• International Journal of Industrial Entomology and Biomaterials
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• v.16 no.1
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• pp.1-5
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• 2008

The present study was carried out with the aim to evaluate the viability of layings (delivery of fertilized and unfertilized eggs) with egg laying duration of elite bivoltine races. The temporal aspect of mating in terms of egg layings duration may also have impact on the number of eggs laid, pattern of egg laying and their viability. After different interval of egg laying duration, moth of Bombyx mori are removed from oviposition site after they have completed egg laying. Present investigation confirmed that there are no significant difference in relation to the occurrence of viable (fertilized) and non viable (unfertilized) eggs and duration of oviposition. Percentage of unfertilized eggs varied from 2.41 to 3.42% in case of CSR-3, 2.59 to 3.62% in case of CSR-6, 2.82 to 3.66% in case of CSR-16 and 2.58 to 3.40% in case of CSR-17 in different treatments. The occurrence of unfertilized eggs are also not significant in the different treatments specially in those eggs which were laid 24 hours after oviposition and kept for 4 months hibernation schedule.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.1
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• pp.1-7
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• 2009

Quality of seed in the mulberry silkworm, Bombyx mori (L.) is determined by many important factors, wherein unfertilized eggs play an important role. Unfertilization of eggs are caused by several reasons such as, abnormality in the sexual organs of the male and female, abnormal development of the micropylar end of the egg, unfavorable environmental conditions during spinning, cocoon preservation, imperfect handling of moths, mating, ovipostion, cold storing of pupae / moths and indiscriminate use of male moths etc. Though the presence of unfertilized eggs would in no way affect the fertilized ones and their quality directly, the frequency of their occurrence underrates the quality and brings down the hatching percentage. Lower the occurrence of unfertilized eggs, higher is the rating of seed quality. Of the various intrinsic and extrinsic factors and events involved in egg deposition of an adult silk moth, mating is an instinct and a biological obligation for the ultimate perpetuation of the species and a must to provide stimulus for oogenesis and bring about biochemical changes in the spermatophore of the silkworm in order to ensure the presence of sufficient number of normal sperms and testicular fluid in the female reproductive organ, activating ovulation and accelerating oviposition behavior and egg deposition. An attempt has been made in this article to briefly elucidate the characteristics of unfertilized eggs, causes of their occurrence and its impact as well as the significance in silkworm seed production.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.245-251
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• 1989

In order to manifest the presence of Na-K pump and its property on the unfertilized egg membranes of mouse, membrane potential was recorded under the physiological condition (at $37^{\circ}C$ and 4mM $Ca^{2+}$). After an induction of superovulation, the fresh eggs with zona pellucida were collected from mouse oviduct. Transient hyperpolarization as pump action was recorded after the switch into the high potassium perfusate (15mM $K^+$) from K-free perfusate, and the difference between membrane potential observed just before the perfusion of high potassium solution and the maximal membrane potenlial during the perfusion of high potassium solution was regard as pump activities. The results observed were as follows, 1. Resting mombrane potential was depolarized under the treatment of $10^{-5}M$ ouabain. 2. Pump activities of the unfertilized mouse eggs were $-3.38{\pm}0.61mV$ ($Mean{\pm}SD$, n=6), recorded as transient hyperpolarization due to the electrogenic property. 3. Pump activities were blocked by both treatment of $10^{-5}M$ ouabain and perfusion of Nafree solution, while increased by high $Na^+$ (300mM) perfusion ($-7.45{\pm}0.75mV$, n =2). 4. Hyperpolarization due to pump activity was not altered by $Mn^{2+}$. 5. Above results confirm the presence of ouabain-sensitive Na-K pump, which affected the membrane potential directly, on the unfertilized egg membranes of mouse.

• The Korean Journal of Physiology
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• v.23 no.1
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• pp.35-42
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• 1989

The present study was performed to observe the effects of cations on resting membrane potential and pump activity in the unfertilized eggs of ICR strain mice. After an induction of superovulation, the fresh eggs with zona pellucida were collected and the membrane potentials were recorded. Recordings of membrane potential in this study was obtained from the physiological conditions ($37^{\circ}C$ and 4mM Ca in standard solution), differently from the another reports with unphysiological conditions (room temprature and high Ca in standard solution) for a stable and long-lasting observations. Presented data was obtained within 6 hours after collection from the oviduct. The results observed are as follows, 1) Resting potential of the unfertilized eggs was $-25.8{\pm}3.8mV$ $(Mean{\pm}Se,\;n=31)$. 2) As the K ion concentration was increased, resting membrane potential was depolarized but showed hyperpolarization with $K^{+}$ below 25mM. 3) Alteration of the resting membrane potential for the changes of $Na^{+}$ concentration were hardly observed, while resting potential was hyperpolarized as $Ca^{2+}$ concentration was increased. 4) Pump activity as transient or prolonged hyperpolarization was $-2.29{\pm}0.75mV$ $(Mean{\pm}Se,\;n=16)$, the hyperpolarization was increased in both amplitude and duration under the 10mM $Ca^{2+}$ solution. 5) Hyperpolarization due to pump activity was decreased or disappeared by $5{\times}10^{-5}\;M$ ouabain treatment and could not be observed under the both Na-free and Ca-free solutions. 6) Above results are likely to suggest that the resting potential of the mouse unfertilized eggs is affected to mainly by Ca-dependent K conductance and Na-Ca exchange mechanism and that there is pump activity coupling between $K{+}$, $Na^{+}$ and $Ca^{2+}$.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.117-121
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• 2004

The patterns of diapause-associated proteins of silkworm eggs were analyzed by two-dimensional (2-D) gel electrophoresis. Among the hundreds of spots on the 2-D gels, at least two proteins were considered to be associated with diapause. A protein, spot 4, with an approximate molecular weight of 38 kDa and pI 6.1 was observed in the HCI-treated, cold-treated, and diapause eggs, respectively. Spot 4 was undetectable in unfertilized eggs and non-diapause eggs at two days after oviposition, suggesting that this protein may be associated with the entrance to diapause. A protein, spot 11, with an approximate molecular weight of 21 kDa and pI of 61 was detected in the unfertilized, HCl-treated, and cold-treated eggs, respectively, after oviposition by normal moths. In diapausing eggs, a protein corresponding to spot 11 was observed in 3-, 5-, and 30-day-old eggs, while the protein was not detected one-day-old eggs. The protein corresponding to spot 11 was not detected in unfertilized and non-diapause eggs obtained from subesophabeal ganglion (SG)-extirpated moths either. Spot 11 was also considered to be a diapause specific protein, which occurred at only early embryonic stage under the control of diapause-downregulated gene.

• Korean Journal of Animal Reproduction
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• v.13 no.3
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• pp.188-194
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• 1989

These experiments were carried out to investigate fertilizable and developmental ability after zona drilling the unfertilized eggs and the eggs not fertilized by the 1st insemination. The results of in vitro fertilization of the mouse eggs treated by using micromanipulation and acid tyrode's solution with capacitated epididymal spermatozoa were as follows. In the case of ovulated unfertilized eggs, according to sperm count(106, 105, 104 and 103/ml) the rates of in vitro fertlilization treated by zona drilling were 86.0%, 82.0%, 70.0% and 54.0%, respectively, and those of control were 58.0%, 52.0%, 12.0% and 8.0%, respectively. The rates of in vitro fertilization of zona drilled eggs were significantly high compared with those of control, and there were no significant difference between two groups. According to the sperm count the zona drilled eggs developed to the blastocysts were 51.4%, 40.5%, 23.3% and 17.4% and those of control were 35.7%, 26.3%, 0% and 0%, respectively. Also, in the eggs not fertilized by 1st insemination, the fertilization rates of oocytes reinseminated after zona drilling was significantly higher(83.5%) than that of control(34.7%), and the rates of polyspermy were similar. The rates of development to the blastocysts was 18.6% in the zona drilling treated eggs, and that of control was 27.3%, there was no significant difference between two groups. These results indicated that oocytes not fertilized by 1st insemination as well as ovulated unfertilized eggs could be fertilized, improve fertilizing rates by zona drilling treatment, and development potential were normal.

• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.157-164
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• 1992

Changes in the both inward current and conductance of membrane by the fertilization were observed using the one microelectrode voltage clamp(or switch clamp) technique. Unfertilized eggs and both 1- and 2-cell stage eggs after fertilization were donated from the superovulated mouse (ICR, more than 6 weeks old) treated with PMSG(pregnant mare serum gonadotropin, Sigma) and HCG(human chorionic gonadotropin, Sigma) and naturally mated ones, respectively in this experiment. Membrane potential was held at -90mV and the voltage step was applied from -80mV to 50mV with interval of 10mV or 20mV for 300ms. since both of amplitudes and time courses in the membrane currents were various according to the states of cells and clamping condition, results were presented by their $averages{\pm}SEM$(standard mean error)and ratios or percentages. Inward currents began to appear in response to the step depolarization from -60mV and reached its maximum at -50mV. However, since the potential was not clamped evenly during the voltage step, current-voltage(I-V) relationship might be positively shifted 10 or 20mV. From the steady-state currents plotted in the I-V curve, outward rectification was markedly observed. Peak inward currents$(i_{in})$ at -50mV were $-0.62{\pm}0.23nA$(n=4),$-0.52{\pm}0.25nA$(n=5) and $-0.37{\pm}0.25nA$(n=6), in the 1-cell stage, 2-cell stage fertilized eggs and in the unfertilized eggs, respectively. Pure inward current (difference between steady-state and peak, $i_{in. pure}$) were $-1.01{\pm}0.23nA$, $-0.69{\pm}0.43nA$ and $-0.68{\pm}0.29nA$, respectively in the 1-cell stage fertilized eggs, unfertilized eggs and 2-cell stage fertilized eggs. These results suggested that the outward current in fertilized eggs of 2-cell stage was more increased than those in the unfertilized eggs. Pure inward currents in the all stages of eggs showed a similar fashion in the I-V relationship from -50mV to 50mV and reversal potential at 50mV. Time constant of inactivation$({\tau})$ in the inward current was decreased as the membrane potential was depolarized in the unfertilized and 2-cell stage eggs but in the 1-cell stage eggs t was not likely to be affected significantly. Slope conductances were 14.2nS, 8.9n5 and 7.7nS in the 1-cell, 2-cell stage fertilized eggs and the unfertilized eggs, respectively. Membranes between two cells within a zona pellucida seem to be electrical-connected in the 2-cell stage eggs from the observation made in the analysis for the electronic spread and decay to the current stimuli. Both of inward current and membrane conductance were increased after fertilization in the mouse eggs. Inward current seems to be carried by the same ion or through the same channels up to the 2-cell stage and ion that carried inward current was thought to play important function after fertilization in the mouse eggs.

• Journal of Sericultural and Entomological Science
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• v.39 no.1
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• pp.17-21
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• 1997

Oral application of fenoxycarb, O-ethyl N-(2-(4-phenoxyphenoxy)ethyl) carbamate, the commercial formulation Insegar, on the 1st day of the 3rd instar made no difference in the egg laying and the hatchability as compared to 750 grains for the number of eggs laid, 53 grains for the number of the resideual eggs unlaid, 749 grains for the number of fertirized eggs, 97% for the total hatchability, and 95% for the useful hatchability as in the control but on the first day of the 4th instar, the number of the remainder eggs in the ovary increased by 2.4 folds, and reduced number of the eggs laid, number of unfertilized eggs, the hatchabillity and useful hatchability by 39%, by 66%, by 68% and by 72%, respectively. When diluted Insegar was applied topically every other two days from the pupae to the moth at concentration of 10-6, 10-7, 10-8, 10-9, 10-10 the number of eggs laid, the number of the unfertilized eggs, the total hatchability and the useful hatchability were decreased. More severe decrease took place as concentration lowered and the earlier application.

• The Korean Journal of Physiology
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• v.27 no.1
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• pp.93-105
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• 1993

The present study was performed to investigate the properties of ionic currents elicited by voltage pulses in the unfertilized eggs of mouse and hamster by using the whole cell voltage clamp techniques and to find out if there are any differences in properties between eggs of the two rodents. In addition, the modulatory effect of G proteins and protein kinase C (PKC) on the ionic channels were observed. The inward current in hamster eggs was shown to be due to $Ca^{2+}\;current\;(i_{ca})$). The current voltage relations of these currents in hamster egg were analogous to those in mouse eggs. The amplitude of $i_{ca}$ in the hamster egg was larger than that in the mouse egg ($-3.12{\pm}1.07\;nA\;vs.\;-1.71{\pm}0.71\;nA,\;mean{\pm}\;SD$). These results suggest that the $Ca^{2+}$ channels in both kinds of eggs have similar channel properties but their density, and/or conduct ance per unit area is higher in hamster eggs than in mouse eggs. Outward currents in eggs of both mouse and hamster were carried by $K^+$. In hamster eggs, they appeared to comprise at least two components; a transient outward component ($i_{to}$) and a steady state component ($i_{\infty}.$ The $i_{to}$ was found to be dependent on intracellular $Ca^{2+}$ concentration; whereas on the other hand $i_{\infty}\;was\;Ca^{2+}$-independent. $Ca^{2+}$ currents were increased in eggs treated with GTP (or $GTP{\gamma}S$) or fluoroaluminate ($AIF_4^-$). In the hamster egg these increments were antagonized by GDP (or $GDP{\beta}S$) application. In contrast to the enhancement of $i_{ca},\;i_k$ was reduced following GTP (or $GTP{\gamma}S$) perfusion in mouse eggs. The transient component ($i_{to}$) in hamster eggs was increased by adding GTP but decreased by phorbol ester, TPA or dioctanoyl glycerol (DOG). Simultaneous application of $GTP{\gamma}S$ and DOG suppressed $i_{to}$ more effectively than a single application or DOG or TPA. From the above results, we have shown that ionic currents elicited by voltage pulses existed in the unfertilized eggs of mouse and hamster. There are at least two types of currents, $i_{ca}\;and\;i_k$ in mouse eggs, while three types, $i_{ca},\;Ca^{2+}$-dependent $i_k$ and $Ca^{2+}$-independent $i_k$ exist in hamster eggs. ionic channels in these eggs may be regulated either directly by GTP and PKC or indirectly by the substances linked with GTP and PKC.

• Korean Journal of Veterinary Research
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• v.31 no.1
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• pp.33-40
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• 1991

For the observations of both the membrane properties and the excitability on the unfertilized eggs of female mice, changes of the membrane resistance and the membrane potential by hyerpolarizing current stimulation were recorded. As current-voltage relation was linear over the entire range (-180mV~+60mV), membrane resistance($R_m$) was calculated from the amplitude of electrotonic potential to a given stimulus current. Also the presence of anode-break excitation was confirmed. The results were as follows; 1. There was a linear relation between the membrane resistance and resting membrane potential, the expected input resistance was 61. 4M$\Omega$(resting membrane potential was $-18.9{\pm}8.7mV$, mean${\pm}$SD, n=30). 2. Transient depolarization with overshoot was generated just after hyperpolarizing current stimulus and showed the dependency of stimulus duration. 3. Transient depolarization lasted over 30ms, amplitude of these depolarization was increased by high $Ca^{{+}{+}}$(20mM) and inhibited by $Ca^{{+}{+}}$-antagonist, $Mn^{{+}{+}}$. 4. From the above results, it was suggested that the unfertilized mouse egg showed the characteristics of the excitable cell.