Change in intracellular K+ concentration caused by external osmolality change regulates sperm motility of marine and freshwater teleosts

1995 ◽  
Vol 108 (3) ◽  
pp. 1175-1181
Author(s):  
H. Takai ◽  
M. Morisawa

We previously demonstrated that osmolality isotonic to the seminal plasma suppresses sperm motility in marine and freshwater teleosts, and exposure of sperm to hypertonicity of sea water or hypotonicity of fresh water, respectively, induces the initiation of sperm motility at spawning. The motile sperm became immotile by return of osmolality to the isotonic osmolality both in a marine teleost, the puffer fish, and a freshwater teleost, the zebrafish. The initiation and termination of sperm motility could be repeated several times by changing surrounding osmolality in both species. In demembranated sperm, motility was suppressed by a K+ concentration equivalent to the seminal salt concentration in both puffer and zebrafish. Demembranated puffer sperm were reactivated when K+ concentration of the reactivating solution increased. Conversely, initiation of motility in the demembranated zebrafish sperm was induced by decreasing K+ concentration. The initiation and termination of the demembranated sperm were alternately repeated by changing K+ concentration of the reactivation solution in both species. Furthermore, intracellular K+ concentration rose when sperm motility of the puffer was initiated in hypertonic solutions. These results suggest that change in external osmolality is converted into change in intracellular K+ concentration, and that the change affects the flagellar axoneme as a signal to initiate or terminate sperm motility. The initiation and termination of motility in the demembranated puffer sperm were caused at a high pH and a low pH of the reactivating solution, respectively, suggesting the contribution of intracellular pH in the regulation of flagellar motility.

1974 ◽  
Vol 60 (3) ◽  
pp. 769-781
Author(s):  
T. J. SHUTTLEWORTH ◽  
R. F. H. FREEMAN

1. Measurements of net flux of water have been made on isolated gills removed from freshwater-adapted and seawater-adapted eels and incubated in various media of differing osmotic pressure. 2. From these measurements it has been possible to determine the osmotic permeability coefficient of the gill directly from the net water flux. The values obtained (0.50±0.14x10-5 cm.sec-1 for freshwater eels and 0.43±0.07x10-5 cm.sec-1 for seawater-adapted eels) indicate that there was no significant change in this parameter on adaptation of the eels to sea water. 3. The direct measurements made of the net water flux across the isolated gills appear to be compatible with the osmoregulatory pattern of eels as deduced by other workers using different techniques. In particular they illustrate and further emphasize the significance of drinking in the freshwater fish. 4. Calculations indicate that, for a freshwater teleost, the osmotic and ionic problems caused by drinking in fresh water have an insignificant energetic effect and hence, energetically, it matters little to the fish whether it drinks or not.


1971 ◽  
Vol 178 (1051) ◽  
pp. 151-160 ◽  

The spermatozoa of the giant octopus of the North Pacific, freshly removed from spermatophores, showed very little motility, but on dilution with sea-water or 2.7 % NaCl, followed by dialysis against either of these two media, they became intensely motile and remained so for several days at 2 to 10 °C. At higher temperatures, particularly above 25 °C, octopus spermatozoa lost their motility rapidly. At 35 °C, complete and irreversible loss of motility occurred within less than 1 min. The motility of octopus spermatozoa at 2 to 10 °C persisted under both anaerobic and aerobic conditions and did not require the presence of exogenous glycolysable sugar. The addition of spermatophoric plasma to a motile sperm suspension inhibited motility. Other inhibitors were sodium azide, 2, 4-dinitrophenol and ethylenediaminetetra-acetate, at 0.001 M concentrations. ATP, ADP and arginine phosphate have been identified and quantitatively measured in octopus spermatozoa. On prolonged incubation of motile sperm suspensions a t 3 °C, ATP and ADP did not decline appreciably, whilst arginine phosphate decreased considerably. The decrease was even more pronounced in sperm suspensions which had first been inactivated by short exposure to 35 °C, prior to prolonged incubation at 3 °C. Glycogen, the main carbohydrate store of octopus spermatozoa, remained at a high concentration even in sperm suspensions kept for 5 days at 3 °C, and there was no appreciable difference in that respect between a sample containing motile spermatozoa and one in which, at the outset of incubation, the spermatozoa were immobilized by heating to 35 °C.


2013 ◽  
Vol 13 (2) ◽  
Author(s):  
D. Setiadi Dan S. Bachri

AbstrakIndonesia memiliki banyak tumbuhan berpotensi obat salah satunya bisa dijadikan sebagai obatkontrasepsi tradisonal yang biasa digunakan untuk menjarangkan anak atau sterilisasi sepertirebusan akar Caesalpinia bonduc (Aru). Tujuan penelitian untuk mengetahui pengaruh obatkontrasepsi tradisional air rebusan akar aru terhadap kualitas pematangan sperma epididimis mencit(Mus musculus). Mencit dipilih secara acak untuk mewakili 4 kelompok dosis yaitu : 0,0 (K0), 16,0(K1) 24,0 (K2) dan 32,0 (K3) μl/gram berat badan/hari. Setiap kelompok perlakuan dilakukandengan 5 kali replikasi. Perlakuan diberikan melalui oral dengan menggunakan jarum gavage selama10 hari berturut-turut. Pembedahan dilakukan pada hari pertama setelah perlakuan selesai.Pengamatan kualitas sperma epididimis dengan perhitungan produksi sperma, % sperm motil, %sperma hidup, % sperma abnormal,dan rangking keaktifan sperma. Untuk mengetahui ada atautidaknya perbedaan yang bermakna antar perlakuan dalam kelompok dilakukan uji Anava satu arah,bila terdapat perbedaan bermakna dilanjutkan uji BNJ untuk membandingkan angka rata-rata antarkelompok perlakuan. Pemberian air rebusan akar Caesalpinea bonduc pada mencit menunjukanpengaruh perbedaan secara signifikan pada jumlah (%) sperma abnormal antara dosis 0,0 μl dengan24,0 dan 32,0 μl serta antara 16,0 μl dengan 24,0 dan 32,0 μl; Jumlah (%) sperma hidup terdapatperbedaan antara dosis 0,0 μl dengan 16,0 , 24,0 dan 32,0 μl; Jumlah (%) sperma motil terdapatperbedaan antara dosis 0,0 μl dengan 24,0 dan 32,0 μl; Rangking keaktifan sperma terdapatperbedaan antara dosis 0,0 μl dengan 24,0 dan 32,0 μl. Pemberian rebusan akar aru berpengaruhsecara signifikan terhadap peningkatan persen sperma abnormal, dan penuruan persen sperma hidup,motil dan keaktifan.AbstractIndonesia has many species of plants that have potent of medicine, one of them cold be as antraditional contraception medicine ussually used to limit child or sterilizatiom such as root boiledwater of Caesalpinia bonduc. The aims of this study is to know the effect of root boiled water ofCaesalpinia bonduc as traditional contraception medicine on quality of sperm maturation ofepididymis of mice (Mus musculus). Mice were choosed radomly and doses gouped: 0,0 (K0 ), 16,0(K1) 24,0 (K2) and 32,0 (K3) μl/gram body weight/day. Each of group was replicated 5 times.Treatment were given by oral with using gavage needle for ten days. Surgery was carried out on firstday after completing treatments. Examination of epidymis sperm by counting number of spermproduction, percentage of motil, live, abnormal sperm, and rank of sperm motility. In order to knowthe deferences between control and treated group, was used one way anava analysis and analysedvalue for comparing between teratment group. The treatments of root boiled water of Caesalpiniabonduc have effect significantly on percentage of abnormal sperm between dose of 0,0 μl with 24,0and 32,0 μl, also between 16,0 μl with 24,0 and 32,0 μl. Percentage of live sperem is different betweendose of 0,0 μl with 16,0 , 24,0 and 32,0 μl. Percentage of motile sperm is defferent significantlybetween dose of 0,0 μl with 24,0 and 32,0 μl. Meanwhile percentage of motile rank has differencebetween dose of 0,0 μl with 24,0 and 32,0 μl. The treatment of root boiled water of Caesalpiniabonduc has effect signifiantly on increasing percentange of sperm abnormality, decreasingpercentage of life sperm, motile and rank of motile of mice sperm.


1994 ◽  
Vol 193 (1) ◽  
pp. 79-95 ◽  
Author(s):  
J Claiborne ◽  
J Walton ◽  
D Compton-Mccullough

A number of studies have implied a linkage between acid­base and ion exchanges in both freshwater and seawater fish, although little is known about the branchial and renal acid­base transfers involved as the animals move between different salinities. To investigate the role of these transfers in a marine teleost fish as it is exposed to a dilute environment, we measured plasma acid­base values and net movements from fish to water of NH4+, HCO3- and H+ in long-horned sculpin (Myoxocephalus octodecimspinosus) placed in 100 %, 20 %, 8 % or 4 % sea water for 24­48 h. Renal excretion of H+ was also monitored in fish exposed to 4 % sea water. Sculpin proved to be somewhat euryhaline for they were able to maintain plasma ion and acid­base transfers in hypo-osmotic (20 %) sea water, but could not tolerate greater dilutions for more than several days. Plasma pH and carbon dioxide concentration (CCO2) increased in the 20 % and 8 % dilution groups, with CCO2 nearly doubling (control, 4.56 mmol l-1; 8 % group, 8.56 mmol l-1) as a result of a combined increase in the partial pressure of plasma CO2 (PCO2) and [HCO3-]. During a 44­46 h exposure, HCO3- transfers increased progressively in the most dilute water, with animals in the 8 % and 4 % groups exhibiting a net H+ loss that was smaller than that of seawater fish (control, 5.1 mmol kg-1; 8 %, 0.9 mmol kg-1; 4 %, -2.9 mmol kg-1). Animals exposed to 4 % sea water for 24 h and then returned to normal sea water had a variable plasma pH, an elevated CCO2 and a net efflux of H+ that effectively stopped (control, 0.10 mmol kg-1 h-1; 4 %, 0.02 mmol kg-1 h-1; seawater recovery, 0.20 mmol kg-1 h-1) during the low-salinity period. Renal acid excretion remained relatively constant throughout the experiment but only made up a significant portion (approximately 40 %) of the total acid transfers during the 4 % dilution period (control rate approximately 3 µmol kg-1 h-1: 3 % of branchial rate). We postulate that the increase in plasma CCO2 during exposure to low salinity may be due to mobilization of base from the intracellular bone compartment. The decrease in external salinity could induce base loss by alteration of gill ion exchanges (Na+/H+, Cl-/HCO3-) and/or changes in branchial HCO3- permeability. For the first time, we have shown that the effects of a dilute environment on acid­base transfers may be an important limitation to the survival of a euryhaline species in brackish or fresh water.


1948 ◽  
Vol 25 (4) ◽  
pp. 344-352
Author(s):  
LORD ROTHSCHILD

1. Echinus esculentus spermatozoa are normally motionless in undiluted semen. They become active when the semen is diluted with sea water or seminal plasma obtained by gentle centrifugation (1500 r.p.m., 12 cm. radius, for 15 min.). 2. A sperm-immobilizing substance can, however, be obtained in seminal plasma by more prolonged centrifugation of semen. 3. Spermatozoa can be made motile in undiluted semen by increasing the O2 tension in the atmosphere surrounding the semen. 4. This O2 activation is completely inhibited by N2; the N2 effect is reversible. 5. Measurements of seminal O2 tension were made with an O2 electrode. The O2 tension of semen is low, being at most 15 mm. Hg. It was not possible to decide whether this residual tension was due to O2 or some other substance reacting at the electrode. 6. The K content of seminal plasma is about 1·55 mg./ml., which is four times higher than that of perivisceral fluid or sea water. 7. The pH of semen is lower than that of sea water, being approx. 7·5. 8. Neither K concentration nor pH is responsible for the inactivity of sperm in semen. 9. A hormone, Androgamone I, is often considered to be responsible for the inactivity of spermatozoa in non-mammalian semen. No support has been found for this view and it is concluded that in E. esculentus semen the spermatozoa are motionless through lack of O2.


2006 ◽  
Vol 18 (2) ◽  
pp. 249
Author(s):  
G. Magarey ◽  
J. Herrick ◽  
K. Thiangtum ◽  
W. Tunwattana ◽  
W. Swanson

Wild populations of fishing cats (Prionailurus viverrinus) in Southeast Asia are in decline, primarily due to habitat loss. Because the fishing cat population in North American zoos is small (n = 69) and inbred (F = 0.17) with relatively low genetic variation (86%), infusion of new founder genes from Asia is a conservation priority. Importation of cryopreserved semen for use with IVF and ET may offer one alternative to the international transport of living animals. In this study, our objectives were to (1) compare motility longevity of fresh vs. frozen-thawed fishing cat spermatozoa in two culture media, (2) evaluate ovarian responses to exogenous gonadotropins, and (3) assess development of IVF embryos produced with fresh vs. frozen-thawed spermatozoa. Raw semen was collected via electroejaculation from male fishing cats (n = 4), divided into groups, and washed. Two sperm pellets were resuspended in either Ham's F10 medium (HF10; with 5% FBS) or our feline optimized culture medium (FOCM; with 0.4% BSA); another pellet was diluted in TEST egg yolk, cooled to 5�C over 3 h, glycerated (4%), and cryopreserved in straws over LN2 vapor. Frozen sperm samples were thawed, washed, and diluted in either HF10 or FOCM. Fresh and frozen-thawed sperm motility (percent motile, rate of forward progress) in each medium (10 � 106 motile sperm/mL) was assessed (at 0, 1, 3, and 6 h) in microdrops under oil during culture (38�C; 6% CO2 in air). Female fishing cats (n = 10) were treated with exogenous gonadotropins (150 IU eCG, 100 IU hCG, 85-h interval) and ovarian follicles were aspirated laparoscopically. Recovered oocytes were inseminated with fresh (2 � 105 motile sperm/mL) or frozen-thawed (5 � 105 motile sperm/mL) spermatozoa in FOCM microdrops; resulting embryos were either cryopreserved or cultured in FOCM (with 5% FBS added at 72 h post-insemination) for 7 days. Sperm motility over time did not differ (P > 0.05) between media for either fresh or frozen-thawed samples; however, across media, frozen-thawed sperm motility was lower (P < 0.05) and declined faster (P < 0.05) compared to fresh spermatozoa. Females produced an average (�SEM) of 9.8 � 2.9 mature ovarian follicles, allowing recovery of 7.3 � 2.6 high-quality oocytes per female. Oocyte cleavage percentage at 42 h p.i. was lower (P < 0.05) with frozen-thawed spermatozoa (38%, 11/29) compared to freshly collected spermatozoa (68%, 17/25). Overall, 35% (6/17) of cultured embryos developed to blastocysts with no difference (P > 0.05) between embryos produced with frozen-thawed (4/11) vs. fresh (2/6) spermatozoa. Although fishing cat sperm motility and fertility appear compromised after cryopreservation, our results demonstrate the ability of frozen-thawed spermatozoa to produce IVF embryos that are capable of developing to blastocyst stage in vitro. This work was supported by (NIH RR015388).


2020 ◽  
Vol 32 (2) ◽  
pp. 183
Author(s):  
Y. Honkawa ◽  
T. Fujikawa ◽  
N. Miura ◽  
C. Kubota

It is difficult to maintain sperm in liquid storage for a long time, compared with permanent frozen storage in liquid nitrogen. Antioxidants have been reported to improve the quality and fertility of liquid-stored semen. In this study, we investigated whether antioxidants can extend the motility and fertility of frozen-thawed sperm in liquid storage. Frozen-thawed semen from one Japanese black bull (one ejaculate) was diluted in Tris-citrate-fructose (TCF) diluent with 10% (v/v) egg yolk to a sperm concentration of 1×107 spermmL−1. The antioxidants β-mercaptoethanol (βMe) and glutathione (GSH) were added independently, at various concentrations (0.1, 0.5, 1, and 5mM) to sperm suspensions, and these preparations were compared with Control (no added antioxidant). Sperm suspensions were packaged in centrifuge tubes and placed at 17°C in air and monitored daily until sperm motility had stopped (up to 14 days). Sperm motility was analysed by the Sperm Motility Analysis System (SMAS; Ditect Co. Ltd), and the percentage of progressively motile sperm (straight-line velocity (VSL) of &gt;25μm s−1; Grade A classified by WHO manual), compared with that recorded on Day 0 (100%), was determined each day. For evaluation of fertilizing ability, after incubation in liquid storage for 0, 3, 5, and 7 days, sperm were used for IVF with invitro-matured oocytes (30 oocytes per treatment, three replicates). Embryo development was recorded as the proportion of embryos that reached blastocyst by 8 days after IVF. Data for motility were analysed using one-way ANOVA with Tukey test, and embryo development using chi-squared test. A P-value&lt;0.05 was considered statistically significant. At 7 days, the percentage of progressively motile sperm was significantly higher for 0.5, 1, and 5mM βMe than for Control (30.8%, 48.1%, and 50.3%, vs. 0%, respectively). Treatments with 1 and 5mM βMe maintained some sperm progressive motility for 14 days (9.5% and 14.5%). Treatment with GSH showed the same trend at 7 days (32.2%, 36.3%, and 13.7% for 0.5, 1, and 5mM, vs. 0% for Control); 1 and 5mM GSH maintained sperm progressive motility over 10 days (24.8% and 4.4%). In both antioxidant treatments, embryo development was achieved with sperm stored for up to 5 days (Day 0 vs. Day 5 for 0.1mM βMe: 17.6% vs. 13.8%; for 1.0mM GSH: 26.0% vs. 6.7%; for Control: 17.6% vs. 0%). In this study, antioxidants extended both motility and fertility of frozen-thawed bovine sperm in liquid storage. This result suggests the possibility of application to AI using liquid-stored bovine semen.


1977 ◽  
Vol 70 (1) ◽  
pp. 213-220
Author(s):  
DAVID H. EVANS

1. Four species of marine teleosts were shown to possess an external-NH4-inhibited Na uptake from 1 mM-NaCl solutions. The inhibition was not due to changes in the transepithelial potential. 2. Injection of 2 μM-NH4/g fish stimulated Na uptake by Opsanus beta and also stimulated ammonia efflux, 50% of which was dependent upon external Na. 3. The ammonia efflux from three species was partially dependent upon external Na. 4. Na/NH4 exchange in O. beta could be reversed so that 22Na efflux could be stimulated by the addition of 200 mM-NH4 to the external solution. 5. These studies show clearly that marine teleosts possess an Na/NH4 exchange system in sea water which results in a net influx of Na into the fish.


1976 ◽  
Vol 65 (3) ◽  
pp. 529-538
Author(s):  
J. C. Carrier ◽  
D. H. Evans

(1) The marine teleost fish, Lagodon rhomboides, can only tolerate fresh water (5 mM Na) if Ca is also present (10 mM). Transfer to Ca-free fresh water is followed by a substantial increase in radioactive Na efflux with little or no change in the transepithelial potential. Addition of the chelating agent EDTA (2 mM) further increases Na efflux. Fish left in Ca-free fresh water for 2-5 h die with a total body Na less than 50% of that found in animals acclimated to Ca-supplemented fresh water. (2) Rates of Na uptake were measured on either sea-water-acclimated or Ca-supplemented fresh water-acclimated fish transferred to various low Na media. In both cases Na uptake has a high Km, is saturable, inhibited by external NH4, H and amiloride, and is not related to changes in the trans-epithelial potential. (3) It is suggested that L. rhomboides is dependent upon external Ca to decrease diffusional Na loss in low salinities so that a relatively inefficient Na uptake can balance diffusional and urinary Na loss.


2016 ◽  
Vol 50 (1) ◽  
Author(s):  
J. B. Patel ◽  
A. J. Dhami

Sixty semen ejaculates from 10 mature bulls, 5 each of Jafarabadi and Mehsana breed, were studied for sperm motility and velocity parameters of fresh and frozen-thawed spermatozoa using computer assisted sperm analyzer (CASA). The mean values of motile and progressively motile spermatozoa observed in fresh semen of Jafarabadi and Mehsana bulls (79.77±1.62 and 61.80±1.85, and 78.90±1.22 and 61.37±1.58%) were highly significantly (P<0.01) reduced (51.20±1.57 and 33.20±1.45, and 52.10±1.70 and 34.30±1.54 %, respectively) in post-thawed semen. The average path velocity, straight line velocity and curvilinear velocity (μm/sec) of spermatozoa of Jafarabadi and Mehsana bulls noted in fresh semen were also reduced highly significantly (P<0.01) in frozen-thawed semen. Among the other velocity parameters, amplitude of lateral head displacement (μm), elongation (%) and medium motile sperm (%) increased, while beat-cross frequency (Hz), straightness (%), linearity (%), sperm area (μm<sup>2</sup>) and rapidly motile sperm (%) decreased significantly in post-thawed sperms when compared with the fresh sperm of both Jafarabadi and Mehsana bulls. The initial motility and live sperm per cent were significantly correlated with CASA traits of fresh and frozen-thawed semen, and all the sperm motility and velocity traits of fresh and frozen-thawed semen assessed by CASA were significantly interrelated among both the breeds. The interrelationships were stronger in Mehsana bulls as compared to Jafarabadi bulls.


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