|
| Status |
Public on Jan 25, 2024 |
| Title |
Germ cell from F3 sGC sample, biological rep 4 |
| Sample type |
RNA |
| |
|
| Source name |
F3 male germ cells, guinea pigs generated from F2 males that were generated from F1 males that were generated from F0 females exposed to sGC in late pregnancy
|
| Organism |
Cavia porcellus |
| Characteristics |
cell type: Germ cells from F3 male guinea pigs
Sex: male
treatment: SGC
|
| Treatment protocol |
Females (F0) were mated with males and once pregnant were treated with three courses of a clinically-relevant dose (1mg/kg) of betamethasone (Beta) or saline (Ctrl) on GD 40&41, 50&51, 60&61. Germ cells from F1 -F3 adult male offspring were collected using a density gradient followed by chemical and mechanical digestion F1_Ctrl(n=7), F1_sGC(n=6), F2(n=6/gp), and F3 (n=6/gp). Prefrontal cortext from adult female F2 (PND40) offspring was isolated and was stored in -80C prior to RNA extraction.
|
| Growth protocol |
Guinea pigs were singly housed on a 12-hour light-dark cycle. F0 females were mated with males to produce the F1 generation. F1 and F2 males were left undisturbed until adulthood and were mated with naiive females to generate F2 and F3 offspring, respectively. F1-F3 offspring were assigned to juvenile or breeding streams. Litters were weaned on PND 20 and pair-housed with an age-, sex- and treatment-matched partner. Food and water were available ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from germ cells and PFC using Trizol, according to the manufacturer’s recommendations, with some modifications. RNA was extracted from a total of 49 samples. Germ cells in F1-F3 adult male guinea pigs : F1_Ctrl(n=7), F1_sGC(n=6), F2(n=6/gp), and F3 (n=6/gp). PFC from F2 female guinea pigs (n=6/group).
|
| Label |
Biotin
|
| Label protocol |
1000ng of total RNA was used. Samples were labelled with FlashTag Biotin according to the manufacturer’s instructions
|
| |
|
| Hybridization protocol |
16 hours at 48C
|
| Scan protocol |
Affymetrix GeneChip Scanner 3000
|
| Description |
miRNA levels in F3 germ cells
|
| Data processing |
The data were analyzed with the Affymetrix Transcriptome Analysis Console 4.0.1(TAC) software using RMA+DMG. Differential analysis was conducted between control and sGC groups for each generation (F1-F3) in adult male germ cells, and for the F2 female PFC data, for a total of 4 analyses. All differential expression analyses were calculated using one-way ANOVA with Benjamini-Hochberg Step-UP FDR correction
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| |
|
| Submission date |
Dec 13, 2023 |
| Last update date |
Jan 25, 2024 |
| Contact name |
Christopher Casciaro |
| E-mail(s) |
chris.casciaro@mail.utoronto.ca, chris95@rogers.com
|
| Phone |
4169782025
|
| Organization name |
University of Toronto
|
| Department |
Department of Physiology
|
| Lab |
Dr. Stephen Matthews
|
| Street address |
1 King's College Circle
|
| City |
Toronto |
| State/province |
ON |
| ZIP/Postal code |
M5S 1A8 |
| Country |
Canada |
| |
|
| Platform ID |
GPL21572 |
| Series (1) |
| GSE250055 |
miRNA levels in F1-F3 male germ cells and female F2 PFC following prenatal exposure to sGC in guinea pigs |
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