Document Detail


Multiple classes of prostaglandin F2 alpha binding sites in subpopulations of ovine luteal cells.
MedLine Citation:
PMID:  2590710     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A cryostorage procedure was developed to provide ovine luteal cells throughout the period of seasonal anestrus. Corpora lutea obtained from midluteal phase, superovulated ewes were dispersed enzymatically. Some dispersed cells were fractionated into subpopulations by elutriation. Dimethylsulfoxide (7.5% final concentration) in Hanks' buffered saline was added to cells at 4 degrees C, and dispersed cell preparations were frozen in a programmable cell freezer and stored at -196 degrees C. After recovery from cryopreservation, cell viability and prostaglandin F2 alpha (PGF2 alpha) binding characteristics of thawed cells were not different from those of corresponding fresh cells. Additionally, thawed cells retained the capacity to attach to culture dishes and retained responsiveness of progesterone secretion to prostaglandin E2 (PGE2) and ovine luteinizing hormone (LH), although rates of progesterone secretion were attenuated in thawed compared with fresh cells. The cryopreservation procedure will prove useful to relieve constraints in utilization of ovine luteal cells arising from reproductive seasonality in sheep. Cells retrieved from cryostorage were evaluated by studying PGF2 alpha binding characteristics. From saturation analyses (increasing amounts of radiolabeled PGF2 alpha) of PGF2 alpha binding to unfractionated cells, we detected a single class of high affinity binding sites (Kd = 17.4 +/- 2.3 nM) in addition to the nonspecific binding component. Using displacement analyses (constant radiolabeled PGF2 alpha and increasing amounts of unlabeled PGF2 alpha) and unfractionated cells, we detected additional binding sites of lower affinity (Kd = 409 +/- 166 nM) as well as the nonspecific binding component. Small luteal cells obtained by elutriation, which were essentially devoid of large cell contamination, had only low affinity binding sites.(ABSTRACT TRUNCATED AT 250 WORDS)
Authors:
A K Balapure; I C Caicedo; K Kawada; D S Watt; C E Rexroad; T A Fitz
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biology of reproduction     Volume:  41     ISSN:  0006-3363     ISO Abbreviation:  Biol. Reprod.     Publication Date:  1989 Sep 
Date Detail:
Created Date:  1990-01-25     Completed Date:  1990-01-25     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0207224     Medline TA:  Biol Reprod     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  385-92     Citation Subset:  IM    
Affiliation:
Department of Obstetrics and Gynecology, Uniformed Services, University of the Health Sciences, Bethesda, Maryland 20814.
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MeSH Terms
Descriptor/Qualifier:
Animals
Binding Sites
Cell Fractionation
Cell Survival
Corpus Luteum / metabolism*
Cryopreservation
Dinoprost / metabolism*
Female
Luteal Cells / cytology,  metabolism*
Male
Progesterone / secretion
Sheep
Grant Support
ID/Acronym/Agency:
HD20780/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
551-11-1/Dinoprost; 57-83-0/Progesterone

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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