Prostaglandin synthesis by Day-6 rabbit blastocysts in vitro
in ReproductionSearch for other papers by M. J. K. Harper in
Current site
Google Scholar
PubMed
Search for other papers by M. A. Jones in
Current site
Google Scholar
PubMed
Search for other papers by C. J. Norris in
Current site
Google Scholar
PubMed
Search for other papers by D. S. Woodard in
Current site
Google Scholar
PubMed
Summary. Day-6 rabbit blastocysts were recovered from superovulated donor animals, washed in ice-cold Krebs–Ringer–bicarbonate (KRB) buffer, pooled and randomly allocated to polypropylene incubation tubes, usually 10 blastocysts in 1 ml KRB. The blastocysts were ruptured with a dissecting needle and incubated at 37°C for periods of 1–3 h with 10 μCi [3H]arachidonic acid/tube. A control tube without blastocysts was run in each experiment. At the end of the incubation, the samples were acidified, extracted with ethyl acetate, dried down and resuspended in h.p.l.c. column solvent. The radioactivity from the control tube eluting from the h.p.l.c., using a solvent system for prostaglandins (PGs), was subtracted from each experimental run in the same experiment. The remaining radioactivity constituted 0·14% of the original [3H]arachidonic acid added to each incubation tube. This was considered to have been the result of conversion of the radiolabelled arachidonic acid to prostanoids. In the absence of 10 mm-EDTA no conversion occurred, whereas in its presence peaks of radioactivity co-eluting with [3H]PGF-2α and [3H]PGE-2 were seen. A third peak that eluted was either 15-keto metabolites of these PGs or PGD-2. These 3 peaks were always significantly above background, and usually did not differ from each other. No differences in amount of conversion could be related to incubation time. Addition of indomethacin (100 μg/ml) or radioinert arachidonic acid (10 μg/ml) inhibited production of [3H]PG, even in the presence of EDTA. Removal of calcium from the incubation medium was per se without effect. Addition of atropine (0·15 mm) or carbachol (0·15 mm) in the presence or absence of EDTA did not change the pattern of conversion of [3H]arachidonic acid to [3H]PGs. These experiments demonstrate that rabbit blastocysts have the capacity for de-novo synthesis of PGs from exogenous substrate, when utilization of endogenous substrate is inhibited. The extent of conversion observed may not be a true reflection of the capacity for conversion of endogenous substrate.
Keywords: rabbit; blastocysts; prostaglandins
Reproduction is committed to supporting researchers in demonstrating the impact of their articles published in the journal.
The two types of article metrics we measure are (i) more traditional full-text views and pdf downloads, and (ii) Altmetric data, which shows the wider impact of articles in a range of non-traditional sources, such as social media.
More information is on the Reasons to publish page.
| Sept 2018 onwards | Past Year | Past 30 Days | |
|---|---|---|---|
| Full Text Views | 237 | 124 | 20 |
| PDF Downloads | 102 | 24 | 0 |
Online ISSN: 1741-7899
Print ISSN: 1470-1626
Strengthening biomedical communities
to advance science and health
CONTACT US
Bioscientifica Ltd | Starling House | 1600 Bristol Parkway North | Bristol BS34 8YU | UK
Bioscientifica Ltd | Registered in England no 3190519