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Effect of injected position of cultured cumulus cells as donor cells on reconstructed in vitro matured porcine oocytes

100%

Lin Y.-A., Tsai H.-B., Dybus A., Chen M.-C., Liao M.-H.

Animal Science Papers and Reports

2010

tom 28

nr 3

The effect was studied of injected position of cultured cumulus cells as donor cells for nuclear transfer on the process of in vitro matured reconstructed porcine oocytes. The cumulus-oocyte complexes (COCs) obtained from ovaries were cultured in NCSU-23 maturation medium. The cumulus cells were separated from matured COCs, and cultured in DMEM medium (10% fetal calf serum,FCS) for up to 4 passages. The cultured cumulus cells were injected into the enucleated oocytes to obtain the reconstructed oocytes. The chosen positions were the perivitelline space, ooplasm edge and ooplasm center. After nuclear transfer, the reconstructed oocytes were fused and activated simultaneously. Then, the activated oocytes were cultured in NCSU-23 embryo culture medium for 16 h to observe the activation, or 144 h to observe the embryo development. It was found that the injected position affected the activation rate, pronuclear formation rate and embryo development rate of the reconstructed oocytes significantly. The activation rate, pronuclear formation rate and embryo development rate of the ooplasm edge group were obviously greater than the perivitelline space and ooplasm center groups. Furthermore, the injected position at the ooplasm edge favoured the more advanced development. The ooplasm edge would be an optimum injected position.

Involvement of cyclooxygenase 2 and prostaglandin E2 in the effects of insulin on gastric emptying in male rats

72%

Huang W.-J., Hung C.-R., Chen M.-C., Wang J.-R., Doong M.-L., Wang C.-K., Hung C.-T., Wang P. S.

Journal of Physiology and Pharmacology

2009

tom 60

nr 3

109-118

Delayed gastric emptying in patients with both type 1 and type 2 diabetes mellitus (DM) occurs in approximately 50% of these patients. However, the role and the action mechanism of insulin on gastrointestinal (GI) motility are still unclear. The purpose of the present study was to investigate the involvement of cyclooxygenase-2 (COX-2) and prostaglandin E2 in the effects of insulin on gastric emptying in male rats. The normal and streptozotocin (STZ)-pretreated rats were injected intraperitoneally with or without insulin, atropine and specific muscarinic receptor antagonists before examination of measurement of gastric emptying, spontaneous contractile activity of smooth muscle strips, plasma cholecystokinin (CCK), and prostaglandin E2 (PGE2) analysis. Protein expression of COX-2 and insulin receptors (IRs) were analyzed by the technique of western blot. Acute different doses of insulin accelerated gastric emptying. Atropine interrupted the insulin effect on gastric emptying, and muscarnic M1/M3 receptor antagonists interrupted the insulin-reversed gastric emptying in normal and DM rats. Besides, we observed the expression of (IRs) in GI and found that IR was changed under the insulin and DM treatment, and was also different between STZ-pretreated rats and hyperglycemic rats. Expression of COX-2 in stomach was decreased in DM rats but restored by insulin. The COX inhibitor, indomethacin, decreased the gastric emptying which was induced or reversed by insulin in normal and DM rats, respectively. PGE2 production in stomach corresponded to the COX-2 expression. The contraction of GI smooth muscle stimulated by PGE2 was increased in insulin-pretreated normal and DM rats. We conclude that insulin changed the expression of IRs in stomach in DM rats. The delayed GI motility in diabetes was at least in part due to the COX-2 and PGE2 pathway which associated with decreasing COX-2 and diminishing PGE2 production in stomach. The attenuation of PGE2 production was employed for the index of the reduction of smooth muscle contraction in stomach in diabetes. Insulin stimulated the smooth muscle contraction through the IRs and COX-2 expression plus PGE2 production in rat stomach as well as reversed the delayed gastric emptying via the nervous actions of muscarinic M1 and M3 receptors in DM rats.

Ograniczanie wyników

1 Animal Science Papers and Reports

1 Journal of Physiology and Pharmacology

2 Chen M.-C.

1 Doong M.-L.

1 Dybus A.

1 Huang W.-J.

1 Hung C.-R.

1 Hung C.-T.

1 Liao M.-H.

1 Lin Y.-A.

1 Tsai H.-B.

1 Wang C.-K.

1 Wang J.-R.

1 Wang P. S.

1 2010

1 2009

Wyniki wyszukiwania

Effect of injected position of cultured cumulus cells as donor cells on reconstructed in vitro matured porcine oocytes

Involvement of cyclooxygenase 2 and prostaglandin E2 in the effects of insulin on gastric emptying in male rats