GSM8146664: B.gargarizans_PG, ScRNAseq; Bufo gargarizans; RNA-Seq - SRA

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SRX23945401: GSM8146664: B.gargarizans_PG, ScRNAseq; Bufo gargarizans; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 235.7M spots, 70.7G bases, 25.2Gb downloads

External Id: GSM8146664_r1

Submitted by: Chengdu University of TCM

Study: Mass spectrometry imaging and single-cell transcriptome reveal metabolic profiles of the postauricular glands of the Bufo gargarizans, and hypothesize Bufotoxin biosynthesis pathways

PRJNA1087441 • SRP495116 • All experiments • All runs

show Abstracthide Abstract

bufotoxin, which contain alkaloids, small peptides and bufadienolides (BDS), are neurotoxic and cardiotoxic, they are promising medical drugs, but the unclear specifics of bufotoxin in the postauricular glands (PG) and the biosynthetic routes of BDS are unknown. We used MALDI/MS to visualize 1872 components of PG, and established the expression profiles of 9316 cells by single-cell sequencing, which were classified into nine clusters by marker genes. We analyzed the expression differences of key genes for alkaloid, terpene backbone and steroid synthesis, identified optimal and silenced genes involved in key enzymes encoding bufotoxin biosynthesis, and hypothesized the full route of BDS synthesis via by analyzing the cholesterol metabolic pathway. BDS was enriched in EP, FB, and GG2 cells, and genes were expressed mainly in EP and FB cells. Combined with the bulk-RNA transcriptome we suggest that BDS synergistically synthesized by the liver and PG. metabolic mapping of toxin components in the PG of the toad, as demonstrated by our data. Overall design: The Bufo gargarizans' postauricular glands were separating into single-cell suspensions.

Sample: B.gargarizans_PG, ScRNAseq

SAMN40444448 • SRS20747285 • All experiments • All runs

Organism: Bufo gargarizans

Library:

Name: GSM8146664

Instrument: Illumina NovaSeq 6000

Strategy: RNA-Seq

Source: TRANSCRIPTOMIC SINGLE CELL

Selection: cDNA

Layout: PAIRED

Construction protocol: Adult toad postauricular glandular tissue was used, cut into approximately 1 mm pieces with sterilized scissors at room temperature, transferred into 15 ml centrifuge tubes, washed with PBS, and suspended in 3-4 ml of solution (TrypLE (1 mg /ml) containing collagenase I) and digested for 60 min until no tissue fragments were visible. To collect free cells, the tubes were centrifuged at 600 × g for 5 min at 4°C, the supernatant was removed and resuspended in 2 ml PBS. The dissociated cells were then passed through a 70 µm filter (Cat. No. 10) and then through a 40 µm filter (Cat. No. 10). After washing 2 times (centrifugation at 600 × g for 5 min at 4°C), they were resuspended in DPBS at a density of 1 × 105 cells/ml. Generation of single-cell RNA sequencing libraries and signature barcode matrices. library was performed according to the manufacter's instructing (Single Cell 3' V3.1, 10x genomices)

Runs: 1 run, 235.7M spots, 70.7G bases, 25.2Gb

Run	# of Spots	# of Bases	Size	Published
SRR28338831	235,705,388	70.7G	25.2Gb	2024-03-20

ID:: 32250690

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