Molecular Human Reproduction, Vol 3, 367-374, Copyright © 1997 by Oxford University Press
GD Palermo, OM Avrech, LT Colombero, H Wu, YM Wolny, RA Fissore and Z Rosenwaks
Among the possible mechanisms of oocyte activation after sperm penetration,
it appears most likely that a protein released by the spermatozoon elicits
a calcium elevation in the ooplasm. To further test this idea, cytosolic
factors obtained from human spermatozoa by two different methods,
freezing-thawing and sonication, were injected into mouse oocytes following
which intracellular calcium release was measured. Of a total of 42 mouse
oocytes, a pattern of calcium oscillations was observed in nine out of 16
oocytes injected with sonicated fraction, in all of eight oocytes with the
frozen-thawed fraction and in none of 18 control oocytes. Injection of the
frozen- thawed fraction also produced regular calcium oscillations in all
of five in-vitro matured human oocytes. To assess the putative factor's
ability to support fertilization, human oocytes that were not activated by
prior intracytoplasmic injection of spermatozoa (ICSI) and round spermatids
were reinjected with the frozen-thawed sperm fraction. Of 23 human oocytes
which remained unfertilized after ICSI, 19 became activated after injection
with sperm cytosolic factor; eight showed two pronuclei, three one
pronucleus and eight showed three or more pronuclei. Of 11 oocytes
unfertilized after prior round spermatid injection, two developed two
pronuclei, four developed one pronucleus and two had three or more
pronuclei. Cytogenetic analysis by fluorescence in-situ hybridization
confirmed the existence of a male pronucleus in eight out of nine such
zygotes displaying two or more pronuclei. Thus, human sperm extracts
activated mouse and human oocytes after injection, as judged by calcium
flux patterns in conjunction with male pronucleus formation.
JOURNAL ARTICLE
Human sperm cytosolic factor triggers Ca2+ oscillations and overcomes activation failure of mammalian oocytes
Center for Reproductive Medicine and Infertility, New York Hospital- Cornell Medical Center, New York 10021, USA.
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