# Food Restriction Induces Changes in Ovarian Folliculogenesis, Cell Proliferation, Apoptosis, and Production of Regulatory Peptides in Rabbits

## TL;DR

Reducing food intake in rabbits can enhance reproductive functions by improving follicle development and cell activity in the ovaries.

## Contribution

This study reveals how food restriction affects folliculogenesis and cell behavior in rabbit ovaries, offering new insights into reproductive physiology.

## Key findings

- Food restriction increased follicle and oocyte size and reduced apoptosis in granulosa cells.
- Restricted females showed higher levels of proteins related to cell proliferation and differentiation.
- Ovarian and uterine weights were lower in food-restricted rabbits compared to controls.

## Abstract

In prolific species such as rabbits, knowing how to improve their productive parameters through ovarian changes is of special importance. In this study, a 50% food restriction is proposed to induce changes in folliculogenesis, cell proliferation, and apoptosis. Females fed with 50% caloric restriction can improve fecundity by selection of the growing primordial ovarian follicles, better transformation of secondary to preovulatory follicles, and increased growth (cytoplasmic maturation) of oocytes. Caloric restriction can activate rabbit female reproduction by increasing proliferation and decreasing apoptosis in ovarian granulosa cells, changes in ovarian secretory activity, and changes in the number of peptides involved in cell differentiation, proliferation/division (cytoskeletal proteins), and adhesion.

The aim of this study is to examine the influence of food restriction on rabbit ovarian functions. A total of eight females were fed ad libitum (NF), while eight females were subjected to 50% food restriction (RF). One month later, all females were euthanized. Weights and lengths of ovaries and uterine horns were measured. Representative parts of the ovaries were subjected to histomorphometry analysis of folliculogenesis. Granulosa cells were isolated and cell viability, proliferation (accumulation of PCNA, cyclin B1, and BrdU-positive cells), apoptosis (accumulation of bax, caspase 3, and DNA fragmentation) were evaluated. Granulosa cells were subjected to proteomic analysis by using the nano HPLC-Chip-MS/MS method. Estradiol and progesterone release by ovarian and granulosa cells was assessed by ELISA. Ovarian and uterine horn weights were lower in RF than NF. The diameter of follicles and oocytes and the thickness of the theca and granulosa cells were higher in RF than NF. RF showed a lower percentage of cells containing bax and caspase 3, occurrence of DNA fragmented cells, and estradiol and progesterone. RF had higher incorporation of BrdU, a higher proportion of cells containing PCNA and cyclin B1, and a lower percentage of viable cells. RF produced more specific proteins than NF, including peptides involved in cell differentiation, proliferation/division, mitotic cell cycle, and GTP-ase activity. In conclusion, food restriction can activate reproduction by (1) selection of the growing primordial follicles, (2) better transformation of secondary to preovulatory follicles, (3) increasing growth of oocytes, (4) increasing proliferation and decreasing apoptosis in granulosa cells, (5) changes in ovarian secretory activity, and (6) changes in the number of peptides.