Apoptosis Modulation on Porcine Granulosa Cells by a GnRH Analog

FIORITO, C.; CAROU MC.; REVILLA M.; LOMBARDO DM.

INTERNATIONAL JOURNAL OF MORPHOLOGY

Universidad Nacional de La Frontera

Lugar: Ciudad de Temuco. Chile; Año: 2008 vol. 26 p. 726 - 727

0717-9367

<!-- /* Font Definitions */ @font-face {font-family:"Cambria Math"; panose-1:2 4 5 3 5 4 6 3 2 4; mso-font-charset:0; mso-generic-font-family:roman; mso-font-pitch:variable; mso-font-signature:-1610611985 1107304683 0 0 159 0;} /* Style Definitions */ p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-unhide:no; mso-style-qformat:yes; mso-style-parent:""; margin:0cm; margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman","serif"; mso-fareast-font-family:"Times New Roman";} a:link, span.MsoHyperlink {mso-style-unhide:no; color:blue; text-decoration:underline; text-underline:single;} a:visited, span.MsoHyperlinkFollowed {mso-style-noshow:yes; mso-style-priority:99; color:purple; mso-themecolor:followedhyperlink; text-decoration:underline; text-underline:single;} .MsoChpDefault {mso-style-type:export-only; mso-default-props:yes; font-size:10.0pt; mso-ansi-font-size:10.0pt; mso-bidi-font-size:10.0pt;} @page WordSection1 {size:612.0pt 792.0pt; margin:70.85pt 3.0cm 70.85pt 3.0cm; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.WordSection1 {page:WordSection1;} --> APOPTOSIS MODULATION ON PORCINE GRANULOSA CELLS BY A GnRH ANALOG. Fiorito, C., Carou MC., Revilla M.; Lombardo DM. Histología y Embriología. Facultad de Cs. Veterinarias, UBA. E-Mail: dlombard@fvet.uba.ar   Granulosa cells (GC) contribute on ovocit maturation and most of the growth factors that it needs act through them. This makes that GC viability and integrity to be an important factor throughout follicle genesis. It’s known that GC death during follicular atresia and luteolisys is apoptotic, although, the regulation process is not yet well understood. Gonadotropin releasing hormone receptor (GnRHr) was found on GC and luteal cells of various species. There was observed a decrease on GCs proliferate activity on porcine immature follicles and an increase on apoptosis levels independently on follicle genesis stage.  Our objective is to determine if a GnRHa (Leuprolide Acetate) is an apoptotic modulator on GC from adult porcine’s ovaries and also precise the best experimental conditions to observe LA modulation on this model.  This study was carried on a primary GC porcine culture (CPGP) model obtained from punction of 5-8 nm follicles. CPGPs were grown in DMEM + F12 supplemented with SFB 5% and antibiotics.  We assay three LA concentrations (1, 10 and 100 nM) and two incubation times (12 and 24 Hs). Three experiments by triplicate were purchased and were measured for apoptosis with DAPI, TUNEL and by a citometric analysis (FACS) using Anexina V and PI label techniques.  Results were statistical analyzed with a two way ANOVA and Bonferroni comparison of means.  TUNEL and DAPI techniques show that 100 nM LA for 24 Hs treatment an 27 % augment of apoptosis over control (p = 0.058). This effect was corroborated by FACS with Anexina V and PI label, being, the differences, significant for LA 1 nM for 24 Hs treatment, with an apoptosis increment of 74 % over control (p = 0.05). FACS analysis shows a characteristic apoptotic temporal dynamic on CPGPs exposed to LA for this model on this experimental conditions.