14 1 Vol14 No1 10 2 Life Science Research Feb 10 10 1, 2 2*, 1 410014 2 410081 : (Hynobius guabangshanensis) : 1) : ;, ; 2),,,, ; 3) (1941 ± 715 μm), : (2298%), (6579 %) : ; ; ; : Q954 : A : 1007-7847(10)01-0038-06 Morphological Structure of the Spermatozoa in Hynobius guabangshanensis Urodela: Hynobiidae GUO Ke-ji 1 MI Xiao-qi 2 DENG Xue-jian 2* 1 South-Central Forestry Inventory Planning & Designing Institute of State Forestry Administration Changsha 410014 Hunan China 2 College of Life Sciences Hunan Normal University Changsha 410081 Hunan China Abstract The ultrastructure of the spermatozoa of Hynobius guabangshanensis was studied by both of the scanning and transmission electron microscopy methods The results showed that 1 the spermatozoa of Hynobius guabangshanensis share some characteristic morphological features with that of Hynobiidae eg thin and straight head taper acrosome very short and unconspicuous neck relatively long tail with undulating membrane 2 their spermatozoa s ultrastructure also share some synapomorphies with that of Hynobiidae eg the clover-shaped acrosome without acrosomal barb absence of mitochondrion in the tail and cylindric axial fibre 3 in additionally except for the medium size total length = 1941 ± 715 μm in Hynobiidae the morphological statistics of Hynoius guabangshanensis s sperm differ significantly from other species of Hynobiidae the proportion of head length to total length 2298 % is relatively higher and the proportion of tail length 6579 % is much lower than that of others Key words Hynobius guabangshanensis spermatozoa morphology ultrastructure : 09-03-18 : 09-06-28 : 05JJ30068 Life Science Research 10 14 1 038~043 Hynobius guabangshanensis Amphibian Urodela Hynobiidae Hynobius 700 m 04 04 : 1981- * 1953- Tel 0731-88871922 E-mail dxj8871922@163com
1 : 39 2 % ph = 72 3 1% [1,2] S540 2 % [3,4] 1 000 r / min 1 % Epon812 H-600 13 1 11 SPSS 05 12 4 2 21 2 % 5 % 12 121 5 % 1 a b 10~24 h - H-E 1 1941 ± 715 μm Olympus 122 1277 ± 697 μm MapInfo70 Kuramoto 1995 218 ± 0997 μm 446 ± 0935 μm 1 Table 1 The sperm size of spermatozoa of 16 Hynobiidae species (Mean±SD in μm) Species Length Acrosome / μm % Head / μm % Tail / μm % Total / μm n References H guabangshanensis H nebulosus H dunni H tsuensis H nigrescens H naevius H takedai H lichenatus H retardatus H stejnegeri H leechii P boulengeri S keyserlingii O japonicus B tibetanus B Longdongensis 218 ± 099 181 ± 210 217 ± 150 8 ± 060 2 ± 28 9 ± 1 181 ± ± 14 176 ± 1 224 ± 140 178 ± 130 50 166 ± 140 171 ± 080 144 ± 080 180 ± 369 136 ± 152 1707 1584 1455 1440 1237 1288 1801 1643 07 1451 1369 1415 1328 427 992 941 446 ± 094 456 ± 230 790 ± 290 745 ± 190 465 ± 23 895 ± 240 504 ± 1 546 ± 2 603 ± 240 671 ± 310 12210 627 ± 0 665 ± 0 37 ± 490 908 ± 833 1017 ± 510 2298 2563 3163 3108 2848 3282 2982 3045 3103 3232 4183 3189 3131 3705 3134 3921 1277 ± 697 1143 ± 470 1491 ± 1030 1444 ± 390 966 ± 37 1623 ± 460 1005 ± 370 1071 ± 330 1116 ± 310 1227 ± 5 1497 1173 ± 460 1288 ± 440 3372 ± 1660 1811 ± 1153 1444 ± 353 6579 6425 5969 6024 5915 5952 5947 5973 5744 5910 5130 5966 6064 6133 6247 5567 1941 ± 715 1779 ± 550 2498 ± 870 2397 ± 410 1634 ± 39 2727 ± 550 1690 ± 390 1793 ± 400 1943 ± 390 76 ± 630 29180 1966 ± 540 2124 ± 5 5498 ± 1970 2899 ± 1302 2594 ± 482 30 32 1 1 1 1-15 15 * [4] [7] [7] [7] [9] [8] H leechii n: ; *: Notes Percentage of the Table 1 is the parts of sperm length and the ratio of full-length the number of measured samples of H leechii is unknown n Number of samples *: This experiment
40 10
1 : 41 1 a), ( 400); (b), ( 1 000); (c), =10 μm; (d), = 1 μm; (e), =2 μm; (f), = 1 μm; (g),, =1 μm; (h),, =02 μm; (i),, = 02 μm; (j),, = 02 μm; (k), = 01 μm; (l),, = 02 μm; (m) (a) (b),, = 02 μm; (n),, = 05 μm; (o),, = 02 μm; (p),, = 02 μm; (q), = 02 μm; (r),, = 02 μm A: ;AF: ;AX: ;C: ;DC: ;EC: ;H: ;M: ;Mi: ;MF: ; N: ;NE: ;NF: ;NP: ;NR: ;PC: ;Pe: ;T: ;UM: Fig1 Microstructure and ultrastructure of the spermatozoa of Hynobius guabangshanensis a LM photograph of spermatozoa showing head tail and undulating membrane 400 b LM photograph of spermatozoa showing head tail and undulating membrane 1 000 c SEM photograph of spermatozoa scale bar = 10 μm d SEM photograph of the anterior end of the head showing no acrosomal barb scale bar = 1 μm e SEM photograph of the posterior end of the head scale bar=2 μm f SEM photograph of the neck piece scale bar=1 μm g SEM photograph of the tail showing axial fibre axoneme undulating membrane scale bar = 1 μm h Cross section of the anterior end of the head showing acrosome perforatorium nucleus scale bar=02 μm i Cross section of the anterior end of the head showing acrosome perforatorium nucleus scale bar = 02 μm j Cross section of the anterior end of the head showing acrosome perforatorium nucleus scale bar = 02 μm k Cross section of the head showing nucleus nuclear ridge scale bar = 02 μm l Cross section of the principal piece a and the end piece b of the tail showing axial fibre undulating membrane axoneme marginal fibre scale bar = 02 μm m Cross section of the middle piece of the tail showing axial fibre undulating membrane cortex medulla axoneme marginal fibre scale bar = 02 μm n Cross section of the principal piece of the tail showing axial fibre undulating membrane axoneme and marginal fibre scale bar = 05 μm o Cross section of the principal piece of the tail showing undulating membrane axial fibre marginal fibre scale bar = 02 μm p Longitudinal section of the head showing nucleus endonuclear canal scale bar = 02 μm q Longitudinal section of the middle piece of the tail showing cortex and medulla scale bar = 02 μm r Longitudinal section of the neck piece showing nucleus nuclear fossa distal centriole and proximal centriole scale ba=02 μm A acrosome AF axial fibre AX axoneme C cortex DC distal centriole EC endonuclear canal H head M medulla Mi mitochondrion MF marginal fibre N nucleus NE nuclear membrane NF nuclear fossa NP neck piece NR nuclear ridge PC proximal centriole Pe perforatorium T tail UM undulating membrane 22 221 1 p 1 p acrosomal barb 1 c d e f axial 1 r fibre undulating membrane nuclear ridge 1 j k axoneme 1 c g p 1 k 222 2222 neck piece 2221 1 h i nuclear fossa proximal centriole acosome distal centriole perforatorium endonuclear canal nucleus 1 r 2233 1 h i
42 10 marginal fibre 32 Picheral 1979 Kuramoto 1995 1997 Kim et al 1995 05 [3,4,7,10,11] 1 l q 9+2 9 1 l m n o 1 l 33 1 m -a n 1 1 m -a Pachypalaminus boulengeri 1966 ± 540 μm [3,7] 1 m -b 1634 ±39 μm 2240 ± 140 μm 3 [4] 1707 % 31 Mori 1936 446 ± 0935 μm Hynobius nebulosus H kimurae 1941 ± 715 μm H takedai 1810 % 07 2298 % [5] 37380 μm [5] Kera 1977 Onychodactylus japonicus 1277 ± 697 μm [6] H dunni 6579 % H tsuensis H nigrescens Kuramoto H naevius O japonicus Pachypalaminus boulengeri Salamandrella keyserlingii [ 3 ] H 17790 ± 550 μm 16340 ± 390 takedai H lichenatus H μm 2727 ± 550 retardatus H stejneger [7] [7] 24980 ± 870 μm Mori H leechii [4] Batrachuperus tibe- 1936 tanus [ 8] Batrachuperus longdon- gensis [9,10] : Mistsuru Kuramoto (Fukuoka University of Education) Daesik Park(Kangwon National University)
1 : 43,! (References): 50 2 9-215 [2] [J] DENG Xue-jian SHEN You-hui WANG Bin Hynobius guabangshanensis[j] China Nature 04 5 18- KURAMOTO M Scanning electron microscopic studies on the spermatozoa of some Japanese salamanders Hynobiidae Cryptobranchidae Salamandridae [J] Japanese Journal of Herpetology 1995 16 2 49-58 [4] KIM K H PARK W H LEE Y H Ultrastructure of spermatozoa in urodeles Hynobius leechii Amphibia Urodela [J] Korean J Electron Microscopy 1995 25 111-121 [5] MORI K Ueber die Struktur der Samenfaden bei einigen Urodelen[J] J Med Soc Nagasaki 1936 14 2 125-132 [6] KERA Y Spermatozoa of Onychodactylus japonicus [J] Nippon Herpetol Journal 1977 9 5-6 [7] KURAMOTO M Further Studies on sperm morphology of Japanese Salamders with special reference to geographic and individual variation in sperm size[j] Japanese Journal of [1] Herpetology 1997 17 1 1-10 [J] SHEN You-hui [8] [J] DENG Xue-jian WANG Bin A new hynobiid species QIN Li-mei XIE Feng ZHENG Zhong-hua et al Hynobius guabangshanensis from Hunan Province China Sperm morphology of alpine stream salamander Batrachuperus Amphibia:Hynobiidae [J] Acta Zoologica Sinica 04 tibetanus[j] Chinese Journal of Zoology 05 40 2 71-74 [9] [J] ZHENG Zhong-hua JIANG Jian-ping XIE Feng et al Sperm morphology of Longdong stream salamander Batrachuperus longdongensis Urodela Hynobiidae [J] Herpetological Research 02 9 4-9 [10] [J] ZHENG Zhong-hua JIANG Jian-ping XIE Feng Ultrastructure of the spermatozoa of Longdong stream salamander Batrachuperus longdongensis Urodela Hynobiidae [J] Acta Zoologica Sinica 05 51 4 703-709 [11] PICHERAL B Structural comparative and functional aspects of spermatozoa in urodeles [C]//FAWCETT D W BEDFORD J M The spermatozoon 1979 267-287