21 research outputs found
Acrosome components after intracytoplasmic sperm injection: the decondensation frontier
http://www.sciencedirect.com/science/article/B6T6K-43CBFGC-14/1/c122d3f8e7188ef9ec4a133a8068995
Central spindle self-organization and cytokinesis in artificially activated sea urchin eggs
Author Posting. © Marine Biological Laboratory, 2016. This article is posted here by permission of Marine Biological Laboratory for personal use, not for redistribution. The definitive version was published in Biological Bulletin 230, no.2 (2016): 85-95.The ability of microtubules of the mitotic apparatus
to control the positioning and initiation of the cleavage
furrow during cytokinesis was first established from
studies on early echinoderm embryos. However, the identity
of the microtubule population that imparts cytokinetic signaling
is unclear. The two main––and not necessarily mutually
exclusive–– candidates are the central spindle and the
astral rays. In the present study, we examined cytokinesis in
ammonia-activated sea urchin eggs, which lack paternally
derived centrosomes and undergo mitosis mediated by unusual
anastral, bipolar mini-spindles. Live cell imaging and
immunolabeling for microtubules and the centralspindlin
constituent and kinesin-related protein, MKLP1, demonstrated
that furrowing in ammonia-activated eggs was associated
with aligned arrays of centralspindlin-linked, opposed
bundles of antiparallel microtubules. These autonomous, zipper-
like arrays were not associated with a mitotic apparatus,
but did possess characteristics similar to the central spindle
region of control, fertilized embryos. Our results highlight the
self-organizing nature of the central spindle region and its
ability to induce cytokinesis-like furrowing, even in the absence
of a complete mitotic apparatus.This research was
supported by student/faculty summer research grants from
the Dickinson College Research and Development Committee
to JHH; Laura and Arthur Colwin Summer Research
Fellowships from the MBL to JHH and CBS; a National
Science Foundation Major Research Instrumentation grant
to JHH (MRI-0320606); and a NSF collaborative research
grant to JHH (MCB-1412688) and to CBS (MCB-
1412734)
Sperm Chromatin-Induced Ectopic Polar Body Extrusion in Mouse Eggs after ICSI and Delayed Egg Activation
Meiotic chromosomes in an oocyte are not only a maternal genome carrier but also provide a positional signal to induce cortical polarization and define asymmetric meiotic division of the oocyte, resulting in polar body extrusion and haploidization of the maternal genome. The meiotic chromosomes play dual function in determination of meiosis: 1) organizing a bipolar spindle formation and 2) inducing cortical polarization and assembly of a distinct cortical cytoskeleton structure in the overlying cortex for polar body extrusion. At fertilization, a sperm brings exogenous paternal chromatin into the egg, which induces ectopic cortical polarization at the sperm entry site and leads to a cone formation, known as fertilization cone. Here we show that the sperm chromatin-induced fertilization cone formation is an abortive polar body extrusion due to lack of spindle induction by the sperm chromatin during fertilization. If experimentally manipulating the fertilization process to allow sperm chromatin to induce both cortical polarization and spindle formation, the fertilization cone can be converted into polar body extrusion. This suggests that sperm chromatin is also able to induce polar body extrusion, like its maternal counterpart. The usually observed cone formation instead of ectopic polar body extrusion induced by sperm chromatin during fertilization is due to special sperm chromatin compaction which restrains it from rapid spindle induction and therefore provides a protective mechanism to prevent a possible paternal genome loss during ectopic polar body extrusion
Freeze-dried primate sperm retains early reproductive potential after intracytoplasmic sperm injection
Oocytes inseminated by intracytoplasmic sperm injection using fresh ejaculated or freeze-dried rhesus macaque sperm showed similar activation, sperm aster assembly, and male-female pronuclear apposition rates.http://www.sciencedirect.com/science/article/B6T6K-4NY4Y3G-3/1/b88ff573caaf542e4e022e698231d4b
Ex Vivo Reconstitution of Arterial Endothelium by Embryonic Stem Cell-Derived Endothelial Progenitor Cells in Baboons
SNAREs in Mammalian Sperm: Possible Implications for Fertilization
Soluble N-ethylmalameide-sensitive factor attachment protein receptor (SNARE) proteins are present in mammalian sperm and could be involved in critical membrane fusion events during fertilization, namely the acrosome reaction. Vesicle-associated membrane protein/synaptobrevin, a SNARE on the membrane of a vesicular carrier, and syntaxin 1, a SNARE on the target membrane, as well as the calcium sensor synaptotagmin I, are present in the acrosome of mammalian sperm (human, rhesus monkey, bull, hamster, mouse). Sperm SNAREs are sloughed off during the acrosome reaction, paralleling the release of sperm membrane vesicles and acrosomal contents, and SNARE antibodies inhibit both the acrosome reaction and fertilization, without inhibiting sperm-egg binding. In addition, sperm SNAREs may be responsible, together with other sperm components, for the asynchronous male DNA decondensation that occurs following intracytoplasmic sperm injection, an assisted reproduction technique that bypasses normal sperm-egg surface interactions. The results suggest the participation of sperm SNAREs during membrane fusion events at fertilization in mammals.http://www.sciencedirect.com/science/article/B6WDG-45F53DF-6N/1/049af5c994522859c35001d18bc0f94
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Direct Differentiation of Human Pluripotent Stem Cells into Haploid Spermatogenic Cells
Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) have been shown to differentiate into primordial germ cells (PGCs) but not into spermatogonia, haploid spermatocytes, or spermatids. Here, we show that hESCs and hiPSCs differentiate directly into advanced male germ cell lineages, including postmeiotic, spermatid-like cells, in vitro without genetic manipulation. Furthermore, our procedure mirrors spermatogenesis in vivo by differentiating PSCs into UTF1-, PLZF-, and CDH1-positive spermatogonia-like cells; HIWI- and HILI-positive spermatocyte-like cells; and haploid cells expressing acrosin, transition protein 1, and protamine 1 (proteins that are uniquely found in spermatids and/or sperm). These spermatids show uniparental genomic imprints similar to those of human sperm on two loci: H19 and IGF2. These results demonstrate that male PSCs have the ability to differentiate directly into advanced germ cell lineages and may represent a novel strategy for studying spermatogenesis in vitro
Direct Differentiation of Human Pluripotent Stem Cells into Haploid Spermatogenic Cells
Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) have been shown to differentiate into primordial germ cells (PGCs) but not into spermatogonia, haploid spermatocytes, or spermatids. Here, we show that hESCs and hiPSCs differentiate directly into advanced male germ cell lineages, including postmeiotic, spermatid-like cells, in vitro without genetic manipulation. Furthermore, our procedure mirrors spermatogenesis in vivo by differentiating PSCs into UTF1-, PLZF-, and CDH1-positive spermatogonia-like cells; HIWI- and HILI-positive spermatocyte-like cells; and haploid cells expressing acrosin, transition protein 1, and protamine 1 (proteins that are uniquely found in spermatids and/or sperm). These spermatids show uniparental genomic imprints similar to those of human sperm on two loci: H19 and IGF2. These results demonstrate that male PSCs have the ability to differentiate directly into advanced germ cell lineages and may represent a novel strategy for studying spermatogenesis in vitro