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Universidade Federal do Rio Grande do Sul – UFRGS




       PROTEÍNAS DO PLASMA
         SEMINAL EQUINO


Disciplina: Tecnologia de Sêmen
Pós graduando: Murilo Farias Rodrigues
Introdução
• As metodologias de avaliação convencional poderão
 identificar garanhões subférteis, porém apresentam uma
 grande variação dos resultados in vivo de garanhões
 considerados férteis (Giesecke et al., 2010)

• Proteínas do plasma seminal modulam funções cruciais
 na reprodução:
 • Motilidade
 • Capacitação espermática
 • Proteção celular
 • Reação acrossômica
 • Fertilização
                                         Moura et al., 2010
Proteínas do Plasma Seminal - SP

• Três famílias principais:
• Proteínas secretadas ricas em cisteínas (CRISPs)
• Proteínas que contém o domínio fibronectina tipo II
• Proteínas da família espermadesinas




                                        Töpfer-Petersen et al., 2005
Adaptado Pousadas, I. C (2007)
Proteínas PS equinos - Estrutura




Töpfer-Petersen et al., 2005
HSP-1 e 2
    • Produzidas na cauda do
      epidídimo
    • Ligam-se a fosfolipídios
      da membrana e a
      heparina



    (Calvete et al., 1999)



Figura fonte: Manjunath e Therien, 2002 - Journal of Reproductive
Immunology 53 (2002) 109–119
CRISP-1

• Liga-se na região
 equatorial durante a
 capacitação
  • Evita a capacitação
   prematura (Roberts, 2006)
CRISP-2
• Produzida no testículo – maturação acrossomática


• TPX-1 – Ativação dos canais de Ca++ (Motilidade)




GIBBS et al. JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 281, NO. 7, pp. 4156–4163, February 17, 2006
CRISP-3 (HSP-3)

• Encontra-se em maior quantidade no plasma seminal da
 espécie equina (Schambony et al., 1998)

• Efeito protetor seletivo contra a ligação celular dos PMN
 (Troedsson et al., 2010)

• Apresenta r=0,495 (P=0,027) com a taxa concepção/1º
 ciclo (Novak et al., 2010)
HSP-4 e HSP-5

• HSP-4:
  • Está relacionado ao produto semelhante ao gene da calcitonina e
    os níveis de calcitonina foram correlacionados com a motilidade
    espermática (Mungan et al., 2001)


• HSP-5:
    • Ainda não se observou relação com as outras proteínas
HSP-6 e HSP-8

• São diferentes isoformas de uma mesma proteínas
 pertencentes à família das calicreínas
 • Taxa de concepção/1ºCiclo com r= - 0,60 (P=0,028)
 • Volume ejaculado com r= 0,46 (P=0,04)




                                             Novak et al., 2010
HSP-7

• Espermadesinas
  • Estabilização da membrana plasmática, capacitação e interação
    espermatozoide-oviduto ou zona pelúcida
                                            (Martinez et al., 2011)




                               Töpfer-Petersen et al., 2005
Osteopontina




       Souza, 2008.
PG- sintase tipo lipocalina (PDGS)

• Papel no desenvolvimento e maturação espermática
 (Gerena et al., 2000).

• Entretanto a exata função que a mesma exerce sobre os
 espermatozóides ainda não está esclarecida (Moura et
 al., 2006).
Outras Proteínas: SPTZ
            Proteína            Fertilidade   Peso molecular (kDa)
                                    (r)
Citrato Sintase                     +                 52
Fumarato Hidratase                  +                 50,9
Malato Desidrogenase                +                 36
Dihidrolipamida Desidrogenase       +                 54
α-Enolase                           +                 41




                                               Adaptado Novak et al., 2010
Outras Proteínas: Plasma Seminal
             Proteína   Fertilidade   Peso molecular (kDa)
                            (r)
CRISP-3                     +                 29,3
Calicreína – IE2             -                29
Clusterina                   -                29
SP2                          -                52
SP1                          -                14,2




                                       Adaptado Novak et al., 2010
IGF-1




  P= 0,007
             Novak et al., 2010
Clusterina




  P= 0,0003
              Novak et al., 2010
Outras Proteínas r = [Enzimática]




                           Pesch, et al. 2006
OBRIGADO

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Proteínas do plasma seminal murilo

  • 1. Universidade Federal do Rio Grande do Sul – UFRGS PROTEÍNAS DO PLASMA SEMINAL EQUINO Disciplina: Tecnologia de Sêmen Pós graduando: Murilo Farias Rodrigues
  • 2. Introdução • As metodologias de avaliação convencional poderão identificar garanhões subférteis, porém apresentam uma grande variação dos resultados in vivo de garanhões considerados férteis (Giesecke et al., 2010) • Proteínas do plasma seminal modulam funções cruciais na reprodução: • Motilidade • Capacitação espermática • Proteção celular • Reação acrossômica • Fertilização Moura et al., 2010
  • 3. Proteínas do Plasma Seminal - SP • Três famílias principais: • Proteínas secretadas ricas em cisteínas (CRISPs) • Proteínas que contém o domínio fibronectina tipo II • Proteínas da família espermadesinas Töpfer-Petersen et al., 2005
  • 5. Proteínas PS equinos - Estrutura Töpfer-Petersen et al., 2005
  • 6. HSP-1 e 2 • Produzidas na cauda do epidídimo • Ligam-se a fosfolipídios da membrana e a heparina (Calvete et al., 1999) Figura fonte: Manjunath e Therien, 2002 - Journal of Reproductive Immunology 53 (2002) 109–119
  • 7. CRISP-1 • Liga-se na região equatorial durante a capacitação • Evita a capacitação prematura (Roberts, 2006)
  • 8. CRISP-2 • Produzida no testículo – maturação acrossomática • TPX-1 – Ativação dos canais de Ca++ (Motilidade) GIBBS et al. JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 281, NO. 7, pp. 4156–4163, February 17, 2006
  • 9. CRISP-3 (HSP-3) • Encontra-se em maior quantidade no plasma seminal da espécie equina (Schambony et al., 1998) • Efeito protetor seletivo contra a ligação celular dos PMN (Troedsson et al., 2010) • Apresenta r=0,495 (P=0,027) com a taxa concepção/1º ciclo (Novak et al., 2010)
  • 10. HSP-4 e HSP-5 • HSP-4: • Está relacionado ao produto semelhante ao gene da calcitonina e os níveis de calcitonina foram correlacionados com a motilidade espermática (Mungan et al., 2001) • HSP-5: • Ainda não se observou relação com as outras proteínas
  • 11. HSP-6 e HSP-8 • São diferentes isoformas de uma mesma proteínas pertencentes à família das calicreínas • Taxa de concepção/1ºCiclo com r= - 0,60 (P=0,028) • Volume ejaculado com r= 0,46 (P=0,04) Novak et al., 2010
  • 12. HSP-7 • Espermadesinas • Estabilização da membrana plasmática, capacitação e interação espermatozoide-oviduto ou zona pelúcida (Martinez et al., 2011) Töpfer-Petersen et al., 2005
  • 13. Osteopontina Souza, 2008.
  • 14. PG- sintase tipo lipocalina (PDGS) • Papel no desenvolvimento e maturação espermática (Gerena et al., 2000). • Entretanto a exata função que a mesma exerce sobre os espermatozóides ainda não está esclarecida (Moura et al., 2006).
  • 15. Outras Proteínas: SPTZ Proteína Fertilidade Peso molecular (kDa) (r) Citrato Sintase + 52 Fumarato Hidratase + 50,9 Malato Desidrogenase + 36 Dihidrolipamida Desidrogenase + 54 α-Enolase + 41 Adaptado Novak et al., 2010
  • 16. Outras Proteínas: Plasma Seminal Proteína Fertilidade Peso molecular (kDa) (r) CRISP-3 + 29,3 Calicreína – IE2 - 29 Clusterina - 29 SP2 - 52 SP1 - 14,2 Adaptado Novak et al., 2010
  • 17. IGF-1 P= 0,007 Novak et al., 2010
  • 18. Clusterina P= 0,0003 Novak et al., 2010
  • 19. Outras Proteínas r = [Enzimática] Pesch, et al. 2006