1 H Magnetic Resonance Spectroscopy of live human sperm

AbstractSTUDY QUESTIONCan1H Magnetic Resonance Spectroscopy (MRS) be used to obtain information about the molecules and metabolites in live human spermatozoa?SUMMARY ANSWERPercoll-based density gradient centrifugation (DGC) followed by a further two washing steps, yielded enough sperm with minimal contamination (<0.01%) from seminal fluid to permit effective MRS which detected significant differences (P< 0.05) in the choline/glycerophosphocholine (GPC), lipid and lactate regions of the1H MRS spectrum between sperm in the pellet and those from the 40%/80% interface.WHAT IS KNOWN ALREADYCurrent methods to examine sperm are either limited in their value (e.g. semen analysis) or are destructive (e.g. immunohistochemistry, sperm DNA testing). A few studies have previously used MRS to examine sperm, but these have either looked at seminal plasma from men with different ejaculate qualities or at the molecules present in pooled samples of lyophilized sperm.STUDY DESIGN, SAMPLES/MATERIALS, METHODSSperm suspended in phosphate buffered saline (PBS) at 37 °C were examined by1H MRS scanning using a1H excitation-sculpting solvent suppression sequence after recovery from fresh ejaculates by one of three different methods: (i) simple centrifugation; (ii) DGC with one wash; or (iii) DGC with two washes. In the case of DGC, sperm were collected both from the pellet ( ‘80%’ sperm) and the 40/80 interface (‘40%’ sperm). Spectrum processing was carried out using custom Matlab scrip...
Source: Molecular Human Reproduction - Category: Molecular Biology Source Type: research