Unaltered Development of the Initial Follicular Waves and Normal Pubertal Onset in Female Rats after Neonatal Deletion of the Follicular Reserve

Abstract

In rats, the pool of primordial follicles is established within the first 3 d postnatally (dpn). Immediately after their differentiation, a subset of follicles begins to grow and constitutes the initial follicular waves. In this study we investigated the development of these early growing follicles after deletion of the primordial follicle pool induced by 1.5 Gy gamma-irradiation at 5 dpn. Within only 24 h, i.e. at 6 dpn, 99% of the primordial follicles disappeared, whereas most of the growing follicles remained unaffected. The study of these surviving follicles throughout the immature period has shown that their subsequent growth proceeded normally, as assessed by proliferating cell nuclear antigen immunostaining and follicular counts. No modification in the process of follicular atresia, studied by terminal deoxynucleotidyltransferase-mediated deoxy-UTP-fluorescein nick end labeling and Southern blot of DNA fragmentation analysis, was observed. Complementary analysis, by either in situ hybridization for inhibin subunits, P450 aromatase, and LH receptor mRNAs or plasma dosages of 17beta-estradiol and inhibin B, further showed that follicular maturation was unaltered. In line with these observations, pubertal onset was normal, regarding both age and ovulation rate. Nevertheless, as a consequence of the nonrenewal of the growing pool, the follicular complement was practically exhausted at puberty, and 90% of the females evidenced sterility by 4 months. Altogether, our results demonstrate that the deletion of the primordial follicle pool has induced no modification in the growth pattern of the early growing follicles that develop as their counterparts in control ovaries. Within the immature period, the initial follicular waves ensure the ovarian functionality and thus play a key role in the initiation of reproductive life.

The primordial follicle reserve is not renewed after chemical or γ-irradiation mediated depletion

“…Similar to previous reports for fetal or neonatal rats (Mazaud et al 2002, Guigon et al 2003, Mazaud Guittot et al 2006, we observed total elimination of primordial follicle numbers in mice after ionizing radiation. Interestingly, Johnson et al (2005) reported that bone marrow transplantation after exposure to g-irradiation was not able to rescue the ovarian failure (Johnson et al 2005).…”

Aromatase expression in the ovary: Hormonal and molecular regulation

“…Laser microdissection of preantral and antral follicles has recently confirmed these results [20]. Within the granulosa cell layer of antral follicles, aromatase expression is highest in the mural granulosa cells at the outer edge of the follicle when compared to granulosa cells closest to the antral cavity [14,21]. Moreover, aromatase is not expressed in cumulus granulosa cells [14,21].…”

“…Within the granulosa cell layer of antral follicles, aromatase expression is highest in the mural granulosa cells at the outer edge of the follicle when compared to granulosa cells closest to the antral cavity [14,21]. Moreover, aromatase is not expressed in cumulus granulosa cells [14,21]. The marked compartmentalization of aromatase expression within the granulosa layer is particularly striking in mature follicles from rats at proestrus (Fig.…”

“…Aromatase activity and mRNA can be found at days 5 and 8 after birth in the granulosa cell layer of growing follicles [10,14,15]. Aromatase expression progressively increases in both preantral and small antral follicles in infantile ovaries [Fig.…”

“…1a and reference 14]. In 21-dayold rats, aromatase is no longer expressed by growing follicles, but is confined to healthy large antral follicles [14]. Aromatase expression in neonatal ovaries requires normal levels of gonadotropins [16].…”