Effect of ovine luteinizing hormone (oLH) charge isoforms on VEGF and cAMP production.
Although an increase in VEGF expression and synthesis in association with LH has been established; it is unknown if all LH isoforms act similarly. This study evaluated the production of cAMP and VEGF among LH isoforms in two in vitro bioassays. The LH was obtained from hypophyses and the group of isoforms was isolated by chromatofocusing. cAMP production was assessed using the in vitro bioassay of HEK-293 cells and VEGF production was evaluated in granulosa cells. Immunological activity was measured with a homologous RIA. Immunoactivity and bioactivity for each isoform were compared against a standard, by estimating the IC50 and the EC50. The basic isoforms were more immunoactive than the standard. The neutral and the moderately acidic had an immunological activity similar to the standard. The acidic isoform was the least immunoreactive. cAMP production at the EC50 dose was similar among the basic isoforms, the moderately acidic and the standard; for the neutral and the acidic, the EC50 dose was higher. It was observed that compared with the control, VEGF production at the lowest LH dose was no different in the standard and each isoform. In the intermediate dose, a positive response was caused in the standard and the neutral and basic isoforms. Although the acidic isoform showed a dose-dependent response, it was not significant relative to the control. In conclusion, the basic isoform generated the greatest cAMP and VEGF production, similar to the reference standard, and the acidic the smallest.
Departamento de Reproducción, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico.
Arnulfo
A
Montero-Pardo
Departamento de Biología Celular y Fisiología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico.
Daniel
D
Diaz
Unidad de Investigación Médica en Medicina Reproductiva, UMAE Hospital de Gineco Obstetricia No. 4 Luis Castelazo Ayala, IMSS, México D.F., Mexico.
Aleida
A
Olivares
Departamento de Reproducción, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico. Electronic address: ever@unam.mx.
Everardo
E
González-Padilla
Departamento de Reproducción, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico.
Clara
C
Murcia
Departamento de Biología Celular y Fisiología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico.
Margarita
M
Gómez-Chavarín
Departamento de Biología Celular y Fisiología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico.
Gabriel
G
Gutiérrez-Ospina
Departamento de Reproducción, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico; Departamento de Biología Celular y Fisiología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Mexico. Electronic address: pererag@unam.mx.
Gerardo
G
Perera-Marín
D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D007986:Luteinizing Hormone / Q000737:chemistry / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects; D010903:Pituitary Gland, Anterior; D020033:Protein Isoforms; D012756:Sheep / Q000502:physiology*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D020033:Protein Isoforms; D042461:Vascular Endothelial Growth Factor A; D007986:Luteinizing Hormone; D000242:Cyclic AMP
Granulosa; In vitro cAMP; Isoforms; Luteinizing hormone; Ovine; VEGF