Proteomic profiling of the signal crayfish Pacifastacus leniusculus egg and spermatophore

Abstract

Proteins of the signal crayfish Pacifastacus leniusculus egg and spermatophore were identified using in-gel digestion, mass spectrometry, and Mascot search. Forty-one and one-hundred-fifty proteins were identified in egg and spermatophore, respectively. The proteins were classified into nine categories including cell defence, cell signaling, cytoskeleton, DNA related activity, metabolism and energy production, protease and protease inhibitor, respiration, transportation, and others and unknown. Twenty-two proteins were found in both egg and spermatophore. The respiration and cytoskeleton groups are the most diverse categories in the protein profiles of the egg and spermatophore, respectively. No protein was assigned to DNA related activity and cell defence categories in the protein profile of the crayfish egg. Differences between protein profiles of the crayfish egg and spermatophore show different functional priorities for each of gametes. Several proteins having possible roles in gametogenesis, capacitation, acrosome reaction, and fertilization were identified. This proteomic profile of signal crayfish gametes provides a basis for further investigation of functional roles of the identified proteins in aspects of reproduction such as capacitation and fertilization.

Fine structure of the spermatozoon in three species of Cambaridae (Arthropoda: Crustacea: Decapoda)Cambarus robustus,Orconectes propinquusandOrconectes rusticus: a comparative biometrical study

“…The radial arms in decapod spermatozoa may be composed of microtubules, nuclear material, or both (Tudge, 2009). Molecular studies identified tubulin proteins, as major units of microtubules in the proteomic profile of the crayfish male gamete that confirms the microtubular nature of radial arms (Niksirat et al, 2014a; Niksirat et al, 2015b) and as seen in the TEM images in this study. Although, microtubular radial arms undergo protein tyrosine phosphorylation during spermatophore post-mating storage on the body surface of female crayfish (Niksirat et al, in press), the exact role(s) of radial arms in fertilization is yet to be determined.…”

“…A positive staining of Janus green B, an indicator of active mitochondria, in the same area of the crayfish spermatozoon has been reported (André, 1962). Several proteins related to metabolism and energy production were identified in the protein profile of the crayfish male gamete that may confirm the presence of an energy supply center in the sperm cell (Niksirat et al, 2014a; Niksirat et al, 2015b). …”

Label-free protein quantification in freshly ejaculated versus post-mating spermatophores of the noble crayfish Astacus astacus

“…Electrical stimulation was used for extrusion and collection of freshly ejaculated spermatophores from seven males [21,22] (p< 0.01) were only used for the interpretation and discussion. Two experimental groups were analyzed using t-test.…”

Comparative ultrastructure of spermatozoa of the redclaw Cherax quadricarinatus and the yabby Cherax destructor (Decapoda, Parastacidae)

“…Increasing knowledge of final spermatozoon maturation among crayfish families requires study of morphological changes after mating and during fertilization in Cambaridae and Parastacidae (Niksirat et al, 2014b). Related assessment of the proteomic profile of spermatophores (Niksirat et al, 2014a) and morphological changes during egg activation (Niksirat et al, 2015) may provide insight into biological pathways related to capacitation and fertilization in the crayfish. More complete understanding of crayfish reproduction can support development of new techniques for artificial fertilization, facilitating progress in intensive aquaculture and programs for conservation of critically endangered species.…”