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

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: Effects of Lomitapide and Mipomersen on VLDL secretion of granulosa and theca cells under heat stress. Granulosa and theca cells were treated with Lomitapide (Lom) and Mipomersen (Mip) (MTTP and ApoB inhibitor, respectively, 2.5 μM for each) for 2 hr. After washout, cells were heat-stressed (HS, 42°C) for 3 hr (3H) and allowed recovery at 37°C for 13 hr (3H13R). Cells maintained at 37°C served as a neutral control (16H NC). Medium were collected for VLDL isolation and then used for VLDL-apoB determination through the ELISA method (panel A) and cells were used for MTTP activity analysis (panel C). In metabolic labeling studies (panel B), cells pre-treated with azidohomoalanine (AHA, 50 μM) overnight and then with Lom and Mip for 2 hr were subjected to HS treatment at 42°C for 3 hr and allowed recovery at 37°C. Medium were collected for VLDL isolation. AHA incorporation in VLDL protein extracts was labeled with biotin azide and analyzed by the regular Western blot method. *; significant effect by HS ( vs . NC 16H within the same pharmacological treatment), +; significant effect by Lom+Mip ( vs . vehicle within the same thermal treatment at the same time), 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, Activity Assay, Labeling, Western Blot

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: Effects of Lomitapide and Mipomersen on sex steroid secretion of granulosa and theca cells under heat stress. Granulosa and theca cells were treated with Lomitapide (Lom) and Mipomersen (Mip) (2.5 μM for each) for 2 hr. After washout, cells were heat-stressed (HS, 42°C) for 3 (3H, panel A) or 8 hr (8H, panel B) and allowed recovery for 13 (3H13R) or 8 hr (8H8R), respectively. Medium collected at indicated time points were used for progesterone (P4) and estradiol (E2) determination through ELISA kits. *; significant effect by HS ( vs . NC within the same pharmacological treatment at the same time), #; significant effect by time ( vs . 3H within the same thermal and pharmacological treatment), +; significant effect by Lom+Mip ( vs. vehicle within the same thermal treatment at the same time), 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: Enzyme-linked Immunosorbent Assay