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1 14 1 Vol14 No Life Science Research Feb , 2 2*, : (Hynobius guabangshanensis) : 1) : ;, ; 2),,,, ; 3) (1941 ± 715 μm), : (2298%), (6579 %) : ; ; ; : Q954 : A : (10) 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 Hunan China 2 College of Life Sciences Hunan Normal University Changsha 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 : : : 05JJ30068 Life Science Research ~043 Hynobius guabangshanensis Amphibian Urodela Hynobiidae Hynobius 700 m : * Tel dxj @163com

2 1 : 39 2 % ph = % [1,2] S540 2 % [3,4] r / min 1 % Epon812 H SPSS % 5 % % 1 a b 10~24 h - H-E ± 715 μm Olympus ± 697 μm MapInfo70 Kuramoto ± 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 ± ± ± ± ± 28 9 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0 37 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± * [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

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4 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 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 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 neck piece h i nuclear fossa proximal centriole acosome distal centriole perforatorium endonuclear canal nucleus 1 r h i

5 42 10 marginal fibre 32 Picheral 1979 Kuramoto Kim et al [3,4,7,10,11] 1 l q 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 ± 0935 μm Hynobius nebulosus H kimurae 1941 ± 715 μm H takedai 1810 % % [5] 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 ± 550 μm ± 390 takedai H lichenatus H μm 2727 ± 550 retardatus H stejneger [7] [7] ± 870 μm Mori H leechii [4] Batrachuperus tibe tanus [ 8] Batrachuperus longdon- gensis [9,10] : Mistsuru Kuramoto (Fukuoka University of Education) Daesik Park(Kangwon National University)

6 1 : 43,! (References): [2] [J] DENG Xue-jian SHEN You-hui WANG Bin Hynobius guabangshanensis[j] China Nature KURAMOTO M Scanning electron microscopic studies on the spermatozoa of some Japanese salamanders Hynobiidae Cryptobranchidae Salamandridae [J] Japanese Journal of Herpetology [4] KIM K H PARK W H LEE Y H Ultrastructure of spermatozoa in urodeles Hynobius leechii Amphibia Urodela [J] Korean J Electron Microscopy [5] MORI K Ueber die Struktur der Samenfaden bei einigen Urodelen[J] J Med Soc Nagasaki [6] KERA Y Spermatozoa of Onychodactylus japonicus [J] Nippon Herpetol Journal [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 [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 [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 [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 [11] PICHERAL B Structural comparative and functional aspects of spermatozoa in urodeles [C]//FAWCETT D W BEDFORD J M The spermatozoon

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