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Effect of Diluents on the Cold Storage of Sperm in Sea Urchin, Hemicentrotus pulcherrimus
고강희,강경호,김재민 한국발생생물학회 2003 발생과 생식 Vol.7 No.1
말똥성게 정자의 냉장보존을 위한 여러 가지 희석액의 보존 효과를 비교하였다. 다양한 농도의 포도당 용액, 인공해수(ASW)를 희석액으로 하여 정자를 4에서 보존하였다 보존효과는 정자 활성지수 (SAI), 생존율 및 말똥성 게 난에 대 한 수정률로 평가하였다. ASW와 1.2 M glucose액을 희석 액으로 하여 30일간 냉장보존하였을 때 가장 높은 운동성 및 생존율을 유지하였다. 정자의 냉장보존을 위한 희석액의 적정 pH는 7.0∼8.0 이었다. 정자 A series of experiments were conducted to compare the effects of various diluents in cold storage on the sea urchin, Hemicentrotus pulcherimus sperm. Various diluents of glucose solutions, artificial sea water(ASW) and 50% ASW were used to store the sperm at 4. The storage effect was evaluated using sperm activity index(SAI), survival rate of sperm and fertilization rate to egg. ASW and 1.2 M glucose were found to be better diluents which maintained high motility and survival rate of sperm f3r a storage period of 30 days. Optimal pH of diluent to store the sperm at 4 is 7.0∼8.0. In order to keep high SAI and survival rate of sperm, addition of 400 ppm neomycin into the diluent revealed the best storage results.
Mechanisms for the Initiation of Sperm Motility
고강희,강경호,장영진 한국발생생물학회 2003 발생과 생식 Vol.7 No.2
정자의 운동개시는 수정시에 정자와 난자가 만나기 위한 전제조건이다. 동물의 정자는 CAMP와Ca2'의 조절기구에 의해서 정자의 운동개시가 일어난다. 정자운동 활성 및 개시를 위한 세포 신호전달기구는 멍게류와 연어과 어류에서 많은 연구가 이루어져 왔다. 멍게류의 경우, 난에서 분비되는 정자 활성 및 유인물질(sperm-activating and -attracting factor)은 정자 활성 및 난으로의 유인을 위하여 외부의 을 요구한다. 한편 연어과 어 Initiation and activation of sperm motility are prerequisite processes for the contact and fusion of male and female gametes at fertilization. The phenomena are under the regulation of CAMP and in vertebrates and invertebrates. Mammalian sperm requires and cyclic AMP for the activation of sperm motility. Cell signaling for the initiation and activation of sperm motility has been well studied in the ascidians, Ciona intestinalis and C. savignyi and salmonid fishes. In Ciona, whose cell signaling for activation of sperm motility has been established, the sperm-activating and -attracting factor released from unfertilized egg requires extracellular for activating sperm motility and eliciting chemotactic behavior of the activated sperm toward the egg. On the other hand, the cyclic AMP-dependent phosphorylation of protein is essential for the initiation of sperm motility in salmonid fishes. A decrease in the environmental Ti concentration surrounding the spawned sperm causes a li efflux and influx through the specific channel and dihydropyridine-sensitive L-/T- type channel, respectively, thereby leading to the membrane hyperpolarization and influx. The membrane hyperpolarization synthesizes cyclic AMP, which triggers the luther Process of cell signaling, i.e., cyclic AMP-dependent protein phosphorylation, to initiate sperm motility in salmond fishes.almond fishes.
무지개송어(Oncorhyndhus mykiss)정자의 동결보존
고강희 전남대학교 수산과학연구소 2014 수산과학연구소논문집 Vol.22 No.-
This experiments was performed to identify the suitable cryoprotectants, freezing rate and, thawing rate for sperm cryopreservation in the rainbow trout, Oncorhynchus mykiss. In cry-opreservation experiments, 300 mM glucose solution containing 20 mM KCl, 20 mM Hepes and, pH 7.6 was used as a diluent, and 10% dimethyl sulfoxide (DMSO) was used as a cryoprotectant, and 10 % egg yolk was used as an addictive material. As a results of cry-opeservation, when 10 % DMSO was used as a cryoprotectant, sperm motility showed the highest rate at the height of 2 ㎝ away from the liquid nitrogen. Thawing at 10 ℃ for 30 seconds displayed the best result in the thawing rate.
고강희,김정우,김수철,최명락,한경호,이원교,최갑성 한국응용생명화학회 2015 Applied Biological Chemistry (Appl Biol Chem) Vol.58 No.5
Carbonic anhydrases (CAs) play a key role in physiological functions such as pH homeostasis, calcification, photosynthesis, and ionic regulations. Among CA isozymes, CA I is a specific marker for the cytoplasm or apical cell membranes of colonic epithelial cells, and it is related to enterocyte proliferation in mammals. The major aim of the present study was to investigate the regional, cellular, and subcellular localization of CA I in a fish, rainbow trout, Oncorhynchus mykiss. The CA I was identified in the gills, heart, and intestine of the rainbow trout. CA I protein detected in the tissues of the rainbow trout was similar to a CA III with a molecular weight of 29 kDa. Immunohistochemical results demonstrated that CA I was localized in the various cells of gills, heart, muscle, and intestine of the rainbow trout.
Effects of Osmolality and $Ca^{2+}$ on Sperm Motility in Marbled Sole, Limanda yokohamae
고강희,장영진,임한규,KHO Kang Hee,CHANG Young Jin,LIM Han Kyu The Korean Society of Fisheries and Aquatic Scienc 1997 한국수산과학회지 Vol.30 No.5
문치가자미의 성숙한 수컷 15마리를 재료로 하여 삼투질농도와 $Ca^{2+}$이 정자의 운동성에 미치는 영향을 파악하였다. 문치가자미 정자는 전해질 및 비전해질 용액에서 정장의 삼투질농도인 $351{\pm}23.4mOsm/kg$ 이하의 저장액에서 운동이 억제되었으나, 삼투질농도 $466\~l,185mOsm/kg$에서는 활발한 운동을 보였으며, 1,698 mOsm/kg 이상에서 다시 운동이 중지되었다. 인공해수를 희석액으로 하여 pH를 달리하였을 때, 정자는 $pH\;5\~9$에서 운동지수 $3.75\~4.00$로 운동성이 활발하였으나, pH 4에서는 운동지수 2.50로 미약하였다. $Ca^{2+}$의 특이 chelator인 EGTA가 포함된 $Ca^{2+}$ 결여 인공해수 (tris-HCl, pH 7.4)에 정액을 희석하였을 때, EGTA의 농도 증가에 따라 정자 운동성이 낮아졌으며, 3mM 이상의 농도에서는 완전히 정지되었다. 운동이 억제된 정자에 $Ca^{2+}$을 서서히 첨가한 결과, $Ca^{2+}$ 농도 0.25mM 이상부터 운동이 재개되었다. 또한 인공해수속의 정자는 calmodulin 억제제인 TFP의 농도의 증가에 따라 운동성이 감소하여 0.5mM에서 완전히 정지하였다. A series of experiments were conducted to study effects of osmolality and $Ca^{2+}$ on sperm motility in marbled sole, Limanda yokohamae. Spermatozoa were immotile when the milt was mixed with solutions (electrolyte or non-electrolyte) of lower osmolality than the average seminal plasma osmolality (351 mOsm/kg), but became motile after mixing milt with an hyperosmotic solution. However, with the osmolality above 1,639 mOsm/kg spermatozoa ceased their movement. When the milt was suspended in the $Ca^{2+}$ free artificial seawater (ASW) containing ethylenglycoltriamine (EGTA), the percentage of motile spermatozoa decreased in proportion to the increase of EGTA. Most spermatozoa were quiescent in a medium containing 3 mM EGTA. The motility of spermatozoa prevented by 3 mM EGTA was recovered by the subsequent addition of $Ca^{2+}$ over the concentration of 0.25 mM. Effects of calmodulin antagonist, trifluoperazin (TFP) on spermatozoa were examined to elucidate the possible functions of calmodulin in sperm motility. TFP immobilized spermatozoa 5n ASW at the concentration of 0.5 M. These findings suggest that calcium is an effective external factor in the initiation process of sperm motility.