title: Sperm Chromatin Immaturity Observed in Short Abstinence Ejaculates Affects DNA Integrity and Longevity In Vitro.
abstract: BACKGROUND
  The influence of ejaculatory abstinence (EA) on semen parameters and subsequent reproductive outcome is still debatable; hence understanding the impact of EA on sperm structural and functional integrity may provide a valuable information on predicting successful clinical outcome.
  
  
  OBJECTIVE
  To understand the influence of EA on sperm chromatin maturity, integrity, longevity and global methylation status.
  
  
  METHODS
  This experimental prospective study included 76 ejaculates from 19 healthy volunteers who provided ejaculates after observing 1, 3, 5 and 7 days of abstinence. Sperm chromatin maturity, DNA integrity and global methylation status were assessed in the neat ejaculate. Sperm motility, DNA integrity and longevity were assessed in the processed fraction of the fresh and frozen-thawed ejaculates to determine their association with the length of EA.
  
  
  RESULTS
  Spermatozoa from 1 day ejaculatory abstinence (EA-1) displayed significantly higher level of sperm chromatin immaturity in comparison to EA-3 (P < 0.05) and EA-5 (P < 0.01) whereas; the number of 5-methyl cytosine immunostained spermatozoa did not vary significantly across groups. On the other hand, in vitro incubation of processed ejaculate from EA-1 resulted in approximately 20 and 40 fold increase in the DNA fragmented spermatozoa at the end of 6 and 24h respectively (P < 0.01-0.001).
  
  
  CONCLUSIONS
  Use of short-term EA for therapeutic fertilization would be a clinically valuable strategy to improve the DNA quality. However, use of such spermatozoa after prolonged incubation in vitro should be avoided as it can carry a substantial risk of transmitting DNA fragmentation to the oocytes.
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Shubhashree
authors.initials: S
authors.lastname: Uppangala
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Sherine Eliza
authors.initials: SE
authors.lastname: Mathai
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Sujith Raj
authors.initials: SR
authors.lastname: Salian
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Dayanidhi
authors.initials: D
authors.lastname: Kumar
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Vikram Jeet
authors.initials: VJ
authors.lastname: Singh
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Fiona
authors.initials: F
authors.lastname: D'Souza
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Guruprasad
authors.initials: G
authors.lastname: Kalthur
authors.affiliation: Department of Community Medicine, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Asha
authors.initials: A
authors.lastname: Kamath
authors.affiliation: Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal-576104, India.
authors.forename: Satish Kumar
authors.initials: SK
authors.lastname: Adiga
mesh_terms: D002470:Cell Survival; D002843:Chromatin / Q000378:metabolism*; D015925:Cryopreservation; D004247:DNA / Q000235:genetics / Q000378:metabolism; D053938:DNA Fragmentation; D019175:DNA Methylation; D004542:Ejaculation; D005298:Fertility; D006801:Humans; D008297:Male; D011446:Prospective Studies; D055101:Semen Analysis; D012662:Semen Preservation; D012746:Sexual Abstinence; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology*; D013997:Time Factors
chemical_list: D002843:Chromatin; D004247:DNA