Colheita, análise e criopreservação de sêmen de uma espécie modelo de primata neotropical, Sagui-de-Tufo-Branco (Callithrix jacchus) / Collection, analysis and cryopreservation of semen from a model species, the common marmoset (Callithrix jacchus)

AUTOR(ES)
DATA DE PUBLICAÇÃO

2007

RESUMO

The Callithrix jacchus can be used as a model species for other Neotropical primates species, mainly Callithriquids, for reproductive studies. Semen samples were collected with the following objectives: to improve the penile vibrostimulation technique, validate sperm evaluation techniques for field work, analyse the semen characteristics from animals at 2 colonies (Brazil and Germany), use 3 different freezing protocols in common marmosets sperms, evaluate a cytochemical technique for mitochondrial activity demonstration and evaluate DNA damage in frozen-thawed sperms. The staining techniques used in this work were: Eosin-nigrosin, Trypan-blue Giemsa, simple staining for acrosome, FITC-PSA, SpermOscan®, Spermac®, demonstration of cytochrome c oxidase activity in situ by the oxidation of 3,3'-diaminobenzidina (DAB) and the Comet assay. Freezing was done in 100 µL semen with TESt-Yolk clarified medium plus 4% glycerol using 0.25 mL straws. The freezing protocols tested were: 1) straws at equilibrium phase with a 25°C to 4°C curve in 2 hours, 10 minutes at nitrogen vapour and finally into liquid nitrogen; 2) same as Protocol 1 without the equilibrium phase; and 3) straws directly into liquid nitrogen. All attempts resulted in 83,33% ejaculates. The simple staining technique for acrosome was validated and resulted in an obvious differentiation of the intact acrosome. Semen characteristics evaluation showed no difference between the 2 colonies. Post-thaw evaluation showed all protocols had negative effects on motility and plasmatic membrane integrity with a slightly superiority on Protocol 1. There was no difference between protocols in the mitochondrial activity demonstration and Protocol 2 presented the lowest DNA damage.

ASSUNTO(S)

sêmen animal dna criopreservação animal callithrix jacchus semen diaminobenzidine diaminobenzidina cryopreservation callithrix jacchus dna animal semen

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