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Detection and Isolation of Apoptotic Bodies to High Purity

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Subpopulations of physiological and ovarian follicular fluid peptide induced apoptotic cells

M S Rajadhyaksha1, T D Nandedkar

  • 1Department of Life Sciences, Sophia College, Mumbai, India.

Indian Journal of Experimental Biology
|April 28, 2000
PubMed

Insights

Ovarian follicular fluid peptide (OFFP) induces apoptosis in mouse ovarian granulosa cells. OFFP affects distinct cell subpopulations compared to natural atresia, with reduced FSH binding in treated cells.

Area of Science:

  • Reproductive Biology
  • Cell Biology
  • Endocrinology

Background:

  • Ovarian follicular fluid peptide (OFFP) from sheep ovaries is known to induce apoptosis in mouse ovarian granulosa cells.
  • Understanding the specific granulosa cell subpopulations affected by OFFP is crucial for elucidating its mechanism of action.

Purpose of the Study:

  • To investigate the heterogeneity of ovarian granulosa cells in response to OFFP.
  • To compare the granulosa cell subpopulations involved in OFFP-induced apoptosis versus physiological atresia.
  • To assess the impact of OFFP on FSH binding to granulosa cells.

Main Methods:

  • Granulosa cells were isolated from immature mice treated with PMSG (control, atretic) or PMSG followed by OFFP.
  • Cell subpopulations were separated using discontinuous Percoll gradient centrifugation.
  • Apoptotic cells were quantified using acridine orange staining.
  • Follicle-stimulating hormone (FSH) binding was measured by radio receptor assay.

Main Results:

  • Distinct granulosa cell subpopulations responded to OFFP-induced apoptosis compared to those in physiological atresia.
  • OFFP treatment led to significantly lower FSH binding in granulosa cells compared to control and atretic groups.
  • Differences in cell densities suggest distinct subpopulations or apoptotic phases are involved.

Conclusions:

  • OFFP induces apoptosis in specific granulosa cell subpopulations, differing from those in natural follicular atresia.
  • OFFP treatment may alter granulosa cell responsiveness to FSH, indicated by reduced binding.
  • Further research is needed to understand the implications of these distinct subpopulations and altered FSH binding for ovarian function.

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