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@article{1794820,
author = {Massino, Christian and Wetzker, C. and Balvin, Ondrej and Bartonička, Tomáš and Křemenová, Jana and Sasinkova, Markéta and Otti, Oliver and Reinhardt, Klaus},
article_location = {Hoboken},
article_number = {1},
doi = {http://dx.doi.org/10.1002/jemt.23914},
keywords = {bedbug; Cimex lectularius; FLIM; FLIRR; metabolic mapping; multiphoton microscopy; spermatozoa},
language = {eng},
issn = {1059-910X},
journal = {Microscopy Research and Technique},
title = {Seminal fluid and sperm diluent affect sperm metabolism in an insect: Evidence from NAD(P)H and flavin adenine dinucleotide autofluorescence lifetime imaging},
url = {https://doi.org/10.1002/jemt.23914},
volume = {85},
year = {2022}
}
TY - JOUR
ID - 1794820
AU - Massino, Christian - Wetzker, C. - Balvin, Ondrej - Bartonička, Tomáš - Křemenová, Jana - Sasinkova, Markéta - Otti, Oliver - Reinhardt, Klaus
PY - 2022
TI - Seminal fluid and sperm diluent affect sperm metabolism in an insect: Evidence from NAD(P)H and flavin adenine dinucleotide autofluorescence lifetime imaging
JF - Microscopy Research and Technique
VL - 85
IS - 1
SP - 398-411
EP - 398-411
PB - Wiley
SN - 1059910X
KW - bedbug
KW - Cimex lectularius
KW - FLIM
KW - FLIRR
KW - metabolic mapping
KW - multiphoton microscopy
KW - spermatozoa
UR - https://doi.org/10.1002/jemt.23914
N2 - Sperm metabolism is fundamental to sperm motility and male fertility. Its measurement is still in its infancy, and recommendations do not exist as to whether or how to standardize laboratory procedures. Here, using the sperm of an insect, the common bedbug, Cimex lectularius, we demonstrate that standardization of sperm metabolism is required with respect to the artificial sperm storage medium and a natural medium, the seminal fluid. We used fluorescence lifetime imaging microscopy (FLIM) in combination with time-correlated single-photon counting (TCSPC) to quantify sperm metabolism based on the fluorescent properties of autofluorescent coenzymes, NAD(P)H and flavin adenine dinucleotide. Autofluorescence lifetimes (decay times) differ for the free and protein-bound state of the co-enzymes, and their relative contributions to the lifetime signal serve to characterize the metabolic state of cells. We found that artificial storage medium and seminal fluid separately, and additively, affected sperm metabolism. In a medium containing sugars and amino acids (Grace's Insect medium), sperm showed increased glycolysis compared with a commonly used storage medium, phosphate-buffered saline (PBS). Adding seminal fluid to the sperm additionally increased oxidative phosphorylation, likely reflecting increased energy production of sperm during activation. Our study provides a protocol to measure sperm metabolism independently from motility, stresses that protocol standardizations for sperm measurements should be implemented and, for the first time, demonstrates that seminal fluid alters sperm metabolism. Equivalent protocol standardizations should be imposed on metabolic investigations of human sperm samples.
ER -
MASSINO, Christian, C. WETZKER, Ondrej BALVIN, Tomáš BARTONIČKA, Jana KŘEMENOVÁ, Markéta SASINKOVA, Oliver OTTI and Klaus REINHARDT. Seminal fluid and sperm diluent affect sperm metabolism in an insect: Evidence from NAD(P)H and flavin adenine dinucleotide autofluorescence lifetime imaging. \textit{Microscopy Research and Technique}. Hoboken: Wiley, 2022, vol.~85, No~1, p.~398-411. ISSN~1059-910X. Available from: https://dx.doi.org/10.1002/jemt.23914.
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