Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 1. Immunofluorescence localization of EGFR in the hamster ovarian sections during the morning of Day 1, 0900 h (A); Day 3, 0900 h (C); Day 4, 0900 h (D); and Day 4, 1600 h (E). Green fluorescence represents EGFR while red fluorescence depicts the nucleus. Nuclear EGFR is indicated by arrowheads. Note no EGFR-specific signal is visible in the absence of the primary antibody (B). GC, Granulosa cells; Th, thecal cells; IC, interstitial cells; PF, primordial follicle; S1–S4, follicles with 1–4 layers of GC; S5–S6, follicles with 5–6 and 7–8 layers of GC, respectively; S8, small antral follicle; S10, large antral follicle; aGC, antral GC; mGC, mural GC; O, oocyte; SE, surface epithelium. Bars 5 10 mm except for A (5 20 mm) and E (5 5 mm).

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques:

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 2. EGFR immunofluorescence in the hamster ovary following hypophysectomy (A) and replacement of hCG (B), FSH (C), estrogen (D), and progesterone (E). Note marked increase in EGFR expression in mural GC following FSH administration (C). The presence of EGFR in the nuclei is evident following FSH or estradiol treatment (arrowheads). GC, Granulosa cells; Th, thecal cells; IC, interstitial cells; S0, primordial follicle; S1–S4, follicles with 1–4 layers of GC; S5–S6, follicles with 5–6 and 7–8 layers of GC, respectively; S10, large antral follicle; aGC, antral GC; mGC, mural GC; O, oocyte; aS6, atretic S6 follicle; SE, surface epithelium. Bars 5 10 mm except for C (S6) and D (5 5 mm).

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: Expressing

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 3. Northern blot of hamster ovarian EGFR mRNA.

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: Northern Blot

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 4. Expression patterns of EGFR mRNA in different ovarian cell com- partments during the estrous cycle. The density of silver grains was digi- tized following in situ hybridization. Bars with the same letters are not significantly (P , 0.05) different from each other.

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: Expressing, In Situ Hybridization

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 5. In situ hybridization detection of EGFR mRNA expression in the hamster ovary on Day 1, 0900 h (A); Day 3, 0900 h (C); Day 4, 0900 h (D); and Day 4, 1600 h (E). Note considerably higher signal intensity in granulosa cells of large antral follicles (S10) compared with smaller stages. No EGFR mRNA-specific signal could be detected in a Day 4, 1600 h ovarian section when sense EGFR cRNA was used as a probe (B). GC, Granulosa cells; Th, thecal cells; IC, interstitial cells; S5–S6, follicles with 5–6 and 7–8 layers of GC, respectively; S8–S10, small, medium, and large antral follicles, respectively; aGC, antral GC; mGC, mural GC; O, oocyte; AT, atretic follicle; SE, surface epithelium. Bars 5 30 mm.

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: In Situ Hybridization, Expressing

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 6. Expression patterns of EGFR mRNA in different ovarian cell com- partments following hypophysectomy and hormone replacement. The density of silver grains was digitized following in situ hybridization. Bars with the same letters are not significantly (P , 0.05) different from each other.

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: Expressing, In Situ Hybridization

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 7. In situ hybridization detection of EGFR mRNA expression in the hamster ovary following hypophysectomy (A) and FSH replacement (B–D). Note marked increase in EGFR mRNA expression in granulosa cells following FSH administration (B). Further, mural granulosa cells (mGC) expressed more EGFR mRNA compared with antral granulosa cells (aGC; C). Part D represents a bright-field micrograph of C. GC, Granulosa cells; Th, thecal cells; IC, interstitial cells; S5–S6, follicles with 5–6 and 7–8 layers of GC, respectively; S7, follicle with incipient antrum; S8–S10, small, medium, and large antral follicles, respectively; aGC, antral GC; mGC, mural GC; aS7, atretic S7 follicle. Bars 5 30 mm except for C and D (5 10 mm).

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: In Situ Hybridization, Expressing

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 8. In situ hybridization detection of EGFR mRNA expression in the hamster ovary following hCG (A), estrogen (B), progesterone (C), and estrogen 1 progesterone (D). GC, Granulosa cells; Th, thecal cells; IC, interstitial cells; S5–S6, follicles with 5–6 and 7–8 layers of GC, respectively; aGC, antral GC; mGC, mural GC; O, oocyte; AT, atretic follicle. Bars 5 30 mm.

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: In Situ Hybridization, Expressing

Journal: Biology of reproduction

Article Title: Spatiotemporal expression of epidermal growth factor receptor messenger RNA and protein in the hamster ovary: follicle stage-specific differential modulation by follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone.

doi: 10.1095/biolreprod.102.005470

Figure Lengend Snippet: FIG. 9. Semiquantitative RT-PCR evaluation of the steady-state levels of EGFR mRNA in the hamster ovary during the estrous cycle (A) and fol- lowing hypophysectomy and hormone replacement (B). DLU, Digital light unit. Bars with the same letters are not significantly (P , 0.05) different from each other.

Article Snippet: Anti-EGF-receptor (EGFR) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA); anti-phosphotyrosine antibody was purchased from Upstate Biotechnology (Upstate, NY); goat-anti-rabbit-IgGAlexa 488 (flurophor) was from Molecular Probes (Eugene, OR); nonimmune donkey serum was from Jackson Immunoresearch (West grove, PA); polymerase chain reaction (PCR) chemicals were from Roche Molecular Biochemicals (Indianapolis, IN), Amersham-Boehringer (Piscataway, NJ), and Promega (Madison, WI); PCR primers were synthesized by Genosys Biotechnologies (Woodlands, TX; [a-35S]CTP [specific activity, 800 Ci/mmol], was from ICN Radiochemicals (Costa Mesa, CA); and riboprobe synthesis kit was from Promega.

Techniques: Reverse Transcription Polymerase Chain Reaction