[Cytometry] Sperm cell sorting - affect on acrosomes

Anna Petrunkina ap542 at cam.ac.uk
Tue Feb 24 11:13:22 EST 2009

Dear Nicole,

the effects may vary, depending upon whether you will sort fresh, 
freshly-diluted, or frozen-thawed sperm. Also, if you sort fresh sperm, 
they might be "hidden" effects which will be revealed if you submit sperm 
to freezing-thawing after sorting.

I would not use FITC-PNA as a single discriminator, at least if there are 
no other checks for plasma membrane functional integrity. The binding sites 
for PNA are being lost in course of acrosome reaction; so you may have 1) 
non fluorescent sperm which has intact acrosomes and 2) non-fluorescent 
sperm which has lost acrosome completely and can not bind FITC-PNA as a 
consequence. Again, the status of sample (fresh, liquid-preserved or 
frozen/thawed) will be important.

Further point to consider is that the PNA binding sites can be located 
either on outer acrosomal membrane, inner acrosomal membrane or within 
acrosome, etc, etc, dependent on species, which will require different 
approach for interpreting your data.

If you look for "Maxwell WM" in Pubmed, there will be a lot of papers on 
sperm sorting and effects it produces on functional and physiological state 
of fresh and frozen-thawed semen in different species. According to his 
work, (Mol Reprod Dev. 1997 Mar;46(3):408-18) sorting increased the amount 
of acrosome reacted sperm in comparison to fresh sample.

Hope that helps,

Dr. Anna Petrunkina
Head of Flow Cytometry
Cambridge Institute for Medical Research
Univeristy of Cambridge
Welcome Trust/MRC Building
Hills Road

--On 23 February 2009 13:32 -0800 Nicole Ravida <nravida at sandiegozoo.org> 

> I was wondering how deleterious the affect of sperm cell sorting is on
> acrosomes. Is there much damage to the membrane?  Can you use FITC-PNA to
> sort the flourescing from non-flourescing sperm and capture the
> non-flourescing sperm population?
> Nicole Ravida
> Senior Laboratory Technician
> San Diego Zoo Conservation Research
> Reproductive Physiology Division
> 15600 San Pasqual Valley Road
> Escondido, CA 92027
> (p) 760-291-5438
> (f) 760-291-5428
> We are the Science of Saving Species
> www.sandiegozoo.org/conservation
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