Cryopreservation of mutton snapper ( Lutjanus analis) sperm

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Abstract

This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2) , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10%) and three cooling rates ( -90; -60 and -30°C.min−1) on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g) collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P < 0.05) was achieved by combining extender C ( pH 8.2) with DMSO ( 10%) and cooling rate of -60°C.min−1 ( P < 0.05) . The use of cryopreserved sperm presented fertilization rates higher than 59% validating the present protocol for mutton snapper.

Translated abstract

Este trabalho foi realizado com a finalidade de desenvolver um protocolo de crioconservação do sêmen da cioba Lutjanus analis. Em um experimento fatorial foram analisados os efeitos de três diluentes ( pH 6,1; 7,8 e 8,2) , duas concentrações de dimetilsulfóxido ( DMSO, 5 e 10%) e três velocidades de congelamento ( -90, -60 e -30°C.min−1) sobre a taxa de motilidade e tempo de motilidade espermáticas do sêmen crioconservado. Uma amostra de 30 exemplares com peso médio de 1.261,11 ± 449,0 g, oriunda da natureza, foi mantida em tanques-rede. O sêmen obtido foi congelado empregando-se palhetas criogênicas em vapor de nitrogênio e, posteriormente, transferido para nitrogênio líquido. Posteriormente um teste de fertilização foi realizado para avaliar a viabilidade do sêmen crioconservado. A combinação que propiciou maior taxa de motilidade e tempo de motilidade espermáticas ( P < 0,05) foi proporcionada pelo emprego do diluente de pH 8,2 com 10% de DMSO e uma velocidade de congelamento de -60°C.min−1 ( P < 0,05) . O sêmen crioconservado apresentou taxa de fertilização superior a 59% validando o presente protocolo para a cioba.

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Most cited references 252

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Molecular architecture of the sperm flagella: molecules for motility and signaling.

Kazuo Inaba (2003)

Sperm motility is generated by a highly organized, microtubule-based structure, called the axoneme, which is constructed from approximately 250 proteins. Recent studies have revealed the molecular structures and functions of a number of axonemal components, including the motor molecules, the dyneins, and regulatory substructures, such as radial spoke, central pair, and other accessory structures. The force for flagellar movement is exerted by the sliding of outer-doublet microtubules driven by the molecular motors, the dyneins. Dynein activity is regulated by the radial spoke/central pair apparatus through protein phosphorylation, resulting in flagellar bend propagation. Prior to fertilization, sperm exhibit dramatic motility changes, such as initiation and activation of motility and chemotaxis toward the egg. These changes are triggered by changes in the extracellular ionic environment and substances released from the female reproductive tract or egg. After reception of these extracellular signals by specific ion channels or receptors in the sperm cells, intracellular signals are switched on through tyrosine protein phosphorylation, Ca2+, and cyclic nucleotide-dependent pathways. All these signaling molecules are closely arranged in each sperm flagellum, leading to efficient activation of motility.