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Regular Article |
a Dipartimento di Morfofisiologia Veterinaria e Produzioni Animali, Fisiologia Veterinaria, Università di Bologna, 40064 Bologna, Italy
b Dipartimento di Strutture, Funzioni e Patologie Animali e Biotecnologie, Fisiologia Veterinaria, Università di Teramo, 64020 Teramo, Italy
c Dipartimento di Strutture, Funzioni e Patologie Animali e Biotecnologie, Anatomia Normale Veterinaria, Università di Teramo, 64020 Teramo, Italy
ABSTRACT
The authors evaluated the relationship between vascular endothelial growth factor (VEGF) production, blood vessel extension, and steroidogenesis in small (<4 mm), medium (4–5 mm), and large (>5 mm) follicles isolated from gilts treated with eCG. VEGF and estradiol levels were measured in follicular fluid by an enzyme immunoassay and radioimmunoassay, respectively, and then each follicle wall was used to evaluate VEGF mRNA content and for the immunohistochemical analysis of blood vessels. VEGF production was low in small follicles (<3 ng/ml), high in large follicles (>10 ng/ml), and markedly differentiated in medium follicles; 44% exhibited values up to 15 ng/ml, whereas the levels never exceeded 3 ng/ml in the remaining aliquot. Medium follicles were then used as a model to investigate angiogenesis. Reverse transcription-polymerase chain reaction for VEGF mRNA demonstrated that granulosa cells represent the main component involved in the production of VEGF. The follicle wall, which presents two distinct concentric vessel networks, showed a vascular area (positive stained area/percent of field area) that was significantly wider in high VEGF follicles than in low VEGF follicles (2.54% ± 0.58% vs. 1.29% ± 0.58%, respectively). Medium follicles with high VEGF levels and extensive vascularization accumulated high estradiol levels (150–300 ng/ml), whereas follicles with low VEGF levels had basal estradiol levels that never exceeded 30 ng/ml. Early atretic medium-size follicles had undetectable levels of VEGF and estradiol paralleled by a marked reduction in blood vessel. The data presented propose an improved model for follicle dynamics in which the production of VEGF, stimulated by gonadotropin, creates the vascular conditions required for follicle growth and activity.
1 This work was supported by cofinanziamento MURST Es fin. 1999.
2 Correspondence: Mauro Mattioli, Dipartimento di Morfofisiologia Veterinaria e Produzioni Animali, Università di Bologna, Via Tolara di Sopra, 50, Ozzano Emilia, 40064 Bologna, Italy. FAX: 0861 558819; mattioli{at}vet.unibo.it
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