본 연구는 동자개 정자의 동결보존을 위해 동결보존제의 최적 농도와 적정 희석액을 구명하여 정자를 최상의 상태로 보존하여 인공종자를 생산하는데 목적이 있다. 실험은 3종의 희석액(Ⅰ: 300 mM glycose, Ⅱ: Kurokura extender, Ⅲ: Li extender), 4종의 동결보존제(dimethyl sulfoxide, ethylene glycol, methanol and glycerol)와 4개의 동결보존제 농도(5, 10, 15, 20%)를 조합하여 대하여 조사하였다. 동결보존한 정자를 해동한 후 정자의 생존율과 정자활성지수로 동결보존 제의 효과를 평가하였다. 희석액 Ⅲ(Li extender)과 10과 15%의 methanol을 조합했을 때 동자 개 정자의 생존율과 정자활성지수는 각각 66.9 ± 8.7, 67.3 ± 13.1%과 2.6 ± 0.4, 2.6 ± 0.5로 다른 희석액과 동결보존제보다 높았다.
The histological changes in the Ussurian bullhead, Leiocassis ussuriensis, and the Korean bullhead, Pseudobagrus fulvidraco, were observed during the early period of growth. The retinas size of both species increased in the 9 days post-hatching (DPH) (P<0.05). In the just-hatched Ussurian bullhead, the retina already consisted of six layers: the epithelial layer, ganglion cell layer, inner nuclear layer, inner plexiform layer, outer limiting membrane layer, and rod and cone layer. The Korean bullhead had the same components. At 50 DPH, the thickness of the retina was 538.0±7.19 μm in the Ussurian bullhead and 558.9±9.44 μm in the Korean bullhead. The relative thickness of each layer of the retina did not differ significantly in the two species. Although the growth of the Korean bullhead’s retina was faster, the relative thickness of each layer in the retina did not change during early development. After hatching, some parts of the tissue gradually became denser. Immediately after hatching, the kidney and midgut epithelium of the Ussurian bullhead and Korean bullhead were already formed and grew gradually thereafter. From 0 DPH to 30 DPH, the nuclear height in the midgut epithelium did not differ significantly between the two species, but at 50 DPH, it was 11.4±2.45 μm in the Korean bullhead and 9.9±2.13 μm in the Ussurian bullhead. During the experimental period, the major axes, minor axes, surface areas, and volumes of the proximal tubule cells in the kidney did not differ significantly between the two species. Thus, the early histological development of the Ussurian bullhead is similar to that of the Korean bullhead.
The histological changes in the Ussurian bullhead, Leiocassis ussuriensis, and the Korean bullhead, Pseudobagrus fulvidraco, were observed during the early period of growth. The retinas size of both species increased in the 9 days post-hatching (DPH) (p<0.05). In the just-hatched Ussurian bullhead, the retina already consisted of six layers: the epithelial layer, ganglion cell layer, inner nuclear layer, inner plexiform layer, outer limiting membrane layer, and rod and cone layer. The Korean bullhead had the same components. At 50 DPH, the thickness of the retina was 538.0±7.19 μm in the Ussurian bullhead and 558.9±9.44 μm in the Korean bullhead. The relative thickness of each layer of the retina did not differ significantly in the two species. Although the growth of the Korean bullhead’s retina was faster, the relative thickness of each layer in the retina did not change during early development. After hatching, some parts of the tissue gradually became denser. Immediately after hatching, the kidney and midgut epithelium of the Ussurian bullhead and Korean bullhead were already formed and grew gradually thereafter. From 0 DPH to 30 DPH, the nuclear height in the midgut epithelium did not differ significantly between the two species, but at 50 DPH, it was 11.4±2.45 μm in the Korean bullhead and 9.9± 2.13 μm in the Ussurian bullhead. During the experimental period, the major axes, minor axes, surface areas, and volumes of the proximal tubule cells in the kidney did not differ significantly between the two species. Thus, the early histological development of the Ussurian bullhead is similar to that of the Korean bullhead.
Morphometric changes in the Ussurian bullhead, Leiocassis ussuriensis, and the Korean bullhead, Pseudobagrus fulvidraco, were observed during the early period of growth. Yolk length, yolk height, and yolk volume in the two species decreased within 9 days post-hatching (DPH) (p<0.05). The body lengths and body heights of both species increased gradually to 150 and 130 DPH, respectively (p<0.05). The horizontal distance between the anteriormost extension of the head and the anterior insertion of the pectoral fin, the anteriormost extension of the head × the verticality position of the anterior insertion of the primary dorsal fin rays, and the anterior insertion of the primary dorsal fin × the anterior insertion of the pectoral fin were greater in the Korean bullhead than in the Ussurian bullhead (p<0.05). However, the relative sizes of the head region, pectoral fin, ventral fin, and anal fin were greater in the Ussurian bullhead than in the Korean bullhead (p<0.05), and relative body depth and the size of the outer-mandible barbel were greater in the Korean bullhead than in the Ussurian bullhead (p<0.05). The growth curves of the morphometric characteristics of both species were divided into three types.
Korean bullhead (Pseudobagrus fulvidraco) was collected from the Kum River areas of Kangkyung-eup, Nonsan city, Chungcheongnam-do, Korea, from April to June, 2012 and was fertilized in order to observe egg development and temperature-related cleavage rates and mitotic intervals (). The fertilized eggs were separative, demersal and light yellowish with in diameter, and did not contain oil globules. The first cleavage stages were 90 min, 80 min, 60 min and 50 min at , , and , respectively. At higher temperatures, eggs developed faster and underwent further identical development. For Korean bullhead, were min at , min at , min at and min at . There were strong negative correlations between the and water temperatures at all points studied (Y=-1.13X+58.15, =0.98, n=30, where Y is and X is temperature). The results obtained in this work will be helpful for chromosome manipulation by use of cleavage frequency data and data in Korean bullhead.
눈동자개, Pseudobagrus koreanus의 성분화 과정을 밝히기 위해 부화 직후부터 250일령까지 생식소의 분화 및 발달을 조사하였다. 원시생식세포는 부화 후 1일(전장: 6.63~6.95 ㎜)에 나타났으며, 부화 후 5일(7.50~9.36 ㎜)에 생식융기를 형성하는 원시생식소 구조를 나타내었다. 부화 후 25일(11.58~13.21 ㎜)에서는 생식소 후단부 한쪽 끝에서 신장된 체세포 돌기가 생식소와 융합되어 작은 난소강이 형성되었다. 부화 후 30일(12.19~13.72 ㎜)에는 생식소 하단부에 정소관 원기가 출현하여 정소의 분화가 시작되었다. 부화 후 50일째 자어(16.28~17.06 ㎜)의 난소내에서는 주변인기 난모세포로 채워지기 시작하였으며, 정소내에서는 정원세포(spermatogonia, SG)의 활발한 증식을 관찰할 수 있었다. 부화 후 250일째 자어(35.49~51.12 ㎜)에서는 난소의 크기가 더욱 커지고 난모세포들로 가득 채워져 있었고, 정원세포의 수가 증가하기 시작하였다. 이상의 결과 눈동자개의 성분화 양상은 자웅이체형 중 분화형이었다.