which may mean they are more susceptible to tempting food cues.</p