{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:07:01Z","timestamp":1785434821386,"version":"3.56.0"},"reference-count":34,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T00:00:00Z","timestamp":1785369600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016834","name":"Chongqing Municipal Health Commission","doi-asserted-by":"publisher","award":["2023FY104"],"award-info":[{"award-number":["2023FY104"]}],"id":[{"id":"10.13039\/100016834","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      The vaginal microbiome is an important component of female reproductive health. Community State Type I (CST-I), which is dominated by\n                      <jats:italic>Lactobacillus crispatus<\/jats:italic>\n                      , is generally considered to be closely associated with a healthy vaginal microecological state. Although the taxonomic composition of CST-I communities remains relatively stable, the transcriptional dynamics of\n                      <jats:italic>Lactobacillus crispatus<\/jats:italic>\n                      across different phases of the menstrual cycle remain unclear.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A healthy reproductive-age woman with a stable CST-I vaginal microbiome was enrolled in this longitudinal study. Vaginal secretion samples were collected at six non-menstrual time points throughout a menstrual cycle. Third-generation full-length 16S rRNA gene sequencing and metatranscriptomic sequencing were performed. Differential transcript expression analysis, KEGG pathway enrichment analysis, and Mfuzz time-series clustering were applied to systematically characterize the transcriptional dynamics of the vaginal microbiome across different phases.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Full-length 16S rRNA gene sequencing confirmed that\n                      <jats:italic>Lactobacillus crispatus<\/jats:italic>\n                      remained the dominant species at all six sampling time points, with no substantial changes in overall community composition. However, metatranscriptomic analysis revealed pronounced phase-specific transcriptional reprogramming. During the pre-ovulatory phase, pathways involved in DNA replication, biosynthesis, and central carbon metabolism were upregulated. On the first day after ovulation, pathways associated with carbohydrate utilization, quorum sensing, and redox homeostasis were selectively upregulated. During the mid-to-late luteal phase, pathways related to cell proliferation and growth were generally downregulated, whereas pathways involved in cofactor biosynthesis, the pentose phosphate pathway, and D-amino acid metabolism showed increased expression. Mfuzz clustering further revealed distinct phase-specific functional transitions in\n                      <jats:italic>L. crispatus<\/jats:italic>\n                      that were absent in non-\n                      <jats:italic>L. crispatus<\/jats:italic>\n                      lactobacilli.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The ecological dominance of\n                      <jats:italic>Lactobacillus crispatus<\/jats:italic>\n                      is maintained not solely through numerical abundance, but also through rhythmic and phase-specific transcriptional regulation. Dynamic functional remodeling and metabolic plasticity may represent important mechanisms underlying its long-term persistence and ecological dominance within healthy CST-I vaginal communities.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0354930","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T17:37:56Z","timestamp":1785433076000},"page":"e0354930","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Longitudinal omics study of transcriptional dynamics in Lactobacillus crispatus"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2127-6941","authenticated-orcid":true,"given":"SiLing","family":"Ren","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huimin","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohua","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daju","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liping","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Lan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liangdan","family":"Tang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,30]]},"reference":[{"key":"pone.0354930.ref001","doi-asserted-by":"crossref","first-page":"107141","DOI":"10.1016\/j.micpath.2024.107141","article-title":"Vaginal colonization of Lactobacilli: Mechanism and function","volume":"198","author":"H Xu","year":"2025","journal-title":"Microb Pathog"},{"key":"pone.0354930.ref002","doi-asserted-by":"crossref","first-page":"4680","DOI":"10.1073\/pnas.1002611107","article-title":"Vaginal microbiome of reproductive-age women","author":"J Ravel","year":"2011","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"132","key":"pone.0354930.ref003","doi-asserted-by":"crossref","DOI":"10.1126\/scitranslmed.3003605","article-title":"Temporal dynamics of the human vaginal microbiota","volume":"4","author":"P Gajer","year":"2012","journal-title":"Sci Transl Med"},{"key":"pone.0354930.ref004","unstructured":"University of Cincinnati College of Medicine. Regulation of the hypothalamic-pituitary-ovarian axis. Reproductive Physiology Online. https:\/\/med.uc.edu\/landing-pages\/reproductivephysiology\/lecture-4\/regulation-of-the-hypothalamic-pituitary-ovarian-axis. Accessed 2023 October 1."},{"issue":"3","key":"pone.0354930.ref005","first-page":"331","article-title":"Unraveling the Dynamics of the Human Vaginal Microbiome","volume":"89","author":"KL Nunn","year":"2016","journal-title":"Yale J Biol Med"},{"issue":"2","key":"pone.0354930.ref006","first-page":"136","article-title":"Morphometric characteristics of the vaginal epithelium during the menstrual cycle","volume":"26","author":"I Sj\u00f6berg","year":"1988","journal-title":"Gynecol Obstet Invest"},{"key":"pone.0354930.ref007","first-page":"289","article-title":"Structure and macromolecular properties of cervical mucus glycoproteins","volume":"43","author":"I Carlstedt","year":"1989","journal-title":"Symp Soc Exp Biol"},{"issue":"11","key":"pone.0354930.ref008","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1007\/BF01950032","article-title":"Antagonistic effect of progesterone towards estradiol dipropionate-induced changes in glycogen content in uterus and vagina of P-mice","volume":"41","author":"G Tripathi","year":"1985","journal-title":"Experientia"},{"issue":"1","key":"pone.0354930.ref009","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1186\/s40168-024-01870-5","article-title":"Defining Vaginal Community Dynamics: daily microbiome transitions, the role of menstruation, bacteriophages, and bacterial genes","volume":"12","author":"LW Hugerth","year":"2024","journal-title":"Microbiome"},{"issue":"1","key":"pone.0354930.ref010","first-page":"12","article-title":"Expert consensus on the standardization of clinical laboratory evaluation and reporting of vaginal microecology","volume":"14","author":"Technology The Clinical Laboratory Science and Technology Transformation Professional Committee of the China Association for Promotion of Health Science and","year":"2026","journal-title":"Chinese Journal of Clinical Laboratory Management (Electronic Edition)"},{"issue":"9","key":"pone.0354930.ref011","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.1097\/CM9.0000000000000211","article-title":"Evaluation of the vaginal microbiome in clinical diagnosis and management of vaginal infectious diseases","volume":"132","author":"T Li","year":"2019","journal-title":"Chin Med J (Engl)"},{"issue":"8","key":"pone.0354930.ref012","doi-asserted-by":"crossref","first-page":"2461","DOI":"10.1128\/AEM.02272-07","article-title":"Critical evaluation of two primers commonly used for amplification of bacterial 16S rRNA genes","volume":"74","author":"JA Frank","year":"2008","journal-title":"Appl Environ Microbiol"},{"issue":"10","key":"pone.0354930.ref013","doi-asserted-by":"crossref","first-page":"1674","DOI":"10.1093\/bioinformatics\/btv033","article-title":"MEGAHIT: an ultra-fast single-node solution for large and complex metagenomics assembly via succinct de Bruijn graph","volume":"31","author":"D Li","year":"2015","journal-title":"Bioinformatics"},{"issue":"3","key":"pone.0354930.ref014","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1101\/gr.5969107","article-title":"MEGAN analysis of metagenomic data","volume":"17","author":"DH Huson","year":"2007","journal-title":"Genome Res"},{"issue":"6","key":"pone.0354930.ref015","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1093\/humupd\/dmv038","article-title":"Menstrual physiology: implications for endometrial pathology and beyond","volume":"21","author":"JA Maybin","year":"2015","journal-title":"Hum Reprod Update"},{"issue":"6","key":"pone.0354930.ref016","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1086\/313818","article-title":"Influence of the normal menstrual cycle on vaginal tissue, discharge, and microflora","volume":"30","author":"DA Eschenbach","year":"2000","journal-title":"Clin Infect Dis"},{"issue":"8","key":"pone.0354930.ref017","doi-asserted-by":"crossref","first-page":"980","DOI":"10.1177\/1933719115570906","article-title":"Characterization of a Hormone-Responsive Organotypic Human Vaginal Tissue Model: Morphologic and Immunologic Effects","volume":"22","author":"S Ayehunie","year":"2015","journal-title":"Reprod Sci"},{"issue":"1","key":"pone.0354930.ref018","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1093\/biolreprod\/7.1.9","article-title":"Glycogen content in the vaginal tissue of normally cycling and estrogen and progesterone-treated rhesus monkeys","volume":"7","author":"AT Gregoire","year":"1972","journal-title":"Biol Reprod"},{"issue":"1","key":"pone.0354930.ref019","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1093\/biolreprod\/6.1.117","article-title":"Differentiation of vaginal epithelium in the normal and hormone-treated rhesus monkey","volume":"6","author":"PF Parakkal","year":"1972","journal-title":"Biol Reprod"},{"issue":"3","key":"pone.0354930.ref020","first-page":"3","article-title":"The discovery of different types of cervical mucus and the Billings ovulation method","volume":"21","author":"E Odeblad","year":"1994","journal-title":"Bull Nat Fam Plann Counc Vic"},{"issue":"20","key":"pone.0354930.ref021","doi-asserted-by":"crossref","DOI":"10.1172\/jci.insight.177899","article-title":"Progesterone promotes CXCl2-dependent vaginal neutrophil killing by activating cervical resident macrophage-neutrophil crosstalk","volume":"9","author":"C G\u00f3mez-Oro","year":"2024","journal-title":"JCI Insight"},{"issue":"1","key":"pone.0354930.ref022","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1007\/s00404-009-1044-x","article-title":"Effects of increasing ratio of progesterone in estrogen\/progesterone combination on total oxidant\/antioxidant status in rat uterus and plasma","volume":"281","author":"A Hekimoglu","year":"2010","journal-title":"Arch Gynecol Obstet"},{"key":"pone.0354930.ref023","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1186\/1477-7827-10-49","article-title":"The effects of oxidative stress on female reproduction: a review","volume":"10","author":"A Agarwal","year":"2012","journal-title":"Reprod Biol Endocrinol"},{"key":"pone.0354930.ref024","doi-asserted-by":"crossref","first-page":"742","DOI":"10.3389\/fmicb.2015.00742","article-title":"NADPH-generating systems in bacteria and archaea","volume":"6","author":"SK Spaans","year":"2015","journal-title":"Front Microbiol"},{"issue":"11","key":"pone.0354930.ref025","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1017\/S0007114514000178","article-title":"Riboflavin (vitamin B\u2082) and oxidative stress: a review","volume":"111","author":"M Ashoori","year":"2014","journal-title":"Br J Nutr"},{"issue":"5","key":"pone.0354930.ref026","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1007\/s00018-010-0571-8","article-title":"Emerging knowledge of regulatory roles of D-amino acids in bacteria","volume":"68","author":"F Cava","year":"2011","journal-title":"Cell Mol Life Sci"},{"key":"pone.0354930.ref027","doi-asserted-by":"crossref","first-page":"1725887","DOI":"10.3389\/fmicb.2026.1725887","article-title":"Exploring the molecular crosstalk between the sex steroids drospirenone and ethinylestradiol with vaginal lactobacilli","volume":"17","author":"A Musc\u00f2","year":"2026","journal-title":"Front Microbiol"},{"issue":"3","key":"pone.0354930.ref028","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1111\/j.1574-695X.2006.00162.x","article-title":"Lactobacillus strains isolated from the vaginal microbiota of healthy women inhibit Prevotella bivia and Gardnerella vaginalis in coculture and cell culture","volume":"48","author":"F Atassi","year":"2006","journal-title":"FEMS Immunol Med Microbiol"},{"key":"pone.0354930.ref029","doi-asserted-by":"crossref","first-page":"102991","DOI":"10.1016\/j.anaerobe.2025.102991","article-title":"Metabolism of Lactobacillus and Gardnerella vaginalis in vaginal defined media","volume":"95","author":"V Horrocks","year":"2025","journal-title":"Anaerobe"},{"key":"pone.0354930.ref030","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1186\/s12866-015-0608-0","article-title":"Lactobacillus crispatus inhibits growth of Gardnerella vaginalis and Neisseria gonorrhoeae on a porcine vaginal mucosa model","volume":"15","author":"LM Breshears","year":"2015","journal-title":"BMC Microbiol"},{"issue":"1","key":"pone.0354930.ref031","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1186\/s12866-023-02916-8","article-title":"Glycogen availability and pH variation in a medium simulating vaginal fluid influence the growth of vaginal Lactobacillus species and Gardnerella vaginalis","volume":"23","author":"S Navarro","year":"2023","journal-title":"BMC Microbiol"},{"issue":"1","key":"pone.0354930.ref032","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1186\/s12864-023-09665-y","article-title":"Comparative pangenomic analysis of predominant human vaginal lactobacilli strains towards population-specific adaptation: understanding the role in sustaining a balanced and healthy vaginal microenvironment","volume":"24","author":"A Bhattacharya","year":"2023","journal-title":"BMC Genomics"},{"key":"pone.0354930.ref033","doi-asserted-by":"crossref","first-page":"100330","DOI":"10.1016\/j.bioflm.2025.100330","article-title":"Characterizing species-specific metabolic signatures in vaginal microbiota across planktonic and biofilm states","volume":"10","author":"S Jena","year":"2025","journal-title":"Biofilm"},{"issue":"1","key":"pone.0354930.ref034","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1186\/s13059-022-02635-9","article-title":"Insight into the ecology of vaginal bacteria 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