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1.
The medaka, Oryzias latipes, is a well-recognized fish model for biomedical research. An understanding of gamete characteristics is necessary for experimental manipulations such as artificial fertilization and sperm cryopreservation. The goal of this study was to investigate sperm characteristics of motility initiation, duration, and retention in medaka. First, motility was initiated by osmolality values ranging from 25 to 686 mOsm/kg, which included deionized water and hypotonic, isotonic, and hypertonic Hanks’ balanced salt solution. The percentage of motile sperm was >80% when osmolality was <315 mOsm/kg and decreased as osmolality increased. This is different from most fish with external fertilization in which sperm motility can be initiated by hypotonic (for freshwater fish) or hypertonic (for marine fish) solutions or by altering the concentration of specific ions such as potassium (e.g., in salmonids). Second, upon activation, the sperm remained continuously motile, with reserve capacity, for as long as 1 wk during storage at 4 °C. This was also different from other externally fertilizing fish, in which motility is typically maintained for seconds to several minutes. Third, after changing the osmolality to 46 to 68 mOsm/kg by adding deionized water, the motility of sperm held at 274 to 500 mOsm/kg was higher than the original motility (P ≤ 0.035) after 24, 48, and 72 h of storage at 4 °C. Fourth, the addition of glucose had no effect on maintaining sperm motility during refrigerated storage. To our knowledge, this combination of sperm motility characteristics is reported for the first time in fish and may be unique to medaka or may represent an undescribed modality of sperm behavior within euryhaline fish.  相似文献   

2.
Limulus spermatozoa are nonmotile when spawned and become motile only after encountering a sperm motility initiating factor (SMI) exuded by the egg. SMI extracts (produced by washing intact eggs with distilled water, lyophilizing the supernatant to dryness, and redissolving the dried extract in artificial seawater, ASW) initiate sperm motility in the absence of eggs. Utilizing such SMI extracts, sperm motility initiation was found to be unaffected by changes in temperature from 16 to 30°C, pH from 6.3 to 8.6, and salinity from 85 to 125% ASW. Within these ranges, sperm motility initiation was an “all-or-nothing” response, with greater than 99% of the spermatozoa becoming motile. Also, each sperm swam with apparently the same speed (at a given temperature) until spontaneously stopping within 10 min after the addition of SMI extracts. Evidence was found that SMI may bind irreversibly to a receptor, which is inactivated within a few seconds or minutes, leading to the observed cessation of motility. Observations of sperm behavior near intact eggs showed no evidence of chemotaxis. Spermatozoa observed to swim toward intact eggs progressed with a uniform speed and were motile less than 5 sec from initiation of motility until attaching to the egg. The presence of an all-or-nothing response to SMI, the independence of sperm motility to experimental parameters, and several other characteristics of the animal and its spermatozoa make Limulus a potentially excellent model animal for examination of sperm motility control mechanisms.  相似文献   

3.
Synopsis Sea catfish, tested individually for 3-day periods in an automated data acquisition system, displayed a nocturnal pattern of locomotor activity with 62% of the activity occurring during scotophase. In addition, there were two relative maxima of activity at 0700–0900 and 1300–1500 C.S.T. This diel pattern of activity could not be correlated with tidal phenomena in the fish's natural environment and is virtually identical to that exhibited by several freshwater catfish species. Thus, even though sea catfish are a primarily estuarine fish and subject to daily tidal fluctuations, tidal rhythmicity is apparently not used as a cue for diel activity patterns.  相似文献   

4.
BACKGROUND INFORMATION: ATP is released from many cell types exposed to hypo-osmotic shock and is involved in RVD (regulatory volume decrease). Purinergic signalling events have been extensively investigated in mammals, but not in marine teleosteans. RESULTS: The effect of hypo-osmotic shock on ATP release was examined in isolated hepatocytes from turbot (Scophthalmus maximus), a marine flatfish. Hypo-osmotic stress (240 mOsm x kg(-1)) induced a significant increase in ATP efflux, and was inhibited by a potential CFTR (cystic fibrosis transmembrane conductance regulator) inhibitor, glibenclamide, but not by the MDR1 (multidrug resistance 1) P-glycoprotein inhibitor, verapamil. ATP efflux could be a cAMP-dependent process, as IBMX (isobutylmethylxanthine) and forskolin triggered the process under iso-osmotic conditions. Protein kinases, including protein kinase C, could also be involved, as staurosporine and chelerythrine inhibited the mechanism. Calcium could contribute to ATP efflux as ionomycin, a calcium ionophore, elicited a rapid release under iso-osmotic conditions, and chelation using EGTA abolished ATP release under hypo-osmotic conditions. RVD was partially abolished by apyrase, an ATP scavenger, and suramin, a purinoceptor antagonist. Moreover, hypo-osmotic shock induced a rise in intracellular calcium which could be involved in RVD. Since extracellular ATP triggered an increase in cellular free-calcium content under iso-osmotic conditions, our results could indicate that hypo-osmotic-induced ATP efflux contributes to RVD in turbot hepatocytes by stimulating purinergic receptors, which may lead to activation of a calcium signalling pathway. CONCLUSIONS: These data provide the first evidence of volume-sensitive ATP signalling for volume maintenance in a marine teleost fish cell type.  相似文献   

5.
The aims of this study were to test the presence of discrete sperm subpopulations in Florida goat ejaculates using a computer-assisted sperm analysis (CASA) system and to establish the relationship between the distribution of the subpopulations found and individual buck, total motility, and sperm concentration. Clustering methods and discriminant analysis were applied to identify motile sperm subpopulations within the semen samples. Principal component analysis revealed that three principal components represented more than the 88% of the variance. After the cluster analysis was performed four motile sperm subpopulations were identified. Subpopulation 1 consisted of rapid and linear sperm (39.84%), Subpopulation 2 consisted of slow but linear spermatozoa (33.23%), Subpopulation 3 consisted of rapid, high ALH but non-linear spermatozoa (14.63%), and Subpopulation 4 consisted of slow and non-linear spermatozoa (12.31%). There were significant differences in the distribution of the four subpopulations (P < 0.001) as well as in the percentage of total motility and the overall sperm concentration (P < 0.05) in fresh ejaculates among the four bucks tested. In conclusion, four well-defined motile sperm subpopulations were identified in Florida goat ejaculates. The relationship between the distribution of the sperm subpopulations and individual buck, total motility, and sperm concentration shows that the spermatozoa of each have different motility patterns. Therefore, the study of discrete subpopulations of motile spermatozoa could lead to a substantial increase in information acquired during caprine semen analysis.  相似文献   

6.
The efficiencies of water absorption from the guts of the larvae of herring ( Clupea harengus L.) and turbot ( Scophthalmus maximus L.) were estimated by two methods, The first method was based on the differences in the rates of accumulation, by drinking, and clearance from the gut of radiolabelled inert markers. The second method used the equilibrium level of radioactivity in the larvae to measure the concentration of the markers in the gut above background as water is absorbed from the gut. Water absorption efficiencies for herring larvae drinking sea water were estimated to be 77% using both methods. When external salinity was reduced to 50% sea water, drinking rates and water absorption efficiency in herring larvae fell substantially.
Estimates of water absorption efficiency of seawater-adapted turbot larvae were similar (71–84%) to that of herring using both methods. Although temperature had a marked effect on both the rate of drinking and water absorption, there was no significant thermal effect on the efficiency of absorption from the guts of turbot larvae. The limitations of the techniques and the implications of the estimates in terms of water balance in fish larvae are discussed.  相似文献   

7.
Sperm motility in fishes. (II) Effects of ions and osmolality: a review   总被引:2,自引:0,他引:2  
The spermatozoa of most fish species are immotile in the testis and seminal plasma. Therefore, motility is induced after the spermatozoa are released into the aqueous environment during natural reproduction or into the diluent during artificial reproduction. There are clear relationships between seminal plasma composition and osmolality and the duration of fish sperm motility. Various parameters such as ion concentrations (K+, Na+, and Ca2+), osmotic pressure, pH, temperature and dilution rate affect motility. In the present paper, we review the roles of these ions on sperm motility in Salmonidae, Cyprinidae, Acipenseridae and marine fishes, and their relationship with seminal plasma composition. Results in the literature show that: 1. K+ is a key ion controlling sperm motility in Salmonidae and Acipenseridae in combination with osmotic pressure; this control is more simple in other fish species: sperm motility is prevented when the osmotic pressure is high (Cyprinidae) or low (marine fishes) compared to that of the seminal fluid. 2. Cations (mostly divalent, such as Ca2+) are antagonistic with the inhibitory effect of K+ on sperm motility. 3. In many species, Ca2+ influx and K+ or Na+ efflux through specific ionic channels change the membrane potential and eventually lead to an increase in cAMP concentration in the cell, which constitutes the initiation signal for sperm motility in Salmonidae. 4. Media that are hyper- and hypo-osmotic relative to seminal fluid trigger sperm motility in marine and freshwater fishes, respectively. 5. The motility of fish spermatozoa is controlled through their sensitivity to osmolality and ion concentrations. This phenomenon is related to ionic channel activities in the membrane and governs the motility mechanisms of axonemes.  相似文献   

8.
Upon dilution into sea water, Limulus spermatozoa undergo a brief flurry of motility (duration < 60 sec), after which they are nonmotile until encountering a sperm motility initiating peptide (SMI) that emanates from eggs. Utilizing highly purified SMI extracts and simplified seawater formulations (from which individual ions have been deleted), we found that no specific extracellular ion is required for either dilution-initiated or SMI-initiated motility. Indeed, deletion of one ion (Na+) produced dilution-initiated motility of very long duration (several hours). When motility is initiated by SMI (in normal seawater) there is an increase in intracellular pH (pHi), as indicated by the fluorescent probe, 9-amino acridine; however, this pH, change is not a trigger for motility. As a more general method examining ion movements, the fluorescent probe diS-C3-(5) was used to qualitatively measure changes in the membrane potential of spermatozoa. Although crude SMI extracts caused membrane depolarization, further purification resulted in an almost complete separation of this activity from SMI, thus showing that SMI activation is apparently an electroneutral event. (The membrane-depolarizing factor has a molecular weight > 30,000 and does not initiate acrosome reactions.) Experiments utilizing the ionophore A23187 and Ca+2-blocking agents (verapamil and TMB-8) provided tentative evidence that mobilization of intracellular Ca+2 may be required for motility initiation. These results show that neither changes in pHi nor the influx of specific extracellular ions are direct mediators of SMI-initiated motility; however, experiments with pharmacologic agents indicate a possible role for intracellular Ca+2.  相似文献   

9.
The aim of this study was to investigate ageing of ovulated eggs retained in the ovarian lumen of captive turbot, Scophthalmus maximus L., prior to handstripping. The ovulation times of egg-batches were determined by catheterization, handstripping and plotting the percentage fertilizations and hatches of resulting eggs against time. The catheterization experiments showed that eggs age and change appearance rapidly after ovulation. The stripping method demonstrated that freshly-ovulated eggs showed greater than 90% fertilization and up to 97% hatch, but ovulated eggs retained in the ovary lumen for 1 day before stripping gave 0% hatch. The turbot showed precise ovulatory rhythms. The time between ovulations varied between individuals but was constant for any one female, making it possible to predict future ovulation times. Three of the females studied had ovulatory periods of alternating length.  相似文献   

10.
The presence and localization of acid and alkaline phosphatase, non-specific proteases, aminopeptidase, amylase, non-specific esterase and lipase was investigated by histoenzymologic methods in fed and fasting turbot from day 1 to day 40 post-hatching and compared with published data. Alkaline phosphatase and aminopeptidase activities were delected at day 1 in the distal region of the developing digestive tube. At day 3 (opening of the mouth) aminopeptidase and alkaline phosphatase activities were found all along the intestine. Sites of non-specific esterase and protease activities became apparent in the digestive tract at days 2 and 3 respectively. Amylase was present in the exocrine pancreas at day 3 and in the lumen of the intestine at day 4. Acid phosphatase was active in the cellular structure surrounding the yolk stores and in the lipid droplets at day 1 and in the intestinal epithelium at day 3. Lipase was found at day 15 when the larvae metamorphose into juveniles.
All the investigated enzymes were detected in fasting animals, except for lipase. However, the intensities of the enzymatic activities were weaker in the fasting specimens relative to the fed specimens between days 7 and 10.  相似文献   

11.
Egg extracts (obtained by washing intact Limulus eggs with either distilled water or artificial seawater, ASW) contain a sperm motility initiating factor (SMI). The SMI is heat stable (withstands boiling to dryness), passes through a dialysis membrane, and is retained by G-10 Sephadex (indicating a molecular weight of less than 700). Qualitative analysis (by X-ray fluorescence spectroscopy) and quantitative analysis (by atomic absorption spectroscopy) of SMI extracts revealed the presence of four divalent cations (Ca, Mg, Ni, and Cu) and one monovalent cation (K) that affect sperm motility. When assayed individually at high concentrations, all of the divalent cations initiate sperm motility and K+ inhibits motility initiation by the divalent cations. However, none of the divalent cations are present at concentrations high enough to produce the observed SMI activity, and since K+ is present when motility is initiated by SMI, K+ must not normally be an inhibitor. Therefore, if inorganic cations are involved in normal sperm motility initiation in Limulus, they are acting in conjunction with some other low molecular weight factor.  相似文献   

12.
大菱鲆仔稚幼鱼消化系统发育的组织学研究   总被引:9,自引:0,他引:9  
大菱鲆(Scophthalmus maximus) 是目前欧洲重要的海水养殖良种之一, 在中国北方沿海大菱鲆的养殖规模也在不断扩大。    相似文献   

13.
In Vimba vimba, GSI, sperm volume, and spermatozoa concentration range from 3.4-7.4 %, 0.1-1.1 ml, and 13.3-34.8 × 109 spz ml−1, respectively. Gonad mass (r = 0.90) and sperm volume (r = 0.35) significantly correlated with weight of males. Significant correlation was also found between gonad mass and length of males (r = 0.85). Sperm motility (r = 0.99) and velocity (r = 098) significantly decreased after activation in Tris-HCl 20 mM, pH 8.5. Osmolality of the seminal plasma was 273.2 mOsmol kg−1. Sperm motility and velocity were significantly affected by the osmolality of the activation medium (P < 0.01). Hyper-osmolality compared to seminal plasma osmolality totally suppressed the sperm activation. At 15 s post-activation, the sperm motility significantly decreased at 240 mOsmol kg−1 in KCl or NaCl media. The highest sperm motility and velocity (at 60 s post-activation) were observed at 200 mOsmol kg−1 in NaCl, KCl, or sucrose media. In all treatments, the tip of the flagellum of spermatozoa became curled into a loop shape after activation of sperm in distilled water containing 20 mM Tris-HCl, pH 8.5 that shortened the flagellum.  相似文献   

14.
A protocol for storage of emu semen >6 h has not yet been optimized. The objective was to determine: a) whether sperm quality was adversely affected by sudden exposure to low temperatures (5, 10 and 20 °C) during collection; and b) the effects of three storage temperatures (5, 10 and 20 °C) on survival of emu sperm. In two experiments, each repeated three times on alternate days, ejaculates were diluted 1:1 with precooled (5, 10, or 20°C) UWA-E3 diluent and stored for up to 48 h. Collection temperature, or interaction with either the storage time or storage temperature, had no significant effect on sperm viability, motility, or morphology. Mass Motility Score (2.91-3.27 ± 0.26, mean ± SEM), and percentages of live (72.4-76.2 ± 2.4) and morphologically normal sperm (63.3-64.5 ± 2.3) were comparable among collection temperatures. Conversely, storage temperature and storage time affected (P < 0.05) sperm viability, motility, and morphology. After storage for 48 h, percentages of viable, normal, and motile sperm were higher (P < 0.001) at 5 °C (58.7% ± 1.1, 44.7% ± 1.3, and 50.7% ± 4.9, respectively) and 10 °C (62.6% ± 1.1, 54.1% ± 1.3, and 60.4% ± 4.9) than at 20 °C (27.6% ± 1.1, 20.1% ± 1.3, and 25.9% ± 4.9). Beyond 6 h of storage, the percentage of abnormal sperm was higher (P < 0.001) for storage at 5 °C compared to 10 and 20 °C. After 48 h, bacterial counts were considerably higher at 20 °C compared to 5 and 10 °C (P < 0.001). The pH of stored sperm suspension remained unaffected at 5 and 10 °C, but at 20 °C declined to 6.5 ± 0.03 after 24 h (P < 0.05) and to 6.0 ± 0.03 after 48 h (P < 0.001). We concluded that emu semen could be collected at low ambient temperatures (5-20 °C) without compromising its in vitro storage duration and that semen quality during storage for 48 h was better if it was stored at 10 °C than at 5 or 20 °C.  相似文献   

15.
Motile sperm cells of land plants are released directly into the environment and encounter numerous constraints on their way to the egg. Sperm cell organization, shape, size, and plasticity are crucial to the processes associated with fertilization. We conducted an ultrastructural investigation to detail insemination (sperm release, swimming and movement within the archegonium) and fertilization in the model fern Ceratopteris richardii. Gametophytes were grown from spores using sterile culture techniques and flooded in water when sexually mature. Materials were examined at different stages post-flooding. During insemination in C. richardii, the sperm cytoskeleton and flagella rearrange, and the coils of the cell extend while entering the neck canal. In this nearly linear configuration, the dense ridge, a densely compacted band of filaments presumed to be actin, expands to surround the leading edge of the sperm cell. This ridge fuses with the receptive site on the female gamete and is the sperm component that initiates contact with the egg nuclear envelope. All cellular components, except plastids, enter the egg cytoplasm. Sperm mitochondria are distinguishable from those of the egg because they are encased by two or three additional membranes and are sequestered from the zygote cytoplasm. During karyogamy, the sperm components, including the microtubule cytoskeleton (spline) and flagella, maintain their spatial integrity. Microtubules play key roles not only in sperm cell structure but also in facilitating karyogamy in this fern. After karyogamy is completed, microtubule arrays of the sperm cell and the components of the locomotory apparatus are disassembled. We provide the first demonstration of the likely involvement of sperm actin in egg penetration in land plants and new insights into the fate of paternal organelles. This study points to the roles sperm cell structure and dynamics play in the intricate processes of insemination and fertilization in land plants.  相似文献   

16.
Transferrin (Tf) is a major protein of carp (Cyprinus carpio) seminal plasma. Its relationship with milt quality is unknown. In this study, we sought to determine if Tf is polymorphic in carp seminal plasma and if this polymorphism is related to sperm motility characteristics. We screened males of purebred common carp line (Polish line R6) for Tf polymorphism in blood plasma. The majority of Tf genotypes represented only DD and DG variants. We then collected milt from preselected DD and DG genotypes and tested their sperm motility characteristics using computer-aided sperm analysis (CASA). Tf polymorphism in seminal plasma was found to be identical with that of blood. However, the relationships between Tf polymorphism and iron metabolic parameters were different for blood and semen. These data suggest different regulation of Tf in liver and testis. We found substantial differences in sperm motility characteristics between both genotypes. Spermatozoa of DG males were characterized by lower curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), higher linearity (LIN) and straightness (STR) of movement as compared to DD males. No differences were found in other sperm characteristics such as sperm concentration and percentage of sperm motility. Our results suggest that sperm motility parameters are related to Tf polymorphism and therefore this polymorphism may be related to sperm competitive ability.  相似文献   

17.
The growth potential of turbot Scophthalmus maximus larvae and juveniles was studied using nucleic acid‐based indices and protein variables. The experiment was carried out from 4 to 60 days post hatching (dph). A significant increase in instantaneous growth rate during metamorphosis and retarded growth rate during post‐metamorphic phase were observed. Ontogenetic patterns of DNA, RNA and protein all showed developmental stage‐specific traits. The RNA:DNA ratio decreased up to 12 dph, then increased rapidly till 19 dph and fluctuated until 35 dph followed by a decline to the end. The RNA:DNA ratio was positively correlated with growth rate of juveniles during the post‐metamorphic phase, whereas this ratio was not a sensitive indicator of growth during the pre‐metamorphic phase and metamorphosis. The protein:DNA ratio showed a similar tendency to the RNA:DNA ratio. Changes of DNA content and protein:DNA ratio revealed that growth of S. maximus performed mainly by hyperplasia from 4 to 12 dph and hypertrophy until 21 dph during the pre‐metamorphic larval phase. Growth was dominantly hypertrophical from the early‐ to mid‐metamorphosing phase and hyperplastic thereafter. The results show that the DNA content and protein:DNA ratio can evaluate growth rates of larval and juvenile S. maximus on a cellular level.  相似文献   

18.
Despite the prevalence of zebrafish as a model scientific organism, understanding sperm function in this species is essentially limited to observations that osmotic shock initiates motility. During natural spawning, sperm encounter a range of environmental salinities as well as freshwater mixed with egg-associated ovarian fluid (OF), thus sperm are likely to be exposed to saline prior to egg contact. Effects of saline on sperm function in this model species are unknown, but likely to be important. Using computer assisted sperm analysis, this study addressed the effects of osmolality of spawning media and ionic composition and pH on the proportion of sperm becoming motile at activation (motility), as well as sperm velocity and path. When activated with tap water, motility was maximal (80%) at 10 s (earliest time measured), declining to 5% by 87 s postactivation. With activation at moderate osmolalities (∼160-200 mmol/kg) initial motility was decreased relative to low osmolality, increased from 10 to 30 s, and subsequently declined less rapidly (motility in 80 mM NaCl was 35%, 80%, and 60% at 10, 30 and 147 s, respectively). Thus, moderate osmolality increased duration, but introduced a temporal lag in motility onset. With moderate osmolalities, the rate of velocity decay was less than that with tap water activation. Sodium chloride and sucrose similarly impacted both motility and velocity. Replacement of NaCl with KCl, pH values ranging from 6.8 to 8.4, or the presence of gadolinium were without effect. Motility, but not velocity, was slightly supressed by Ca2+. Therefore, whereas pH and concentrations of Ca2+ or K+ of OF are unlikely to impact fertility via sperm motility, the OF contribution to spawning media osmolality may have pronounced effects on motility and velocity of sperm, factors previously correlated with fertility in other species.  相似文献   

19.
1. The effects of riffles as barriers to movement of stream fish was investigated in a set of eight large outdoor artificial streams. Pools were 183 cm in diameter and 45 cm deep; riffles were 183 cm long and 43 cm wide. Rates of movement of three species of minnows (Cyprinidae) (Campostoma anomalum, Cyprinella venusta and Notropis boops) among pools were measured at four riffle current velocities (0, 15, 30 and 45 cm s–1), three thalweg depths (10, 50 and 100 mm), two riffle lengths (183 and 549 cm), and with and without the threat of predation. Visual observation and video cameras were used to quantify movement rate. 2. Mean movement rate (percentage of fish crossing a riffle each 30 min) was 18.1% at 0 cm s–1 and only 1.8 at 45 cm s–1. Movement rate was 7.2% with no predators present and 20.2% with caged predators in pools. Notropis boops had a lower rate of movement than C. venusta or C. anomalum across all trials. The mean group size (number of individuals crossing a riffle together) was 1.2 fish overall, indicating most movement was by individuals and not groups. Group size was significantly greater only with shallow riffles or under the threat of predation. 3. Overall biotic and abiotic factors in these artificial streams do influence movement rates and may affect movement among pools in natural streams.  相似文献   

20.
虾蟹精子发育的营养及代谢研究概况   总被引:2,自引:0,他引:2  
对雄性虾蟹的生殖营养及精子代谢作了综述。大量研究证实,在人工养殖条件下,雄性虾蟹的生殖力会出现明显下降,其原因与人工饲养条件下营养缺乏有关。精子储存在贮精囊和纳精囊中时,不断进行旺盛的代谢活动,其代谢类型主要是无氧代谢,而代谢的能量物质大部分种类为碳水化合物,这些物质主要由精子以外的物质提供。  相似文献   

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