Sample 17 HTO
Rattus norvegicus
Protocols: For all experiments that involve RNA sequencing, brains were extracted after sacrifice and the hindbrain portion containing the DVC was dissected out before being fresh frozen on powdered dry ice for intermediate storage in 1.5ml Eppendorf tubes at -80°C. These dissections were later homogenized and incubated in EZ lysis buffer, and subsequently strained through a 40µm filter and spun down DIO mice were allocated into groups based on BW. For the acute study, mice were dosed with a single subcutaneous injection of cagrilintide (10 nmol kg-1) or vehicle 0-1 hour after the onset of the light cycle. The mice were euthanized by decapitation under isoflurane anaesthesia 3-4 h after dosing. For the subchronic study, mice were dosed subcutaneously once a day with cagrilintide (10 nmol kg-1) or vehicle 3-4 h after onset of light for 7 days. One group of mice was weight-matched to mice dosed with cagrilintide by daily food restriction  Frozen tissue was thawed on ice and homogenized in Nuclei EZ Lysis Buffer using a glass tissue douncer. The homogenate was incubated on ice, filtered through a 40 µm cell strainer, and centrifuged. The pellet was resuspended in Nuclei EZ Lysis Buffer and OptiPrep, layered onto 49% OptiPrep, and centrifuged for density purification. Isolated nuclei were collected, washed in nuclei buffer containing PBS, 1% BSA, 2 mM MgCl2, and RNase inhibitor, centrifuged again, and finally resuspended in nuclei buffer. This procedure generated single-nucleus suspensions for downstream hashing and snRNA-seq library preparation. Single-nucleus suspensions were hashed by incubation with TotalSeq-A anti-nuclear hashtag oligonucleotide-conjugated antibodies on ice, followed by washing and resuspension in nuclei buffer containing DAPI. DAPI-positive nuclei were sorted by flow cytometry and collected into RT Buffer B. The sorted nuclei suspension was adjusted to the required input volume and processed using the Chromium Next GEM Single Cell 3′ Kit v3.1 (10x Genomics) according to the manufacturer's instructions. A multiplexing hashtag library was generated following the BioLegend TotalSeq-A Cell Hashing protocol for compatibility with the 10x Genomics Single Cell 3′ Reagent Kit v3.1.
ENA first public
2026-06-09
INSDC center name
Novo Nordisk Foundation Center for Basic Metabolic Research
INSDC status
public
Submitter Id
E-MTAB-16929:Sample 17 HTO
age
25
broker name
ArrayExpress
cell type
neuron and glia
collection date
not collected
common name
Norway rat
developmental stage
adult
genotype
wild type genotype
geographic location (country and/or sea)
not collected
hashtag index id(s)
D706, D707, D708, D709, D710, D702
individual
pooled
isolate
not applicable
organism part
dorsal vagal complex
protocol
HTO
run id
20220621_0285
sample name
E-MTAB-16929:Sample 17 HTO
scientific_name
Rattus norvegicus
sex
male
strain
Sprague-Dawley