Sperm chemotaxis is a chemical guiding mechanism that may orient spermatozoa to the egg surface. A picomolar concentration gradient of Progesterone (P), the main steroidal component secreted by the cumulus cells that surround the egg, attracts human spermatozoa. In order to elucidate the molecular mechanism of sperm chemotaxis mediated by P, we combine the application of different strategies: pharmacological inhibition of signaling molecules, measurements of the concentrations of second messengers and activation of the chemotactic signaling. Our data implicate a number of classic signal transduction pathways in the response and provide a model for the sequence of events, where the tmAC-cAMP-PKA pathway is activated first, followed by protein tyrosine phosphorylation (equatorial band and flagellum) and calcium mobilization (through IP3R and SOC channels), whereas the sGC-cGMP-PKG cascade, is activated later. These events lead to sperm orientation towards the source of the chemoattractant. The finding proposes a molecular mechanism which contributes to the understanding of the signal transduction pathway that takes place in a physiological process as chemotaxis.
Sperm chemotaxis in mammals have been identified towards several female sources as follicular fluid (FF), oviduct fluid, and conditioned medium from the cumulus oophorus (CU) and the oocyte (O). Though several substances were confirmed as sperm chemoattractant, Progesterone (P) seems to be the best chemoattractant candidate, because: 1) spermatozoa express a cell surface P receptor, 2) capacitated spermatozoa are chemotactically attracted in vitro by gradients of low quantities of P; 3) the CU cells produce and secrete P after ovulation; 4) a gradient of P may be kept stable along the CU; and 5) the most probable site for sperm chemotaxis in vivo could be near and/or inside the CU. The aim of this study was to verify whether P is the sperm chemoattractant secreted by the rabbit oocyte-cumulus complex (OCC) in the rabbit, as a mammalian animal model. By means of videomicroscopy and computer image analysis we observed that only the CU are a stable source of sperm attractants. The CU produce and secrete P since the hormone was localized inside these cells by immunocytochemistry and in the conditioned medium by enzyme immunoassay. In addition, rabbit spermatozoa express a cell surface P receptor detected by western blot and localized over the acrosomal region by immunocytochemistry. To confirm that P is the sperm chemoattractant secreted by the CU, the sperm chemotactic response towards the OCC conditioned medium was inhibited by three different approaches: P from the OCC conditioned medium was removed with an anti-P antibody, the attractant gradient of the OCC conditioned medium was disrupted by a P counter gradient, and the sperm P receptor was blocked with a specific antibody. We concluded that only the CU but not the oocyte secretes P, and the latter chemoattract spermatozoa by means of a cell surface receptor. Our findings may be of interest in assisted reproduction procedures in humans, animals of economic importance and endangered species.
CRISP1 is expressed by cumulus cells and plays a role in fertilization by modulating sperm orientation, hyperactivation, and key Ca2+ channels in sperm.
The guidance of human sperm cells under confinement in quasi 2D microchambers is investigated using a purely physical method to control their distribution. Transport property measurements and simulations are performed with dilute sperm populations, for which effects of geometrical guidance and concentration are studied in detail. In particular, a trapping transition at convex angular wall features is identified and analyzed. We also show that highly efficient microratchets can be fabricated by using curved asymmetric obstacles to take advantage of the spermatozoa specific swimming strategy.PACS numbers: 87.17. Jj, 87.18.Hf, 87.17.Aa, 87.17.Rt Understanding sperm dynamics under confining microgeometries is a general problem and a major challenge both from the basic biophysics and the complex fluids points of view. It is also crucial for microfluidics and biomedical control applications. Our knowledge of the swimming cell motilities in unbounded media cannot be directly extrapolated to their behavior in complex environments such as those found in the oviduct or in the labon-a-chip microfluidic devices used to control and analyze small samples or for in-vitro reproduction procedures. In these cases, the characteristic length scales are of the same order as the cell size, i.e. a few micrometers. Under these circumstances, confined self-propelled microorganisms undergo substantial changes in their locomotion habits, adapting their dynamics to intricate porous media or to solid surfaces vicinity [1], reducing their speed close to boundaries [2] or adjusting their morphology and motility in very narrow channels [3].It has been shown that microswimmers with very different propulsion systems are similarly attracted to the walls and to swim parallel to the surface [2,[4][5][6][7][8][9][10][11]. It is believed that this attractive force has hydrodynamic origin although other possible mechanisms have been proposed [12][13][14]. Several models have been introduced to describe the swimming along surfaces (see Ref.[15] and references therein). Interestingly, the direct observation of the cell-wall attraction (see Fig. 1) have led to the design of ratchet devices that guide and sort self-propelled cells using asymmetric obstacles [16,17]. In particular, different microfluidic devices have been created to either increase sperm cell quality or enhance their concentration [18][19][20]. The creation of inhomogeneous distributions of swimmer populations via asymmetric obstacles has been shown to be particularly efficient for run-and-tumble bacteria [16,[21][22][23]. Alternative ways of achieving nonuniform distributions have also been obtained combining symmetric funnels and flux [24]. Nowadays, numerous theoretical treatments are available to account for the effects of asymmetric obstacles on active particles distributions [25][26][27][28]. Tumbles, rotational diffusion and collisions are efficient mechanisms for separating the cells from the surface, thus permitting bacteria to be reinserted into the bulk of the confining micro...
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