Journal: Poultry Science
Article Title: Heat stress enhances VLDL secretion in chicken ovarian follicles to potentiate its impact on follicular cell survival and maturation
doi: 10.1016/j.psj.2025.106137
Figure Lengend Snippet: Granulosa and theca cells respond to heat stress for VLDL-apoB secretion. Granulosa and theca cells were heat-stressed (HS, 42°C) for 3 hr (3H) and allowed recovery at 37°C for 5 (3H5R) or 13 hr (3H13R). Cells maintained at 37 °C served as a neutral control (NC). Medium collected at indicated time points was used for VLDL isolation. VLDL-apoB was determined by ELISA in isolated VLDL (panel A) and collected cells were used for apoB (panel B), MTTP-M (microsomal triglyceride transfer protein subunit M), and PDI (protein disulfide isomerase; MTTP-P subunit) expression by Western blotting (panel E), and for MTTP activity analysis (panel D). In metabolic labeling studies (panel C), cells were pre-treated with azidohomoalanine (AHA, a methionine analogue, 50 μM) overnight and then heat-stressed at 42°C for 3 hr and allowed recovery at 37°C. Medium were collected for VLDL isolation. The newly synthesized proteins identified by AHA incorporation in VLDL extracts were labeled with biotin azide and analyzed by regular Western blotting using streptavidin-HRP conjugate as a probe. Results of Western blot were normalized to β-actin and expressed as ratios relative to the control (NC 16H). *; significant effect by HS ( vs . NC at the same time point in panel A, or vs . NC 16H in panel B, D, and E), P < 0.05, n = 3. Means with different letters within the same thermal treatment (a, b, c) differ significantly among time points, P < 0.05, n = 3.
Article Snippet: VLDL-apoB concentrations were measured using a chicken-specific apoB ELISA kit (No. CSB-EL001918CH, CUSABIO, Houston, TX, USA).
Techniques: Control, Isolation, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Activity Assay, Labeling, Synthesized