COLD PRESERVED TESTIS IN NILE TILAPIA LARVAE
KESIMPULAN UMUM
Testis ikan gurami mengandung sel yang kompeten sebagai donor dalam xenotransplantasi menggunakan larva ikan nila sebagai resipien.
SARAN
Untuk mencapai target akhir yaitu memproduksi sel gamet ikan gurami dari ikan nila melalui aplikasi teknologi xenotransplantasi sel testikular maka dibutuhkan uji lanjut terhadap kemampuan sel spermatogonia ikan gurami terdiferensiasi menjadi sel gamet yang fungsional dan mengkaji faktor-faktor yang mempengaruhi perkembangan sel donor pada resipien termasuk mengkaji peluang penggunaan resipien ikan nila triploid sebagai upaya memberikan lingkungan mikro yang maksimum kepada sel donor ikan gurami untuk berkembang dan menghasilkan sel gamet yang fungsional.
Achmad M. 2009. Pengembangan marka molekuler DNA dalam identifikasi sel gonad ikan gurame Osphronemus gouramy dan ikan nila Oreochromis niloticus menggunakan PCR. [Master Tesis]. Bogor : Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor.
Affandi R, Tang M. 2002. Fisiologi Hewan Air. Unri Press.
Aitken RJ, Bajker MA. 2006. Oxidative stress, sperm survival and fertility control. Mol Cell Endocrinol 25:66–69.
Ali HM. 1987. The Histogenesis of lymphoid organs in Tilapia mossambica and antibody responses in adults and fry. [Doctoral Thesis]. Aston University. Almeida FF, Kristoffersen C, Taranger GL, Schulz RW . 2008. Spermatogenesis
in Atlantic cod (Gadus morhua): a novel model of cystic germ cell development. Biol Reprod 78:27–34.
Bellve AR, Milletfe CF, Bhatnagar YM, Obrien DO. 1977. Dissociation of the mouse testis and characterization of isolated spermatogenic cells. J Histochem Cytochem 25:480–494.
Bendsen E, Laursen SB, Olesen C, Westergaard LG, Andersen CY, Byskov AG. 2001. Effect of 4-octylphenol on germ cell number in cultured human fetal gonads. Hum Reprod 16:236–243.
Brinster RL, Zimmermann JW. 1994. Spermatogenesis following male germ cell transplantation. Proc Nat Acad Sci 91:11298–11302.
Browne RK, Clulow J, Mahony M. 2001. Short-term storage of cane toad (Bufo marinus) gametes. Reproduction 121:167–173.
Bryant PJ. 1999. Filopodia: Fickle fingers of cell fate? Curr Biol 9:655–657 Carrasco LAP, Doroshov S, Penman DJ, Bromage N. 1998. Long-term,
quantitative analysis of gametogenesis in autotriploid rainbow trout, Oncorhynchus mykiss. J Reprod Fertil 113:197–210.
Chang IK, Yoshiki A, Kusakabe M, Tajima A, Chikamune T, Naito M, Ohno T. 1995. Germ line chimera produced by transfer of cultured chick primordial germ cells. Cell Biol Int 19:569–576.
Chantanachookhin C, Seikai T, Tanaka M, 1991. Comparative study of the ontogeny of the lymphoid organs in three species of marine fish. Aquaculture 99:143–155.
Choi YJ, Park JK, Lee MS, Ahn JD, Hwang KC, Song H, Kim JH. 2007. Long- term follow-up of porcine male germ cells transplanted into mouse testes. Zygote 15:325–335.
Chong SW, Nguyet LM, Jiang YJ, Korzh V. 2007. The chemokine Sdf-1 and its receptor Cxcr4 are required for formation of muscle in zebrafish. BMC Dev Biol 7:1–14.
Costa DC. 2010. Embryological and micromanipulation techniques in zebrafish (Danio rerio) and Pacific oyster (Crassostrea gigas). [Doctoral Thesis]. Valencia:Universidad Politechnica De Valencia.
Crabbe E, Verheyen G, Tournaye H, Van Steirteghem A. 1997. The use of enzymatic procedures to recover testicular germ cells. Hum Reprod 12:1682–1687.
Daniel JCJr. 1971. Methods in Mammalian Embriology. San Fransisco : W.H. Freeman and company.
De Alvarenga ER, Franca LR. 2009. Effects of different temperatures on testis structure and function, with emphasis on somatic cells, in sexually mature Nile tilapia (Oreochromis niloticus). Biol Reprod 80:537–544.
De Rooij DG, van Diessel-Emiliani FMF. 1997. Regulation of proliferation and differentiation of stem cells in the male germ line. Di dalam Potten CS, editor. Stem Cells. London:Academic. hlm 283–313.
De Rooij DG, Russell LD. 2000. All you wanted to know about spermatogonia but were afraid to ask. J Androl 21:776–798.
Dobrinski I, Avarbock MR, Brinster RL. 1999. Transplantation of germ cells from rabbits and dogs into mouse testes. Biol Reprod 61: 1131–1139.
Dobrinski I. 2005. Germ cell transplantation. Semin Reprod Med 23: 257–265. Doitsidou M, Reichman-Fried M, Stebler J, Koprunner M, Dorries J, Meyer D,
Esguerra CV, Leung T, Raz E. 2002. Guidance of primordial germ cell migration by the chemokine SDF-1. Cell 111:647–659.
Du M, Han H, Jiang B, Zhao L, Qian C, Shen H, Xu Y, Li Z. 2006. An efficient isolation method for domestic hen (Gallus domesticus) ovarian primary follicles. J Reprod Dev 52:570–576.
Dumstrei K, Mennecke R, Raz E. 2004. Signaling pathways controlling primordial germ cell migration in zebrafish. J Cell Sci 117:4787–4795. Dwyer MA, Huang AH, Pan CQ, Lazarus RA. 1999. Expression and
characterization of a DNase I-Fc fusion enzyme. J Biol Chem 274:9738– 9743.
Ehmcke J, Schlatt S. 2008. Focus on fertility preservation : animal models for fertility preservation in the male. Reproduction 136:717–723.
Eriani K, Boediono A, Djuwita I, Sumarsono SH, Al-Azhar. 2008. Development of domestic cat embryo produced by preserved sperms. Hayati J Bio 15:155–160.
Figueiredo JR, Hulshof SC, Van den Hurk R, Ectors FJ, Fontes RS, Nusgens B, Bevers MM, Beckers JF. 1993. Development of a combined new mechanical and enzymatic method for the isolation of intact preantral follicles from fetal, calf and adult bovine ovaries. Theriogenology 40:789– 799.
Fishelson L, Delarea Y, Gon O. 2006. Testis structure, spermatogenesis, spermatocytogenesis, and sperm structure in cardinal fish (Apogonidae, Perciformes). Anat Embryol 211:31–46.
Freshney RI. 2005. Culture of Animal Cells : A Manual of Basic Technique. John Wiley and Sons Inc.
Fujimura K, Okada N. 2007. Development of the embryo, larva and early juvenile of Nile tilapia Oreochromis niloticus (Pisces: Cichlidae). Developmental staging system. Develop Growth Differ 49:301–324. Garcia-Castro MI, Anderson R, Heasman J, Wylie C. 1997. Interactions between
germ cells and extracellular matrix glycoproteins during migration and gonad assembly in the mouse embryo. J Cell Biol 138:471–480.
Grier HJ. 1993. Comparative organization of sertoli cells including the sertoli cell barrier. Di dalam: Russell LD and Griswold MD, editor. The Sertoli Cell. Cache River Press. hlm 704–730.
Griswold MD, McLean D, Russell L. 2001. Promise and limitations of germ cell transplantation in the testis. J Androl 22:713–717.
Heasman J, Hynes RO, Swan AP, Thomas V, Wylie C. 1981. Primordial germ cells of Xenopus embryos: the role of fibronectin in their adhesion during migration. Cell 27:437–447.
Herrid M, Vignarajan S, Davey R, Dobrinski I, Hill JR. 2006. Successful transplantation of bovine testicular cells to heterologous recipients. Reproduction 132:617–624.
Hess RA, Franca LR. 2007. Spermatogenesis and cycle of the seminiferous epithelium. Di dalam: Cheng CY, editor. Molecular Mechanisms in Spermatogenesis. Landes Bioscience. hlm 1–15.
Higashino T, Miura T, Miura C, Higashino KY. 2002. Histological studies on early oogenesis in barfin flounder (Verasper moseri). Zoo Sci 19:557–563. Hill JR, Dobrinski I. 2006. Male germ cell transplantation in livestock. Reprod
Fertil Dev 18:13–18.
Honaramooz A, Yang Y. 2011. Recent advances in application of male germ cell transplantation in farm animals. Vet Med Int 2011:1–9.
Hong Y, Liu T, Zhao H. 2004. Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro. Proc Nat Acad Sci 101:8011–8016.
Horan PK, Melnicoff MJ, Jensen BD, Slezak SE. 1999. Fluorescent cell labeling for in vivo and in vitro cell tracking. Methods Cell Biol 33:469–490.
Jaglarz MK, Howard KR. 1995. The active migration of Drosophila primordial germ cells. Development 121:3495–3503.
Ijiri S, Kaneko H, Kobayashi T, Wang DS, Sakai F, Paul-Prasanth B, Nakamura M, Nagahama Y. 2008. Sexual dimorphic expression of genes in gonads during early sexual differentiation of a teleost fish, the Nile tilapia Oreochromis niloticus. Biol Reprod 78:333–341.
Jahnukainen K, Ehmcke J, Hergenrother SD, Schlatt S. 2007. Effect of cold storage and cryopreservation of immature non-human primate testicular tissue on spermatogonial stem cell potential in xenografts. Hum Reprod 22:1060–1067.
Jiang FX , Short RV. 1998. Male germ cell transplantation: present achievements and future prospects. Int J Dev Biol 42:1067–1073.
Johnston DS, Russel LD, Griswold MD. 2000. Advances in spermatogonial stem cell transplantation. Rev Reprod 5:183–188.
Jones PA, Werb Z. 1980. Degradation of connective tissue matrices by macrophages : influence of matrix composition on proteolysis of glycoproteins, elastin, and collagen by macrophages in culture. J Exp Med 152:1527–1536.
Kiernan JA. 1990. Histological and Histochemical Methods : Theory and Practice. Toronto: Pergamon Press.
Kim Y, Selvaraj V, Dobrinski I, Lee H, Mcentee MC,Travis AJ. 2006. Recipient preparation and mixed germ cell isolation for spermatogonial stem cell transplantation in domestic cats. J Androl 27:248–256.
Kobayashi T, Kajiura-Kobayashi H, Nagahama Y. 2000. Differential expression of vasa homologue gene in the germ cells during oogenesis and spermatogenesis in a teleost fish, tilapia, Oreochromis niloticus. Mech Dev 99:139–142.
Kobayashi T, Takeuchi Y, Takeuchi T, Yoshizaki G. 2007. Generation of viable fish from cryopreserved primordial germ cells. Mol Reprod Dev 74:207– 213.
Kobayashi T. 2010. In vitro germ cell differentiation during sex differentiation in a teleost fish. Int J Dev Biol 54:105–111.
Kuske CR, Banton KL, Adorada DL, Stark PC, Hill KK, Jackson PJ. 1998. Small-scale DNA sample preparation method for field PCR detection of microbial cells and spores in soil. Appl Environ Microbiol 64:2463–2472. Lacerda SMSN, Batlouni SR, Assis LH, Resende FM, Campos-Silva SM,
Campos-Silva R, Segatelli TM, França LR. 2008. Germ cell transplantation in tilapia (Oreochromis niloticus). Cybium 32:115–118.
Lahnsteiner F, Weismann T, Patzner RA. 1997. Aging processes of rainbow trout semen during storage . The Progress Fish-Cult 5:272–279.
Leal MC, Cardoso ER, Nobrega RH, Batlouni SR, Bogerd J, França LR, Schulz RW. 2009. Histological and stereological evaluation of zebrafish (Danio rerio) spermatogenesis with an emphasis on spermatogonial generations. Biol Reprod 81:177–187.
Loir M, Sourdaine P, Mendis-Handagama SMLC, Jegou B. 1995. Cell-cell interactions in the testis of teleost and elasmobranchs. Micr Res Tech 32:533–552.
Majhi SK, Hattori RS, Yokota M, Watanabe S, Strussmann CA. 2009. Germ cell transplantation using sexually competent fish: an approach for rapid propagation of endangered and valuable germline. PLoS ONE 4:e6132. doi:10.1371/journal.pone.0006132.
Manning MJ, Nakanishi T. The specific immune : celluler defences. Di dalam: Iwama G, Nakanishi T, editor. The Fish Immune System. New York: Academic Press. hlm 159–205.
Marret C, Durand P. 2000. Culture of porcine spermatogonia: effects of purification of the germ cells, extracellular matrix, and fetal calf serum on their survival and multiplication. Reprod Nut Dev 40:305–319.
Mather JP, Roberts PE. 1998. Introduction to Cell and Tissue Culture: Theory and Technique. New York: Plenum Press.
Meachem S, Schonfeldt V, Schlatt S. 2001. Spermatogonia: stem cells with a great perspective. Reproduction 121:825–834.
Miura T. 1999. Spermatogenic cycle in fish. Di dalam: Knobil E, Neill JD, editor. Encyclopedia of Reproduction. Vol 4. San Diego: Academic Press. hlm 571–578.
Mulero I, Garcia-Ayala A, Meseguer J, Mulero V. 2007. Maternal transfer of immunity and ontogeny of autologous immunocompetence of fish: a minireview. Aquaculture 268:244-250.
Nagahama Y. 1994. Endocrine regulation of gametogenesis in fish. Int J Dev Biol 38:217–229.
Nagano M, Avarbock MR, Brinster RL. 1999. Pattern and kinetics of mouse donor spermatogonial stem cell colonization in recipient testes. Biol Reprod 60:1429–1436.
Nagano M, McCarre JR, Brinster RL. 2001. Primate spermatogonial stem cell colonize mouse testes. Biol Reprod 64:1409–1416.
Nakamura M, Kobayashi T, Chang XT, Nagahama Y. 1998. Gonadal sex differentiation in teleost fish. J Exp Zool 281:362–372.
Nicosia SV, Evangelista I, Batta SK. 1975. Rabbit ovarian follicles. I. Isolation technique and characterization at different stage of development. Biol Reprod 13:423–447.
Nobrega RH, Batlouni SR, Franca LR. 2009. An overview of functional and stereological evaluation of spermatogenesis and germ cell transplantation in fish. Fish Physiol Biochem 35:197–206.
Oatley JM, Avarbock MR, Telaranta AI, Fearon DT, Brinster RL. 2006. Identifying genes important for spermatogonial stem cell self-renewal and survival . Proc Nat Acad Sci 103:9524–9529.
Oatley JM, Oatley MJ, Avarbock MR, Tobias JW, Brinster RL. 2009. Colony stimulating factor 1 is an extrinsic stimulator of mouse spermatogonial stem cell self-renewal. Development 136:1191–1199.
Ogawa T, Dobrinski I, Avarbock MR, Brinster RL. 1999. Xenogenic spermatogenesis following transplantation of hamster germ cell to mouse testes. Biol Reprod 60:515–521.
Ohta H, Yomogida K, Dohmae K, Nishimune Y. 2000. Regulation of proliferation and differentiation in spermatogonial stem cells: the role of c- kit and its ligand SCF. Development 127:2125–2131.
Okutsu T, Takeuchi Y, Yoshizaki G. 2006a. Manipulation of fish germ cell : visualization, cryopreservation and transplantation. J Reprod Dev 52:685– 693.
Okutsu T, Suzuki K, Takeuchi Y, Takeuchi T, Yoshizaki G. 2006b. Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish. Proc Nat Acad Sci 103:2725–2729.
Okutsu T, Shikina S, Kanno M, Takeuchi Y, Yoshizaki G. 2007. Production of trout offspring from triploid salmon parents. Science 317:1517.
Olive V, Cuzin F. 2005. The spermatogonial stem cell: from basic knowledge to transgenic technology. The Int J Biochem Cell Biol 37:246–250.
Pieterse GM. 2004. Histopathological changes in the testes of Oreochromis mossambicus (Cichlidae) as a biomarker of a heavy metal pollution.[Doctoral Thesis]. Johannesburg : Rand Africaan University. Promega. 2009. MagneSil® Genomic, Large Volume System. Technical Bulletin. Pudney J. 1993. Comparative cytology of the non-mammalian vertebrate sertoli
cell. Di dalam: Russell LD, Griswold MD, editor. The Sertoli Cell. Clearwater, FL: Cache River Press. hlm611–657.
Roche. 2003. Cell proliferation and viability measurement. Biochemica 3:26–29. Saanin H. 1984. Taksonomi dan Kunci Identifikasi Ikan. Jilid II. Binacipta.
Bandung.
Saito T, Goto-Kazeto R, Arai K, Yamaha E. 2008. Xenogenesis in teleost fish through generation of germ-line chimeras by single primordial germ cell transplantation. Biol Reprod 78:159–166.
Santos JE, Bazzoli N, Rizzo E, Santos GB .2001. Morphofunctional organization of the male reproductive system of the catfish Iheringichthys labrosus (Luetken, 1874) (Siluriformes:Pimelodidae). Tisues and Cell 33:533–540. Schatt S. 2002. Spermatogonial stem cell preservation and transplantation. Mol
Schultz RW, Miura T. 2002. Spermatogenesis and its endocrine regulation. Fish Physio Biochem 26:43–56.
Schulz RW, Menting S, Bogerd J, Franca LR, Vilela DAR, Godinho HP. 2005. Sertoli cell proliferation in the adult testis – evidence from two fish species belonging to different orders. Biol Reprod 73:891–898.
Segatelli TM, Franca LR, Pinheiro PFF, Almeida CCD, Martinez M, Martinez FE. 2004. Spermatogenic cycle length and spermatogenic efficiency in the gerbil (Meriones unguiculatus). J Androl 25:872–880.
Sheetz MP. 1979. DNase I-dependet dissociation of erythrocyte cytoskeletons. Cell Bio 81:266–270.
Shinohara T, Brinster LR. 2000. Enrichment and transplantation of spermatogonial stem cells. Int J Androl 23:89–91.
Sikka SC. 1996. Oxidative stress and role of antioxidants in normal and abnormal sperm function. Front in Biosci 1:78–86.
Stickney R. 2000. Tilapia culture. Di dalam Stickney RR, editor. Encyclopedia of Aquaculture. New York: John Wiley & Sons. hlm 83.
Sofikitis N, Mio Y, YamamotoY, Miyagawa I. 1999. Transplantation of human spermatogonia into the seminiferous tubules of animal testicles result in the completion of the human meiosis and the generation of human motile spermatozoa. Fertil Steril 72:83–89.
Suresh R, Aravindan GR, Moudgal VR. 1992. Quantitation of spermatogenesis by DNA flow cytometry: comparative study among six species of mammals. J Biosci 17:413–419.
[SNI] Standardisasi Nasional Indonesia. 2000. Induk ikan gurame (Osphronemus gouramy, Lac) kelas induk pokok (parent stock). SNI-01-6485.1-2000. Tajima A, Naito M, Yasuda Y, Kuwana T. 1993. Production of germ line
chimera by transfer of primordial germ cells in the domestic chicken (Gallus domesticus). Theriogenology 40:509–519.
Takashima F, Hibiya F. 1995. An Atlas of Fish Histology : Normal and Pathological Features. Tokyo: Kodansha Ltd.
Takeuchi Y , Yoshizaki G, Kobayashi T, Takeuchi T. 2002. Mass isolation of primordial germ cells from transgenic rainbow trout carrying the green fluorescent protein gene driven by the vasa gene promoter. Biol Reprod 67:1087–1092.
Takeuchi Y, Yoshizaki G, Takeuchi T. 2003. Generation of live fry from intraperitonially transplantation primordial germ cells in rainbow trout. Biol Reprod 6:1142–1149.
Takeuchi Y, Takeuchi T , Yoshizaki G. 2004. Surrogate broodstock produces salmonids. Nature 430 : 630–639.
Takeuchi Y, Higuchi K, Yatabe T, Miwa M, Yoshizaki G. 2009. Development of spermatogonial cell transplantation in nibe croaker, Nibea mitsukurii (Perciformes, Sciaenidae). Biol Reprod 81:1055–1063.
Vilela DAR, Silva SGB, Peixoto MTD, Godinho HP, Franca LR. 2003. Spermatogenesis in teleost: insights from the Nile tilapia (Oreochromis niloticus) model. Fish Phy Biochem 28:187–190.
Wallace PK, Tario JD Jr, Fisher JL, Wallace SS, Ernstoff MS, Muirhead KAPK. 2008. Tracking antigen-driven responses by flow cytometry: monitoring proliferation by dye dilution. Cytometry 73A:1019–1034.
Weidinger G, Wolke U, Koprunner M, Thisse C, Thisse B, Raz E. 2002. Regulation of zebrafish primordial germ cell migration by attraction towards an intermediate target. Development 129:25–36.
Wong T, Saito T, Crodian J, Collodi P. 2011. Zebrafish germline chimeras produced by transplantation of ovarian germ cells into sterile host larvae. Biol Reprod 84:1190–1197.
Worthington. 2003. Tissue Dissociation Guide. Worthington Biochemical Corporation.
Yamamoto, T. 1983. Sex differentiation. Di dalam: Hoar WS, Randall DJ, editor. Fish Physiology. Vol. 3. New York: Academic Press. hlm 117−175.
Yano A, Suzuki K, Yoshizaki G. 2008. Flow-cytometric isolation of testicular germ cells from rainbow trout (Oncorhynchus mykiss) carrying the green fluorescent protein gene driven by trout vasa regulatory regions. Biol Reprod 78:151–158.
Yazawa M, Ferrante C, Feng J, Mio K, Ogura T, Zhang M, Lin P, Pan Z, Komazaki S, Kato K, Nishi M, Zhao X, Weisleder N, Sato C, Ma J, Takeshima H. 2007. TRIC channels are essential for Ca2+ handling in intracellular stores. Nature 448:78–83.
Yazawa R, Takeuchi Y, Higuchi K, Yatabe T, Kabeya N, Yoshizaki G. 2010. Chub mackerel gonads support colonization, survival, and proliferation of intraperitoneally transplanted xenogenic germ cells.Biol Reprod 82:896– 904.
Yoshizaki G, Okutsu T, Ichikawa M, Hayashi M, Takeuchi Y. 2010. Sexual plasticity of rainbow trout germ cells. Anim Reprod 7:187–196.
Young G, Kusakabe M, Nakamura I. 2005. Gonadal steroidogenesis in teleost fish. Di dalam: Melamed P, Sherwood N, editor. Hormones and Their Receptors in Fish Reproduction. Singapore: World Scientific. hlm 155-223. Zapata RF. 2009. Expression of gfp in transgenic tilapia under the control of the
medaka β – actin promoter : establishment of a model system for germ cell transplantation. [Master Thesis]. Tokyo : Master’s Course of Marine Life Sciences, Tokyo University of Marine Science and Technology.
Zhou Q, Griswold MD. 2008. Regulation of spermatogonia. The Stem Cell Research Community. StemBook. doi/10.3824/ stembook.1.7.1. http://www.stembook.org/index.html [14 Juli 2008].
Lampiran 1 Analisis karakteristik morfologi tipe spermatogonia ikan gurami
ANOVA (lanjutan)
F Sig,
Diameter sel Between Groups 712,245 ,000 Diameter inti Between Groups 112,315 ,000 Volume Sel Between Groups 394,925 ,000 Volume inti Between Groups 74,601 ,000 Volume sitoplasma Between Groups 427,841 ,000
Post Hoc Tests
Diameter inti
Tipe sel
Subset for alpha = 0,05
N 1 2 3 Duncana SpB 90 5,9222 SpT 90 8,3250 SpA 90 8,3833 SSC 90 8,7917 Sig, 1,000 ,738 1,000 ANOVA
Sum of Squares Mean Square
Diameter sel Between Groups 4883,242 3 1627,747
Within Groups 813,594 356 2,285
Total 5696,836 359
Diameter inti Between Groups 460,171 3 153,390
Within Groups 486,193 356 1,366
Total 946,364 359
Volume Sel Between Groups 5,043E8 3 1,681E8
Within Groups 1,515E8 356 425617,000
Total 6,558E8 359
Volume inti Between Groups 3404584,990 3 1134861,663
Within Groups 5415640,978 356 15212,475
Total 8820225,968 359
Volume sitoplasma Between Groups 4,406E8 3 1,469E8
Within Groups 1,222E8 356 343270,224
Volume Sel
Tipe sel
Subset for alpha = 0,05
N 1 2 3 4 Duncana SpB 90 394,1871 SpT 90 1008,1969 SpA 90 2185,6430 SSC 90 3489,5740 Sig, 1,000 1,000 1,000 1,000 Volume inti Tipe sel
Subset for alpha = 0,05
N 1 2 3 Duncana SpB 90 121,2708 SpT 90 316,9541 SpA 90 331,2899 SSC 90 373,6576 Sig, 1,000 ,436 1,000 Volume sitoplasma Tipe sel
Subset for alpha = 0,05
N 1 2 3 4 Duncana SpB 90 272,9163 SpT 90 691,2428 SpA 90 1854,3531 SSC 90 3115,9164 Sig, 1,000 1,000 1,000 1,000
Lampiran 2 Analisis data kelimpahan sel testikular (frekuensi relatif) ikan gurami pada bobot tubuh ikan <500 g (1), 500–1000 g (2), >1000 g (3)
Bobot tubuh (g) SSC SpA SpT SpB SL
1. <500 1 4,17 16,67 14,58 13,54 41,67 1 6,00 18,00 15,00 6,00 54,00 1 4,00 20,00 14,00 16,00 47,00 1 4,30 13,98 9,68 0,00 41,94 1 2,06 15,46 9,28 4,12 55,67 1 3,09 4,12 8,25 2,06 55,67 1 5,21 12,50 9,38 3,13 47,92 1 3,09 17,53 15,46 14,43 43,30 1 3,53 14,12 10,59 3,53 54,12 2. 500-1000 2 3,03 19,19 19,19 15,15 46,46 2 2,00 18,00 14,00 8,00 43,00 2 2,00 22,00 29,00 31,00 41,00 2 4,12 22,68 22,68 16,49 45,36 2 2,06 26,80 18,56 14,43 41,24 2 5,10 30,61 15,31 22,45 43,88 2 2,06 23,71 16,49 19,59 24,74 2 4,17 21,88 20,83 15,63 45,83 2 2,11 24,21 14,74 11,58 40,00 3. >1000 3 0,00 6,82 19,32 15,91 51,14 3 1,02 4,08 10,20 12,24 17,35 3 0,00 8,00 18,00 10,00 52,00 3 0,00 5,00 18,00 16,00 70,00 3 1,00 8,00 17,00 18,00 50,00 3 0,00 10,00 15,00 18,00 40,00 3 0,00 5,10 14,29 16,33 52,04 3 0,00 11,00 9,00 14,00 40,00 3 1,03 6,19 16,49 16,49 43,30 Keterangan : SSC : sel punca spermatogonia, SpA : spermatogonia A,
SpT : spermatogonia transisi, SpB : spermatogonia B, Sel L: sel derivat spermatogonia dan sel somatik
ANOVA
Sum of Squares df Mean Square F Sig,
SSC Between Groups 62,376 2 31,188 30,136 0,000
Within Groups 24,838 24 1,035
Total 87,214 26
SpA Between Groups 1167,625 2 583,813 42,658 0,000
Within Groups 328,458 24 13,686 Total 1496,083 26 SpT Between Groups 231,807 2 115,904 7,971 0,002 Within Groups 348,991 24 14,541 Total 580,798 26 SpB Between Groups 524,993 2 262,496 8,994 0,001 Within Groups 700,450 24 29,185 Total 1225,442 26
Sel lain Between Groups 277,101 2 138,550 1,497 0,244
Within Groups 2221,243 24 92,552
Total 2498,344 26