Investigation of Oxtr-expressing Neurons Projecting to Nucleus Accumbens Essay
https://doi.org/10.1016/j.neuroscience.2020.08.023
Investigation of Oxtr-expressing Neurons Projecting to Nucleus Accumbens using Oxtr-ires-Cre Knock-in prairie Voles (Microtus ochrogaster)
questions
1) How did the authors identify oxytocin-expressing neurons that project into the nucleus accumbens (NAcc)? ( mcq) options are ?
a)They generated transgenic voles expressing a fluorescent protein under the control of a NAcc-specific promoter and co-stained for oxytocin .
b)They made transgenic voles expressing Cre recombinase under the control of the oxytocin promoter and introduced a CRE-dependent luminescent gene to neurons by infusing a virus into the nucleus accumbens
c)They made transgenic voles expressing Cre recombinase under the control of the oxytocin promoter and introduced a CRE-dependent luminescent gene to neurons by infecting the entire vole with a virus
d)They made transgenic voles expressing Green Fluorescent Protein under the control of the adeno-associated virus promoter and then infused adeno-associated virus into the nucleus accumbens
2) What transgene did the researchers introduce to voles?
What gene’s promoter controls it?
What system/technique was essential in generating the voles?
3) What technique did the authors use to show that Cre and Otxr are colocalized in the brain?
4) Did the researchers use homozygous Cre/Cre voles or the heterozygous +/Cre in the experiments? What was the reason for choosing one and not the other?
5) What is “AAVrg-hSyn-DIO-EGFP”? It’s used in these experiments to trace neurons; how does it get into them?
6) In what regions of the brain (aside from the nucleus accumbens) do the researchers find cell bodies of oxytocin receptor-expressing neurons that extend into the nucleus accumbens?
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7) Pick one region from the last question and describe what its function is. ?
8) What are the functions of the prelimbic cortex?
What do you think is the significance of oxytoxin receptor-expressing neurons extending from the prelimbic cortex to the nucleus accumbens?
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9) What are the functions of the olfactory bulb?
What do you think is the significance of oxytoxin receptor-expressing neurons extending from the olfactory bulb to the nucleus accumbens? Investigation of Oxtr-expressing Neurons Projecting to Nucleus Accumbens Essay.
Social bonds such as parent-infant attachment or pair bonds can be critical for mental and physical well-being. The monogamous prairie vole (Microtus ochrogaster) has proven useful for examining the neural substrates regulating social behaviors, including social bonding. Oxytocin (OXT) and oxytocin receptor (OXTR) play critical roles in alloparental care, pair bonding and consoling behavior in prairie voles. While OXTR in a few regions, such as the nucleus accumbnes (NAcc), prefrontal cortex (PFC) and anterior cingulate cortex (ACC), have been implicated in regulating these behaviors, the extent to which other OXT sensitive areas modulate social behaviors has not been investigated. The NAcc is a central hub for modulating OXTR dependent social behaviors. To identify neurons expressing Oxtr in prairie vole brain, we generated gene knock-in voles expressing Cre recombinase in tandem with Oxtr (Oxtr-ires-Cre) using CRISPR/Cas9 genome editing. We confirmed Oxtr and Cre mRNA co-localization in NAcc, validating this model. Next, we identified putative Oxtr-expressing neurons projecting to NAcc by infusing retrograde CRE-dependent EGFP AAV into NAcc and visualizing fluorescence. We found enhanced green fluorescent protein (EGFP) positive neurons in anterior olfactory nucleus, PFC, ACC, insular cortex (IC), paraventricular thalamus (PVT), basolateral amygdala (BLA), and posteromedial and posterolateral cortical amygdaloid area (PMCo, PLCo). The ACC to NAcc OXTR projection may represent a species-specific circuit since Oxtr-expressing neurons in the ACC of mice were reported not to project to the NAcc. This is the first delineation of Oxtr-expressing neural circuits in the prairie vole, and demonstrates the utility of this novel genetically modified organism for characterizing OXTR circuits involved in social behaviors. Investigation of Oxtr-expressing Neurons Projecting to Nucleus Accumbens Essay.
