scholarly journals The course of the acrosome reaction in guinea-pig sperm

1982 ◽  
Vol 54 (1) ◽  
pp. 161-171
Author(s):  
D.P. Green
Keyword(s):  
Guinea Pig ◽  
Membrane Fusion ◽  
Osmotic Pressure ◽  
Acrosome Reaction ◽  
Plasma Membranes ◽  
Calcium Ionophore ◽  
Colloid Osmotic Pressure ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Sequence Of Events

The acrosome reaction in guinea-pig sperm is accompanied by a marked cavitation of the acrosomal contents. Two divergent views are held as to whether this cavitation precedes or follows the membrane fusion that occurs in the reaction. To distinguish between these 2 views cavitation was induced in media containing a colloid, either Ficoll 70 or inulin, either by inducing a normal acrosome reaction using the calcium ionophore A23187 or by using the detergent Triton X100. Both Ficoll 70 and inulin, when incorporated into media of normal osmolality, were able to suppress various features of the cavitation. Complete retention of acrosomal shape was achieved in sperm treated with detergent in 30% (W/V) Ficoll 70 solution despite the absence of the limiting acrosomal and plasma membranes. This evidence supports the suggestion that the cause of the cavitation is a colloid osmotic pressure within the acrosomal matrix. This in turn supports one of the 2 proposed mechanisms for the temporal sequence of events occurring in the acrosome reaction.

Relocation of myosin and actin, kinesin and tubulin in the acrosome reaction of bovine spermatozoa

2009 ◽  
Vol 21 (2) ◽  
pp. 364 ◽  
Author(s):  
Ifigenia Oikonomopoulou ◽  
Hitesh Patel ◽  
Paul F. Watson ◽  
Peter D. Chantler
Keyword(s):  
Molecular Motors ◽  
Acrosome Reaction ◽  
Molecular Motor ◽  
Calcium Ionophore ◽  
Cell Types ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Sequence Of Events ◽  
Bovine Spermatozoa ◽  
Number Of Cells

The mammalian acrosome reaction is a specialised exocytotic event. Although molecular motors are known to be involved in exocytosis in many cell types, their potential involvement in the acrosome reaction has remained unknown. Here, it has been shown that actin is localised within the equatorial segment and in the marginal acrosomal ridge of the heads of unreacted bull spermatozoa. Myosins IIA and IIB are found within the anterior acrosomal margins of virtually all sperm cells and, less prominently, within the equatorial segment. Tubulin was detected in the equatorial segment and around the periphery of the acrosome while kinesin was prominent in the equatorial segment. After induction of the acrosome reaction by means of the calcium ionophore A23187, the number of cells exhibiting actin fluorescence intensity in the anterior acrosomal margin decreased four-fold and those displaying equatorial segment fluorescence decreased 3.5-fold; myosin IIA immunofluorescence decreased in intensity with most spermatozoa losing equatorial staining, whereas there was little change in the distribution or intensity of myosin IIB immunofluorescence, except for a ~20% decrease in the number of cells exhibiting acrosomal staining. Tubulin became largely undetectable within the head and kinesin staining spread rostrally over the main acrosome region. A possible sequence of events that ties in these observations of molecular motor involvement with the known participation of SNARE proteins is provided.

Acrosome stability in the spermatozoa of dasyurid marsupials

2008 ◽  
Vol 20 (2) ◽  
pp. 295 ◽  
Author(s):  
N. A. Czarny ◽  
K. E. Mate ◽  
J. C. Rodger
Keyword(s):  
Disulfide Bonds ◽  
Acrosome Reaction ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Macropus Eugenii ◽  
Sminthopsis Crassicaudata ◽  
Triton X ◽  
Physical And Chemical ◽  
Physical Challenge

The spermatozoa of most marsupials lack nuclear stabilising disulfide-bonded protamines found in eutherian mammals. However, disulfide stabilisation has been observed in the acrosome of macropodid (Macropus eugenii) and phalangerid (Trichosurus vulpecula) marsupials. As a result this organelle, which is normally fragile in eutherian mammals, is robust and able to withstand physical and chemical challenge in these marsupials. The present study examined acrosomal characteristics of the spermatozoa of three dasyurid marsupials; the fat-tailed dunnart (Sminthopsis crassicaudata), eastern quoll (Dasyurus viverrinus) and northern quoll (Dasyurus hallucatus). In all species examined Bryan’s staining demonstrated that significant acrosomal loss occurred following physical challenge with osmotic stress, cryopreservation without cryoprotectant and exposure to detergent (Triton-X). Bromobimane staining indicated that the acrosomes of dasyurids lacked stabilising disulfide bonds. As reported for the wallaby and possum, calcium ionophore (A23187) did not induce the acrosome reaction-like exocytosis in dasyurid spermatozoa but treatment with diacylglycerol (DiC8) caused significant acrosome loss at concentrations similar to those effective for other marsupials. The present study found that the spermatozoa of dasyurids are more sensitive to physical challenge than the previously-studied marsupials and we suggest that this is due to the absence of acrosomal stabilising disulfide bonds.

Leptin is involved in acrosome reaction by facilitating activation of MAPK cascades in the Chinese mitten crab, Eriocheir sinensis

2020 ◽  
Vol 70 (1) ◽  
pp. 81-95
Author(s):  
Qing Li ◽  
Haitao Zhao ◽  
Lin He ◽  
Hongdan Yang ◽  
Qun Wang
Keyword(s):  
Acrosome Reaction ◽  
Calcium Ionophore ◽  
Eriocheir Sinensis ◽  
Calcium Ionophore A23187 ◽  
Mitogen Activated Protein Kinase ◽  
Ionophore A23187 ◽  
Mapk Cascades ◽  
Mitten Crab ◽  
Mitogen Activated Protein

Abstract The role of leptin has been documented in several studies, including activated threonine phosphorylation of extracellular signal-regulated kinase (ERK1/2) in the reproduction of rodents and humans. Our previous studies have demonstrated that mitogen-activated protein kinase (MAPK) cascades ERK, P38, and c-Jun N-terminal kinase (JNK) are involved in the spermatogenesis and acrosome reaction of Eriocheir sinensis. Therefore, the aim of this study was to investigate the expression of leptin and its receptor (LepR), and the effect of leptin on MAPK cascades during calcium ionophore A23187-induced spermatozoa acrosome reaction in crabs. Successful western blotting revealed a 16 kDa band for leptin, and 120 kDa and 90 kDa bands for the obese receptor (LepR), respectively, in the tested male reproductive tissues. Both leptin and LepR were localized at the pro-acrosomal vesicle and apical cap (AC) of spermatids, suggesting their role in the subsequent acrosome reaction. Moreover, acrosome reaction can be enhanced by leptin, and this effect decreased due to the anti-LepR antibody. Afterwards, we investigated the effects of leptin on MAPK cascades. The results showed that leptin mainly activated the phosphorylation of ERK, P38 and JNK proteins in the apical cap during the acrosome reaction in crab spermatozoa. This study addresses the role of leptin on spermatozoa, and suggests that leptin may induce molecular changes associated with spermatozoa during acrosome reaction.

scholarly journals A role for protein kinase C in the membrane fusion necessary for platelet granule secretion

Blood ◽  
1989 ◽  
Vol 74 (7) ◽  
pp. 2405-2413 ◽  
Author(s):  
JM Gerrard ◽  
LL Beattie ◽  
J Park ◽  
SJ Israels ◽  
A McNicol ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Membrane Fusion ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ultrastructural Changes ◽  
Ionophore A23187 ◽  
Kinase C ◽  
Granule Secretion ◽  
Dimethyl Amiloride

Abstract The addition of 1-oleoyl-2-acetylglycerol (OAG), or phorbol-12- myristate-13-acetate (PMA) to platelets induced the phosphorylation of a 47,000 dalton protein (47 Kd), fusion of granule membranes with membranes of the surface connected canalicular system, the formation of large vesicles and the secretion of serotonin. 1-(5- isoquinolinesulfonyl)-2-methyl-piperazine (H7), and sphingosine, inhibitors of protein kinase C, significantly inhibited the ultrastructural changes and the phosphorylation of 47 Kd. N-(2- guanidinoethyl)-5-isoquinolinesulfonamide (HA1004), structurally similar to H7, but a weaker inhibitor of protein kinase C, did not attenuate these responses to OAG or to PMA. H7, but not HA1004, also markedly inhibited secretion induced by the synergistic combination of OAG and the calcium ionophore A23187. Amiloride and 5-(N,N dimethyl)- amiloride, inhibitors of the Na+/H+ transporter, did not inhibit the ultrastructural response and the protein phosphorylation induced by OAG, or the secretion induced by the combination of A23187 and OAG. The results link the activation of protein kinase C by diglycerides to the labilization and fusion of granule membranes important for secretion.

scholarly journals Cross-regulation of cortisol secretion by adrenocorticotropin and platelet-activating factor in perfused guinea pig adrenals

2007 ◽  
Vol 195 (1) ◽  
pp. 29-38
Author(s):  
Toshio Shimada ◽  
Taeko Hirose ◽  
Itsuro Matsumoto ◽  
Tadaomi Aikawa
Keyword(s):  
Protein Kinase ◽  
Guinea Pig ◽  
Platelet Activating Factor ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Cortisol Response ◽  
Cortisol Secretion ◽  
Induced Signal ◽  
Dependent Processes

We examined the cross-regulation of signaling between ACTH-and platelet-activating factor (PAF)-mediated steroidogenesis in the perfused guinea pig adrenal gland. Our method of in situ perfusion using an artificial medium can evaluate whether cortisol secretion in response to ACTH and PAF is interactive. Treating adrenal glands with 100 pg/ml ACTH diminished the subsequent cortisol response to 10 nM PAF. By contrast, PAF resulted in subsequent potentiation of ACTH-induced cortisol secretion. A mixture of 50 μM l-α-1-oleoyl-2-acetyl-sn-glycerol (OAG), an activator of protein kinase C (PKC), and 3.3 μM calcium ionophore (A23187), or 10 μM forskolin (FRK) diminished the cortisol response to PAF, whereas that to ACTH was unaffected. Each of PAF, ACTH, or FRK eliminated the cortisol response to OAG plus A23187, whereas that to FRK was unaffected. These data show that the protein kinase A (PKA)-dependent processes activated by ACTH or FRK can interfere with PAF-induced signal transduction at receptor and post-receptor levels. In contrast, PKC-dependent processes activated by PAF promoted ACTH-signaling at receptor and post-receptor level. Cross-regulation between processes activated by PAF receptor–PKC and by ACTH receptor–PKA might function in the multifactorial regulation of adrenocortical steroidogenesis.

In vitro induction of the acrosome reaction in ovine spermatozoa by calcium ionophore A23187

2003 ◽  
Vol 51 (1) ◽  
pp. 103-109 ◽  
Author(s):  
Ö. Uçar ◽  
T. J. Parkinson
Keyword(s):  
Phase Contrast ◽  
Acrosome Reaction ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Contrast Microscopy ◽  
The Relationship ◽  
In Vitro Induction ◽  
Artificial Vagina

The relationship between concentration of calcium ionophore A23187 and incubation time upon the proportion of spermatozoa undergoing acrosome reaction (AR) in vitro was investigated in rams from a commercial artificial insemination (AI) program. Two ejaculates were collected by artificial vagina from each of nine rams of three breeds (Finn Dorset, Charolais and Suffolk) aged 8-36months. Each ejaculate was diluted in a skimmed milk extender. Spermatozoa were thereafter incubated for 45 or 60min in modified Tyrode's medium (TALP) which contained either zero, 0.1 or 1.0µM/l A23187. After fixing in 10% formaldehyde, the number of spermatozoa that had undergone AR was determined by phase contrast microscopy. In pre-incubation samples, 21.3± 3.3% of spermatozoa had undergone AR. Percentages of acrosome reacted spermatozoa were significantly (P<0.001) increased after incubation with A23187. After incubation with 0.1µM/l A23187 for 45 and 60min there were 22.4±3.0% and 31.7±4.3% acrosome reacted spermatozoa, respectively. After incubation with 1.0µM/l A23187 for 45 and 60min there were 46.2±6.5% and 53.8±5.9% acrosome reacted spermatozoa, whilst corresponding numbers in control samples were 17.0±2.7% and 22.3±4.2%. There was also a significant (P<0.001) effect of individual animals upon the responses to different concentrations of A23187. These findings indicate that (i) A23187 can be used to assess the AR of ovine spermatozoa in vitro and (ii) there are effects of individual animals upon the proportion of spermatozoa undergoing AR.

The calcium ionophore A23187 stimulates glycoprotein secretion by the guinea pig gallbladder

Hepatology ◽  
1986 ◽  
Vol 6 (4) ◽  
pp. 569-573 ◽  
Author(s):  
Peter F. Malet ◽  
Catherine L. Locke ◽  
Bruce W. Trotman ◽  
Roger D. Soloway
Keyword(s):  
Guinea Pig ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Glycoprotein Secretion
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