Mus musculus strain:ChAT-Cre/ZsGreen
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA901862 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR22307344.fastq.gz | Fastqsanger.gz | |||
| SRR22307345.fastq.gz | Fastqsanger.gz | |||
| SRR22307346.fastq.gz | Fastqsanger.gz | |||
| SRR22307347.fastq.gz | Fastqsanger.gz | |||
| SRR22307348.fastq.gz | Fastqsanger.gz |
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2021-10-31 | E-MTAB-8760 | biostudies-arrayexpress
Project description:As part of the NIH SPARC program efforts to study the intrinsic cardiac nervous system, we developed the first comprehensive molecular phenotyping data on the right atrial ganglionated plexus (RAGP) within the intrinsic cardiac nervous system of the pig. We collected hundreds of single neurons from the RAGP and assayed these for expression of a wide range of genes relevant to neuronal functions. The sinoatrial node (SAN) of 4 pigs were injected with FastBlue, a retrograde tracer, labeling neurons projecting to the SAN. The RAGP of those pigs were then collected and sectioned. Both SAN-projecting and non-SAN-projecting single neurons in RAGP were collected through laser capture microdissection (LCM), allowing for spatial mapping of all collected samples. Over 400 single neurons were collected (n=4 animals) and each neuron was assayed for over 300 genes using high throughput microfluidic qPCR using BioMark. Neuronal phenotypes were distinguished by multivariate analysis showing patterns of network activity between multiple genes. Interestingly, these phenotypes show no spatial preferences within the RAGP. Our results revealed extensive combinatorial expression of neurotransmitters across the RAGP neuronal phenotypes. Additionally, there was a large overlap in expression profiles of SAN-projecting and non-SAN-projecting neurons, without any single gene or module acting as a distinguishing marker between the connectionally labeled groups. Previous studies have focused on cholinergic and catecholaminergic processes, showing evidence for extensive protein expression of cholinergic markers in RAGP. Our findings are in stark contrast to these results and demonstrate high gene expression correlation between cholinergic and catecholaminergic markers such as tyrosine hydroxylase (Th) and choline acetyltransferase (ChAT) in single neurons. This finding suggests that cells are poised for a rapid shift in neurotransmitter signaling as well as the possibility for post transcriptional regulation of neuron phenotype plasticity. Our present findings significantly expand the list of neuromodulatory peptides and their receptors known to be expressed in RAGP. For example, neuropeptide Y and somatostatin showed distinct co-expression patterns with their respective receptors with significant non-overlapping expression of transmitters and cognate receptors across single neurons. Taken together, our results reveal a complex organization of neuronal networks within RAGP-SAN complex that is not entirely governed by connectivity or spatial location, with combinatorial patterns of local paracrine networks with potential influence on cardiac function.
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Project description:The cholinergic system is a critical regulator of Pavlovian fear learning and extinction. As such, we have begun investigating the cholinergic system's involvement in individual differences in cued fear extinction using a transgenic ChAT::Cre rat model. The current study extends behavioral phenotyping of a transgenic ChAT::Cre rat line by examining both freezing behavior and ultrasonic vocalizations (USVs) during a Pavlovian cued fear learning and extinction paradigm. Freezing, 22 kHz USVs, and 50 kHz USVs were compared between male and female transgenic ChAT::Cre+ rats and their wildtype (Cre-) littermates during fear learning, contextual and cue-conditioned fear recall, cued fear extinction, and generalization to a novel tone. During contextual and cued fear recall ChAT::Cre+ rats froze slightly more than their Cre- littermates, and displayed significant sex differences in contextual and cue-conditioned freezing, 22 kHz USVs, and 50 kHz USVs. Females showed more freezing than males in fear recall trials, but fewer 22 kHz distress calls during fear learning and recall. Females also produced more 50 kHz USVs during exposure to the testing chambers prior to tone (or shock) presentation compared with males, but this effect was blunted in ChAT::Cre+ females. Corroborating previous studies, ChAT::Cre+ transgenic rats overexpressed vesicular acetylcholine transporter immunolabeling in basal forebrain, striatum, basolateral amygdala, and hippocampus, but had similar levels of acetylcholinesterase and numbers of ChAT+ neurons as Cre- rats. This study suggests that variance in behavior between ChAT::Cre+ and wildtype rats is sex dependent and advances theories that distinct neural circuits and processes regulate sexually divergent fear responses.
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