Pre-treatment of cattle sperm and/or oocyte with antibody to lipocalin type prostaglandin D synthase inhibits in vitro fertilization and increases sperm–oocyte binding
“…This phenomenon may be related to cryocapacitation due to access of ions such as Ca 2+ which turns out to be acrosome and hypermotility reaction [14]. Additionally, in vitro incubation of sperm with L-PGDS protein increases its binding to the zona pellucida [29] an action that is possible only after changes in membrane that occur during the capacitation process [38]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…These two latter molecules are involved in the early capacitation [14], because they change the permeability of the plasma membrane [28]. Additionally, L-PGDS is present in the acrosomal membrane of freshly ejaculated spermatozoa and then disappears within the sperm acrosome reaction [25], and the in vitro incubation of sperm with this protein increases their binding capacity to the zona pellucida [29]. …”
BackgroundThe purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen: the heat shock protein 90 alpha (HSP90a), the Niemann-Pick disease type C2 protein (NPC2), and lipocalin-type prostaglandin D synthase (L-PGDS).Six adult boars (each boar was ejaculated three times, 18 in total) were classified by freezability based on the percentage of functionally competent sperm. The male semen with highest freezability (MHF) and the male semen with lowest freezability (MLF) were centrifuged immediately after collection to separate seminal plasma and spermatozoa to make four possible combinations of these two components and to incubate them for 3 h, adjusting the temperature to 17 °C, to freeze them afterwards. The quantification of proteins was performed in two stages: at zero and at 3 h after incubation of the four combinations.ResultsThe spermatozoa × incubation time (IT) interaction only had effect (P < 0.01) on HSP90a levels; this protein increased in seminal plasma, after 3 h of incubation, in larger quantity (P < 0.05) in combinations with MLF spermatozoa. In relation with the NPC2 protein, two isoforms of 16 and 19 kDa were identified. The 19 kDa isoform was affected (P < 0.01) only by the seminal plasma × IT interaction, with superior values (P < 0.01) both at zero and three hours of incubation, in the combinations with MHF seminal plasma; and 16 kDa isoform was affected (P < 0.01) only by the IT with reduction after 3 h of incubation. The levels of L-PGDS was affected (P < 0.01) only by the spermatozoa × IT interaction, which reduced (P < 0.01) in combinations with MLF spermatozoa after 3 h of incubation.ConclusionsIt is possible to consider that the three proteins evaluated were associated with freezability of boar semen due, especially, to the fact that mixtures with MLF spermatozoa showed greater increase levels of the HSP90a protein and reduction of L-PGDS in plasma. In addition, the seminal plasma of MHF had higher concentration of the NPC2 of 19 kDa protein, which was reduced by incubating with MHF spermatozoa.
“…This phenomenon may be related to cryocapacitation due to access of ions such as Ca 2+ which turns out to be acrosome and hypermotility reaction [14]. Additionally, in vitro incubation of sperm with L-PGDS protein increases its binding to the zona pellucida [29] an action that is possible only after changes in membrane that occur during the capacitation process [38]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…These two latter molecules are involved in the early capacitation [14], because they change the permeability of the plasma membrane [28]. Additionally, L-PGDS is present in the acrosomal membrane of freshly ejaculated spermatozoa and then disappears within the sperm acrosome reaction [25], and the in vitro incubation of sperm with this protein increases their binding capacity to the zona pellucida [29]. …”
BackgroundThe purpose of this study was to determine the association of three proteins involved in sperm function on the freezability of porcine semen: the heat shock protein 90 alpha (HSP90a), the Niemann-Pick disease type C2 protein (NPC2), and lipocalin-type prostaglandin D synthase (L-PGDS).Six adult boars (each boar was ejaculated three times, 18 in total) were classified by freezability based on the percentage of functionally competent sperm. The male semen with highest freezability (MHF) and the male semen with lowest freezability (MLF) were centrifuged immediately after collection to separate seminal plasma and spermatozoa to make four possible combinations of these two components and to incubate them for 3 h, adjusting the temperature to 17 °C, to freeze them afterwards. The quantification of proteins was performed in two stages: at zero and at 3 h after incubation of the four combinations.ResultsThe spermatozoa × incubation time (IT) interaction only had effect (P < 0.01) on HSP90a levels; this protein increased in seminal plasma, after 3 h of incubation, in larger quantity (P < 0.05) in combinations with MLF spermatozoa. In relation with the NPC2 protein, two isoforms of 16 and 19 kDa were identified. The 19 kDa isoform was affected (P < 0.01) only by the seminal plasma × IT interaction, with superior values (P < 0.01) both at zero and three hours of incubation, in the combinations with MHF seminal plasma; and 16 kDa isoform was affected (P < 0.01) only by the IT with reduction after 3 h of incubation. The levels of L-PGDS was affected (P < 0.01) only by the spermatozoa × IT interaction, which reduced (P < 0.01) in combinations with MLF spermatozoa after 3 h of incubation.ConclusionsIt is possible to consider that the three proteins evaluated were associated with freezability of boar semen due, especially, to the fact that mixtures with MLF spermatozoa showed greater increase levels of the HSP90a protein and reduction of L-PGDS in plasma. In addition, the seminal plasma of MHF had higher concentration of the NPC2 of 19 kDa protein, which was reduced by incubating with MHF spermatozoa.
“…PTGDS catalyzes conversion of prostaglandin H2 to prostaglandin D2, a major prostaglandin that regulates many bodily functions including sleep, body temperature, hormone release, and odor responses [44]. Ptgds mRNA has been measured in Sertoli cells and germ cells, and in the epididymis [44], where the protein product is an important component in the seminal fluid and may have a role in fertilization [45], [46]. SIL1 is an adenine nucleotide exchange factor for the heat-shock protein 70 member HSPA5/BiP [47], [48].…”
Current human reproductive risk assessment methods rely on semen and serum hormone analyses, which are not easily comparable to the histopathological endpoints and mating studies used in animal testing. Because of these limitations, there is a need to develop universal evaluations that reliably reflect male reproductive function. We hypothesized that toxicant-induced testicular injury can be detected in sperm using mRNA transcripts as indicators of insult. To test this, we exposed adult male Fischer 344 rats to low doses of model testicular toxicants and classically characterized the testicular injury while simultaneously evaluating sperm mRNA transcripts from the same animals. Overall, this study aimed to: 1) identify sperm transcripts altered after exposure to the model testicular toxicant, 2,5-hexanedione (HD) using microarrays; 2) expand on the HD-induced transcript changes in a comprehensive time course experiment using qRT-PCR arrays; and 3) test these injury indicators after exposure to another model testicular toxicant, carbendazim (CBZ). Microarray analysis of HD-treated adult Fischer 344 rats identified 128 altered sperm mRNA transcripts when compared to control using linear models of microarray analysis (q<0.05). All transcript alterations disappeared after 3 months of post-exposure recovery. In the time course experiment, time-dependent alterations were observed for 12 candidate transcripts selected from the microarray data based upon fold change and biological relevance, and 8 of these transcripts remained significantly altered after the 3-month recovery period (p<0.05). In the last experiment, 8 candidate transcripts changed after exposure to CBZ (p<0.05). The two testicular toxicants produced distinct molecular signatures with only 4 overlapping transcripts between them, each occurring in opposite directions. Overall, these results suggest that sperm mRNA transcripts are indicators of low dose toxicant-induced testicular injury in the rat.
“…Integrins are cell surface receptors that mediate cell-cell and cell-extracellular matrix interactions in many cell types. Many molecules have been characterized and suggested to participate in the fertilization process, having integrin as receptors such as CD46, members of the A Desintegrin and A Metalloprotease domains (ADAM) family, secreted prosphoprotein one (SSP1 or OPN) and molecules with RGD peptides sequence [1][2][3][4][5][6]9,11,[18][19][20][21][22]. Inhibition of spermoocyte adhesion/fusion can be performed by monoclonal antibodies, RGD or desintegrin-like peptides and RNAimediated knockdown [16][17][18][19].…”
Fertilization in mammals requires the successful completion of a sequence of steps, starting with the transport of gametes in the reproductive tract and ending with sperm-egg membrane fusion to produce a zygote. Although some integrin subunits are known to be associated with the plasma membrane of some mammalian oocytes and spermatozoa, the presence of α6 integrin on bovine oocytes with intact zona pellucida has not been reported. The present study was undertaken to evaluate the expression of α6 integrin subunit in bovine oocyte and to determine if in vitro binding to the zona pellucida and fertilization were affected by treating oocytes with α6 integrin subunit antibody. The α6 integrin subunit was identified on the bovine oocyte by immunocytochemistry. In vitro fertilization was significantly decreased when in vitro matured bovine oocytes were pre-incubated with α6 integrin subunit antibody at concentration 5 and 20 μg/mL, and spermoocyte binding increased. These studies demonstrated the presence of α6show abstract
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