-
Something wrong with this record ?
Seminal fluid and sperm diluent affect sperm metabolism in an insect: Evidence from NAD(P)H and flavin adenine dinucleotide autofluorescence lifetime imaging
C. Massino, C. Wetzker, O. Balvin, T. Bartonicka, J. Kremenova, M. Sasinkova, O. Otti, K. Reinhardt
Language English Country United States
Document type Journal Article
Grant support
KR 1666/4-1
Deutsche Forschungsgemeinschaft
OT 521/4-1
Deutsche Forschungsgemeinschaft
18-08468J
Grantová Agentura České Republiky
PubMed
34486193
DOI
10.1002/jemt.23914
Knihovny.cz E-resources
- MeSH
- Flavin-Adenine Dinucleotide * MeSH
- Microscopy, Fluorescence, Multiphoton MeSH
- Sperm Motility MeSH
- NADP * MeSH
- Spermatozoa drug effects MeSH
- Bedbugs MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
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.
Animal Population Ecology Animal Ecology 1 University of Bayreuth Bayreuth Germany
Department of Botany and Zoology Masaryk University Brno Czech Republic
Light Microscopy Facility CMCB Technische Universität Dresden Dresden Germany
References provided by Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc22002995
- 003
- CZ-PrNML
- 005
- 20250312124410.0
- 007
- ta
- 008
- 220113s2022 xxu f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1002/jemt.23914 $2 doi
- 035 __
- $a (PubMed)34486193
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a xxu
- 100 1_
- $a Massino, Christian $u Applied Zoology, Institute of Zoology, Faculty of Biology, Technische Universität Dresden, Dresden, Germany
- 245 10
- $a Seminal fluid and sperm diluent affect sperm metabolism in an insect: Evidence from NAD(P)H and flavin adenine dinucleotide autofluorescence lifetime imaging / $c C. Massino, C. Wetzker, O. Balvin, T. Bartonicka, J. Kremenova, M. Sasinkova, O. Otti, K. Reinhardt
- 520 9_
- $a 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.
- 650 _2
- $a zvířata $7 D000818
- 650 _2
- $a štěnice $7 D001511
- 650 12
- $a flavinadenindinukleotid $7 D005182
- 650 _2
- $a mužské pohlaví $7 D008297
- 650 _2
- $a mikroskopie fluorescenční multifotonová $7 D036641
- 650 12
- $a NADP $7 D009249
- 650 _2
- $a motilita spermií $7 D013081
- 650 _2
- $a spermie $x účinky léků $7 D013094
- 655 _2
- $a časopisecké články $7 D016428
- 700 1_
- $a Wetzker, Cornelia $u Applied Zoology, Institute of Zoology, Faculty of Biology, Technische Universität Dresden, Dresden, Germany $u Light Microscopy Facility, CMCB, Technische Universität Dresden, Dresden, Germany
- 700 1_
- $a Balvín, Ondřej $u Department of Ecology, Faculty of Environmental Sciences, Czech University of Life Sciences Prague, Prague, Czech Republic $7 xx0329898
- 700 1_
- $a Bartonicka, Tomáš $u Department of Botany and Zoology, Masaryk University, Brno, Czech Republic
- 700 1_
- $a Kremenova, Jana $u Department of Botany and Zoology, Masaryk University, Brno, Czech Republic
- 700 1_
- $a Sasinkova, Markéta $u Department of Ecology, Faculty of Environmental Sciences, Czech University of Life Sciences Prague, Prague, Czech Republic
- 700 1_
- $a Otti, Oliver $u Animal Population Ecology, Animal Ecology I, University of Bayreuth, Bayreuth, Germany
- 700 1_
- $a Reinhardt, Klaus $u Applied Zoology, Institute of Zoology, Faculty of Biology, Technische Universität Dresden, Dresden, Germany
- 773 0_
- $w MED00003348 $t Microscopy research and technique $x 1097-0029 $g Roč. 85, č. 1 (2022), s. 398-411
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/34486193 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y p $z 0
- 990 __
- $a 20220113 $b ABA008
- 991 __
- $a 20250312124417 $b ABA008
- 999 __
- $a ok $b bmc $g 1750698 $s 1154144
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2022 $b 85 $c 1 $d 398-411 $e 20210905 $i 1097-0029 $m Microscopy research and technique $n Microsc Res Tech $x MED00003348
- GRA __
- $a KR 1666/4-1 $p Deutsche Forschungsgemeinschaft
- GRA __
- $a OT 521/4-1 $p Deutsche Forschungsgemeinschaft
- GRA __
- $a 18-08468J $p Grantová Agentura České Republiky
- LZP __
- $a Pubmed-20220113
