PMID- 32840006
OWN - NLM
STAT- MEDLINE
DCOM- 20211019
LR  - 20211019
IS  - 1439-0264 (Electronic)
IS  - 0340-2096 (Linking)
VI  - 50
IP  - 1
DP  - 2021 Jan
TI  - In vivo acridine orange human spermatozoa staining-A new perspective for RNA 
      detection and spermatozoa morphology evaluation.
PG  - 102-107
LID - 10.1111/ahe.12606 [doi]
AB  - Significant increases in male infertility and the still unresolved questions on 
      the compatibility and interpretation of current methods in infertility 
      diagnostics call for new protocols. Morphology, genome damage, RNA content and 
      quantity are currently in practice as the major parameters in evaluation of sperm 
      quality. However, results of various methods are not always in mutual 
      concordance. In this study, in vivo acridine orange (AO) staining, which is 
      presently in application in the estimation of genome damage in reticulocytes, was 
      adjusted for spermatozoa staining. Ten men suffering from 
      oligoasthenoteratozoospermia (OAT) and 10 healthy fertile men were analysed using 
      in vivo AO staining. Microscopic analysis was performed by fluorescent and 
      confocal fluorescent microscopy. Our results show that this method preserves 
      spermatozoa membranes, which enables new insight into spermatozoa genome damage, 
      RNA content in residual cytoplasm, damage of neck area with mitochondrion and 
      tail pathology. The introduced method explains the difference between results of 
      sperm DNA fragmentation assay and the globally used AO staining and opens new 
      options for the development of automated systems. In conclusion, the results of 
      our study offer (a) an innovative approach to the analysis of spermatozoa 
      pathology, (b) enable localization and quantification of RNA in residual 
      cytoplasm, (c) a significant contribution to research of aetiology of infertility 
      in men, (d) open new perspectives for the automatization of sperm quality 
      estimation and (e) improve the personalized approach in the selection of in vitro 
      fertilization protocols.
CI  - (c) 2020 Wiley-VCH GmbH.
FAU - Fucic, Aleksandra
AU  - Fucic A
AUID- ORCID: 0000-0003-3642-1037
AD  - Institute for Medical Research and Occupational Health, Zagreb, Croatia.
AD  - Scientific Centre of Excellence for Reproductive and Regenerative Medicine, 
      University of Zagreb School of Medicine, Zagreb, Croatia.
FAU - Maric, Tihana
AU  - Maric T
AUID- ORCID: 0000-0002-6238-9811
AD  - Scientific Centre of Excellence for Reproductive and Regenerative Medicine, 
      University of Zagreb School of Medicine, Zagreb, Croatia.
AD  - Department of Medical biology, University of Zagreb School of Medicine, Zagreb, 
      Croatia.
FAU - Vicic Bockor, Vedrana
AU  - Vicic Bockor V
AD  - Division of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, 
      Croatia.
FAU - Jezek, Davor
AU  - Jezek D
AD  - Scientific Centre of Excellence for Reproductive and Regenerative Medicine, 
      University of Zagreb School of Medicine, Zagreb, Croatia.
AD  - Department of Histology and Embryology, University of Zagreb School of Medicine, 
      Zagreb, Croatia.
LA  - eng
GR  - KK.01.1.1.01.0008/European Union/
PT  - Journal Article
DEP - 20200824
PL  - Germany
TA  - Anat Histol Embryol
JT  - Anatomia, histologia, embryologia
JID - 7704218
RN  - 63231-63-0 (RNA)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*chemistry
MH  - Humans
MH  - Infertility, Male
MH  - Male
MH  - Microscopy, Fluorescence
MH  - RNA/*chemistry/*isolation & purification
MH  - Sperm Motility/physiology
MH  - Spermatozoa/*cytology
MH  - Staining and Labeling
OTO - NOTNLM
OT  - acridine orange
OT  - confocal fluorescent microscopy
OT  - infertility
OT  - morphology
OT  - spermatozoa
EDAT- 2020/08/26 06:00
MHDA- 2021/10/21 06:00
CRDT- 2020/08/26 06:00
PHST- 2020/05/13 00:00 [received]
PHST- 2020/06/23 00:00 [revised]
PHST- 2020/07/30 00:00 [accepted]
PHST- 2020/08/26 06:00 [pubmed]
PHST- 2021/10/21 06:00 [medline]
PHST- 2020/08/26 06:00 [entrez]
AID - 10.1111/ahe.12606 [doi]
PST - ppublish
SO  - Anat Histol Embryol. 2021 Jan;50(1):102-107. doi: 10.1111/ahe.12606. Epub 2020 
      Aug 24.