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Here, we show that most promoters in mouse sperm are flanked by well-positioned nucleosomes marked by active histone modifications.
Although sperm generally maintains fertility for 48 h at cold temperatures, in vitro fertilization rates of C57BL/6 mouse sperm are low after 48-h cold storage.
In the conventional IVF protocol at 1G, mouse sperm are preincubated in vitro with 400 µl of IVF medium for 1 2 h in an incubator at 37°C to develop their fertilization potential (capacitation) [16] and then capacitated spermatozoa are transferred into another 400 µl of IVF medium containing oocytes.
A note of caution for researchers who have not worked with sperm before: mouse sperm are extremely fragile cells.
It is noteworthy that several genes (Adams 1b, 2, 3, 4 and 5) that interact to form functional complexes on mature mouse sperm are among the relaxed/positively selected genes with large average ω branch values (Table 1).
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Mouse sperm were collected from 12 24 week old wild type and hyh male mice.
Permeabilization of human and mouse sperm was accomplished as previously described [17].
Mouse sperm were isolated from the cauda epididymis of 10 mature males and thoroughly washed in PBS.
The tACE3 in fresh mouse sperm were brightly stained in the acrosomal cap area in 100% EtOH-fixed sperm similar to SP56 and ACROSIN [11], [12] (Figure 2).
Mouse sperm were collected from cauda epididymides and capacitated in vitro for 2 h in 200 µl drops of TYH medium covered with paraffin oil.
In our experience (Table 1) and in that of others[2] [5], the impaired fertility associated with cryopreserved mouse sperm is dependent on genetic background, with sperm from the C57BL/6 backgrounds being particularly sensitive.
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Justyna Jupowicz-Kozak
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