In:
PLOS ONE, Public Library of Science (PLoS), Vol. 18, No. 4 ( 2023-4-6), p. e0283773-
Abstract:
Mouse spermatogenesis, from spermatogonial stem cell proliferation to sperm formation, can be reproduced in vitro by culturing testis tissue masses of neonatal mice. However, it remains to be determined whether this method is also applicable when testis tissues are further divided into tiny fragments, such as segments of the seminiferous tubule (ST), a minimal anatomical unit for spermatogenesis. In this study, we investigated this issue using the testis of an Acrosin- GFP/Histone H3.3-mCherry ( Acr /H3) double-transgenic mouse and monitored the expression of GFP and mCherry as indicators of spermatogenic progression. Initially, we noticed that the cut and isolated stretches of ST shrunk rapidly and conglomerated. We therefore maintained the isolation of STs in two ways: segmental isolation without truncation or embedding in soft agarose. In both cases, GFP expression was observed by fluorescence microscopy. By whole-mount immunochemical staining, meiotic spermatocytes and round and elongating spermatids were identified as Sycp3-, crescent-form GFP-, and mCherry-positive cells, respectively. Although the efficiency was significantly lower than that with tissue mass culture, we clearly showed that spermatogenesis can be induced up to the elongating spermatid stage even when the STs were cut into short segments and cultured in isolation. In addition, we demonstrated that lowered oxygen tension was favorable for spermatogenesis both for meiotic progression and for producing elongating spermatids in isolated STs. Culturing isolated STs rather than tissue masses is advantageous for explicitly assessing the various environmental parameters that influence the progression of spermatogenesis.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0283773
DOI:
10.1371/journal.pone.0283773.g001
DOI:
10.1371/journal.pone.0283773.g002
DOI:
10.1371/journal.pone.0283773.g003
DOI:
10.1371/journal.pone.0283773.g004
DOI:
10.1371/journal.pone.0283773.s001
DOI:
10.1371/journal.pone.0283773.s002
DOI:
10.1371/journal.pone.0283773.s003
DOI:
10.1371/journal.pone.0283773.s004
DOI:
10.1371/journal.pone.0283773.s005
DOI:
10.1371/journal.pone.0283773.s006
DOI:
10.1371/journal.pone.0283773.s007
DOI:
10.1371/journal.pone.0283773.r001
DOI:
10.1371/journal.pone.0283773.r002
DOI:
10.1371/journal.pone.0283773.r003
DOI:
10.1371/journal.pone.0283773.r004
DOI:
10.1371/journal.pone.0283773.r005
DOI:
10.1371/journal.pone.0283773.r006
DOI:
10.1371/journal.pone.0283773.r007
DOI:
10.1371/journal.pone.0283773.r008
DOI:
10.1371/journal.pone.0283773.r009
DOI:
10.1371/journal.pone.0283773.r010
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2023
detail.hit.zdb_id:
2267670-3
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