Mahalingam ANBARASU, Ramangalingam KIRUBAGARAN, Gopal DHARANI, Thanumalaya SUBRAMONIAM

Probing the architecture of testis and morphology of male germinal cells in the mud crab with the atomic force microscopy

The architecture of testicular lobules, morphology of spermatozoa, and spermatophore in the mud crab, Scylla serrata, are described for the first time employing atomic force microscopy (AFM). A fast and simple squash preparation protocol was followed using crustacean physiological saline without distorting the natural structure. Observations provided new insight into the arrangement of lobules and morphology of testicular assembly. It consists of a close network of lobules of different sizes in a radial manner on radially branching basal tubules. Budding of new lobules is another distinguishable feature noted in the network of testicular organization. A thin membranous envelope of the lobule with undulating processes offered indirect evidence on the possible storage of sperm cells within the lumen of the lobule. Spermatozoa of S. serrata were of different shapes and sizes, and, depending on the maturational status, they displayed changes in the acrosome vesicle with a significant amount of cytoplasm. Vertical constrictions found in the testicular lobules suggest propulsive extrusion of spermatophoric masses into the central lumen. These observations necessitate further study using AFM and involving molecular probes to elucidate the polymorphic nature of male germinal cells, mechanisms of spermatophore formation, and the phylogenetic relationship among crustaceans in general.

Anahtar Kelimeler:

Key words: Testis, spermatozoa, spermatophore, Scylla serrata, AFM

Probing the architecture of testis and morphology of male germinal cells in the mud crab with the atomic force microscopy

Keywords:

Key words: Testis, spermatozoa, spermatophore, Scylla serrata, AFM,

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Subramoniam T. Spermatophore and sperm transfer in marine crustaceans. Adv Mar Biol 29: 129–214, 1993.
Retzius G. Die Spermien der Crustaceen. Biol Untersuch 14: 1–54, 1909.
Fasten N. Male reproductive organs of decapoda, with special reference to Puget Sound forms. Puget Sound Mar Stat Publ 1: 285–307, 1917.
Dhillon B. Sperm nucleus of Clibanarius longitarsus. Experientia 20: 505–506, 1964.
Pochon-Masson J. L’ultrastrucure des épines du spermatozoïde chez les Décapodes (Macroures, Anomoures, Brachyoures). Comptes Rendus Hebdomadaire des Séances de l’Academie des Sciences, Paris 260: 3762–3764, 1965.
Chevaillier P. Structure et constitution cytochimique de la capsule du spermatozoïde des Crustacés Décapodes. Comptes Rendus Hebdomadaire des Séances de l’Academie des Sciences, Paris 262: 1546–1549, 1966.
Chevaillier P. Nouvelles observations sur la structure des fibres intra-nucléaires du spermatozoïde du Pagure Eupagurus bernhardus L. (Crustacé, Décapode). J de Microscopie 6: 853– 856, 1967.
Tudge CC. Spermatophore diversity within and among the hermit crab families, Coenobitidae, Diogenidae, and Paguridae (Paguroidea, Anomura, Decapoda). Biol Bull 181: 238–247, 19 Tudge CC. Ultrastructure and phylogeny of anomuran crab spermatozoa, PhD, Department of Zoology, The University of Queensland, Australia; 1995.
Tudge CC. Spermatophore morphology and spermatozoal ultrastructure of the recently described hermit crab Strigopagurus boreonotus Forest, 1995 (Decapoda, Anomura, Diogenidae). Bull Mus Nation His Nat Paris 18: 547–555; 1996.
Tudge CC. Phylogeny of Anomura (Decapoda, Crustacea): Spermatozoa and spermatophore morphological evidence. Contrib Zool 67: 125–141; 1997.
Tudge CC. Spermatophore morphology in the hermit crab families Paguridae and Parapaguridae (Paguroidea, Anomura, Decapoda). Invetebr Reprod Dev 35: 203–214; 1999.
Tudge CC. Ultrastructure of the spermatophore lateral ridge in hermit crabs (Decapoda, Anomura, Paguroidea). Crustaceana 72: 77–84; 1999.
Tudge CC. Endemic and enigmatic: the reproductive biology of Aegla (Crustacea: Anomura: Aeglidae) with observations on sperm structure. Memoir Mus Victoria 60: 63–70; 2003.
Allen MJ, Lee JD 4th, Lee C et al. Extent of sperm chromatin hydration determined by atomic force microscopy. Mol Reprod Dev 45: 87–92; 1996.
Lee JD, Allen MJ, Balhorn R. Atomic force microscopy analysis of chromatin volumes in human sperm with head-shape abnormalities. Biol Reprod 56: 42–49; 1997.
Allen MJ, Bradbury EM, Balhorn R. The natural subcellular surface structure of the bovine sperm cell. J Struct Biol 114: 197–208; 1995.
Saeki K, Sumitomo N, Nagata Y et al. Fine surface structure of bovine acrosome: intact and reacted spermatozoa observed by atomic force microscopy. J Reprod Develop 51: 293–298; 2005. Allen MJ, Bradbury EM, Balhorn R. The chromatin structure of well-spread demembranated human sperm nuclei revealed by atomic force microscopy. Scanning Microscopy 10: 989– 996; 1996.
Ellis DJ, Shadan S, James PS et al. Post-testicular development of a novel membrane substructure within the equatorial segment of ram, bull, boar, and goat spermatozoa as viewed by atomic force microscopy. J Struct Biol 138: 187–198; 2002.
Soon LL, Bottema C, Breed WG. Atomic force microscopy and cytochemistry of chromatin from marsupial spermatozoa with special reference to Sminthopis crassicaudata. Mol Reprod Dev 48: 367–374; 1997.
Yamagishi H, Ebara A. Spontaneous activity and pacemaker property of neurones in the cardiac ganglion of an isopod crustacean, Ligia exotica. Comp Biochem Physiol 81A: 55–62; 19 Hinsch GW. Ultrastructure of sperm and spermatophores of the golden crab Geryon fenneri and a closely related species, the red crab G. quinqendens, from the eastern Gulf of Mexico. J Crustacean Biol 8: 340–345; 1988.
Sheeba JR, Sasikala SL, Kirubagaran R et al. Role of serotonin on testicular maturation in the male mud crab, Scylla serrata (Forskal, 1775). Proceedings of XXII Symposium on Reproductive Biology and Comparative Endocrinology held at University of Madras, Chennai, India; 2004.
Helen SB, Jamila PE, Kirubagaran R. Variations in vertebratetype steroids during testicular maturation in the marine crab, Charybdis natator (Herbst). J Mar Biol Assoc India 48: 241– 244; 2006.
Moriyasu M, Benhalima K, Duggan D et al. Reproductive biology of male Jonah crab, Cancer borealis Stimpson, 1859 (Decapoda, Cancridae) on the Scotian Shelf, Northwestern Atlantic. Crustaceana 75: 891–913; 2002.
Jamieson BGM, Tudge CC. Crustacea- Decapoda. In: Jamieson BGM, Wiley CJ eds. Progress in Male Gamete Ultrastructure and Phylogeny; 2000: Vol. 9 Part C, pp. 1–95.
Tudge CC, Scheltinga DM. Spermatozoa morphology of the freshwater anomuran Aegla longirostri Bond-Buckup & Buckup, 1994 (Crustacea: Decapoda: Aeglidae) from South America. Proc Biol Soc of Washington 115: 118–128, 2002.
Scelzo MA, Medina A, Tudge CC. Spermatozoal ultrastructure of the hermit crab Loxopagurus loxochelis (Moreira, 1901) (Decapoda: Anomura: Diogenidae) from the southwestern Atlantic. In: Asakura A. ed. Biology of Anomura II. Crustacean Research; 2006: Vol. 6, pp. 1-11.
Mouchet S. Mode de formation des spermatophores chez quelques Pagures. Comptes Rendus Hebdomadaire des Séances de l’Academie des Sciences, Paris 190: 691–693; 1930.
Mouchet S. Spermatophores des Crustacés Décapodes Anomoures et Brachyoures et castration parasitare chez quelques Pagures. Annales de la Station Océanographique de salammbô 6: 1–203; 1931.
Hamon M. La constitution chimique des de Custaceés supérieurs du groupe des Pagurides. Comptes Rendus Hebdomadaire des Séances de l’Academie des Sciences, Paris 204: 1504–1506; 1937.
Hamon M. Les mechanisms produisant la dehiscence des spermatophores d’Eupagurus prideauxi Leach. Comptes Rendus Hebdomadaire des Séances de la Société de Biologie 130: 1312–135; 1939.
Hamon M. Researches sur les spermatophores. Théses présentées a las faculté Sciences de l’ Université d’Alger, Maĺson-Carrée, Alger 1–185; 1942.
Mathews DC. The development of the pedunculate spermatophore of a hermit crab, Dardanus asper (DeHann). Pacific Sci 7: 255–266; 1953.
Mathews DC. The origin of the spermatophoric mass of the sand crab, Hippa pacifica. Quart J Microsc Sci 97: 257–268; 19 Mathews DC. Further evidences anomuran non-pedunculate spermatophores. Pacific Sci 11: 380–385; 1957.
Greenwood JG. The male reproductive system and spermatophore formation in Pagurus novae-zealandiae (Dana) (Anomura: Paguridae). J Nat Hist 6: 561–574; 1972.
Spalding JF. The nature and formation of spermatophore and sperm plug in Carcinus maenas. Q J Microsc Sci 83: 399–422; 19 Cronin LE. Anatomy and histology of the male reproductive system of Callinectes sapidus. Rathbun. J Morphol 81: 209–239; 19 Ryan EP. Structure and function of the reproductive system of the crab Portunus sanguinolentus (Herbst) (Brachyura: Portunidae). Mar Biol Assoc India (Sympo. Ser.) 2: 506; 1967.
Ryan EP. Spermatogenesis and male reproductive system in the Hawaiian crab Ranina ranina (L.). In: Engeles W. ed. Advances in Invertebrate Reproduction. Elsevier; 1984: p. 629.
Uma K, Subramoniam T. Histochemical characteristics of spermatophore layers of Scylla serrata (Forskal) (Decapoda: Portunidae). Int J Inver Repr 1: 31–41; 1979.
Hinsch GW. Sperm structure of Oxyrhyncha. Cana J Zoolog 51: 421–426; 1973.
Jeyalectumie C, Subramoniam T. Biochemistry of seminal secretions of the crab Scylla serrata with reference to sperm metabolism and storage in the female. Mol Reprod Develop 30: 44–55; 1991.
Bauer RT. Sperm transfer and storage structures in penaeoid shrimps: A functional and phylogentic perspective. In: Bauer RT, Martin W. eds. Crustacean Sexual Biology. New York Columbia University Press; 1991: pp. 183–207.
Diesel R. Structure and function of the reproductive system of the spider crab Inachus phalangium (Decapoda: Majidae): Observations on sperm transfer, sperm storage and spawning. J Crustacean Biol 9: 266–277; 1989.
Tudge CC, Jamieson BGM, Sandberg L et al. Ultrastructure of mature spermatozoon of the king crab Lithodes maja (Lithodidae, Anomura, decapoda): Further confirmation of a lithodid–pagurid relationship. Invertebr Biol 117: 57–66; 1998. Kumar S, Chaudhury K, Sen P et al. Atomic force microscopy: a powerful tool for high-resolution imaging of spermatozoa. J Nanobiotechno 3: 9–13; 2005.
McElfresh MW, Rudd RE, Balhorn R et al. Probing the properties of cells and cell surfaces with the atomic force microscopy. LDRD Final Report 01-ERI-001 (UCRLTR-202443); 2004.
Franken RD. The clinical significance of sperm-zona pellucida binding. Front Biosci 3: 247–253; 1998.