02631nam a2200313 a 450000100080000000500110000800800410001910000190006024501050007926000530018450001210023752016800035865000100203865000140204865000160206265000120207865000200209065000140211065300190212465300160214365300160215965300240217565300230219970000180222270000170224070000180225770000220227570000200229715149362021-09-08       1997    bl uuuu     u00u1 u    #d1 aKESKINTEPE, L.  aCaprine blastocyst development after intracytoplasmic sperm injection (ICSI).h[electronic resource]  aTheriogenology, v. 47, n.1 , p. 249, 1997.c1997  aProceedings Annual Conference of the International Embryo Transfer Society, Nice Acropolis, Nice, France, Jan. 1997.  aExperiments were undertaken to develop ICSI to produce caprine embryos outside the normal breeding season. Oocytes with 2-4 layers of cumulus cells from 2-6 mm follicles were washed twice with Tyrodes + 0.4 mg PV A and 0.4 mg sodium pyruvate per ml and incubated in 1 ml of Hepes-TCM 199 + 10 J.Lg each oFSH and bLH (NHPP, NIDDK, NICHD, USDA) + 20% FBS (MM) in a thermos (38.5°C) for 4.5 h. Then, oocytes were transferred into 75 ul of freshly prepared MM under paraffin oil and 5% O2, 5% CO2, and 90% N2. Approximately 26 h after recovery oocytes were denuded with hyaluronidase (100 lU/mi) and pipetting and held at 38.5°C for 90 min. Sperm frozen in egg yolk extender were thawed in a 37°C water bath for 15 sec, and motile fractions were selected by swim-up, then incubated 90 min in TALP with 10 J.tg heparin/ml. Each oocyte was positioned with its first polar body at 6 or 12 0' clock by a holding pipette. Sperm medium (1 J.LI) was added to 10 J.tl medium containing 10% PVP. A sperm cell was aspirated into a pipette, and then injected head first into the cytoplasm of an oocyte in H- TCM 199 + 20% FBS (HM) at 37°C. Injected oocytes were transferred to HM and after 90 min, cultured in 50 J.tl of c- SOF+NEA (Biol. Reprod.52: 141 0-1417). Data analysis was by ANOV A and Bonferroni t-test. Tabulated results show that caprine blastocysts ca~ be derived out of season after use of frozen- thawed sem en and injection of sperm cells with broken tails into ooplasm. Controls included comparable numbers of oocytes activated with Ca ionophopre (5 mM for 1 min), injected without sperm, and injected without sperm and activated by Ca ionophore, none of which cleaved.  aGoats  aInjection  aSpermatozoa  aCaprino  aEspermatozóide  aInjeção  aAnimal embryos  aBlastocisto  aBlastocysts  aIntracitoplasmatico  aIntracytoplasmatic1 aMORTON, P. C.1 aSMITH, S. E.1 aTUCKER, M. J.1 aSIMPLÍCIO, A. A.1 aBRACKETT, B. G.