Studies on different species, including rats, monkeys and humans, have shown the presence of leukocyte differentiation antigens in the spermatozoa. In some case the expression of these molecules is related to a specific functional state of the sperm cell, as was found for the CD 46 antigen, that in humans can be used as a marker of the acrosome reaction. The aim of the present study was to assess wether promoted lipid peroxidation of the spermatozoa induces any variations in their immunoreactivity with ILA 147 antibody that, in bull spermatozoa, recognizes bovine leukocyte antigens. Freshly ejaculated bovine spermatozoa and cryopreserved semen were tested for ILA 147 reactivity by standard immunoperoxidase staining, before and after promoted lipid peroxidation. Staining intensity was assessed in the individual cells using the microdensitometric method to measure integrated optical density (IOD), overcoming the disadvantage of an operator's subjective interpretation of the results. After the lipid peroxidation there was significantly decreased staining intensity in the fresh spermatozoa, but not in the cryopreserved cells. Furthermore, in the preincubation conditions, the cryopreserved spermatozoa had a lower mean I.O.D. value than the fresh sperm, showing that the freezing and thawing processes induced an alteration in the antigen exposure. However the mean immunoreactivity of the cryopreserved cells was not significantly influenced by lipid peroxidation. The absorbance value maps, made following immunoperoxidase staining by the examined antibody, showed that the reaction sites in the fresh and cryopreserved spermatozoa fell mainly within the periacrosomal region. Moreover, after induced lipid peroxidation there were fewer reaction sites in this domain. The present research has confirmed the presence of the examined leukocyte antigenic determinant in the bull spermatozoa, and suggests that promoted lipid peroxidation and the freezing and thawing of spermatozoa can produce membrane damage, leading to reduced ILA 147 antigenic site exposure.

Effect of lipid peroxidation on the immunocytochemical detection of a leukocyte antigenic determinant in fresh and cryopreserved bovine spermatozoa / D. Meggiolaro, F. Porcelli, A. Lange Consiglio, A. Carnevali, P. Crepaldi, L. Molteni, B. Ferrandi. - In: ITALIAN JOURNAL OF ANIMAL SCIENCE. - ISSN 1594-4077. - 2:4(2003), pp. 255-263. [10.4081/ijas.2003.255]

Effect of lipid peroxidation on the immunocytochemical detection of a leukocyte antigenic determinant in fresh and cryopreserved bovine spermatozoa

D. Meggiolaro
;
F. Porcelli
Secondo
;
A. Lange Consiglio;A. Carnevali;P. Crepaldi;L. Molteni
Penultimo
;
B. Ferrandi
Ultimo
2003

Abstract

Studies on different species, including rats, monkeys and humans, have shown the presence of leukocyte differentiation antigens in the spermatozoa. In some case the expression of these molecules is related to a specific functional state of the sperm cell, as was found for the CD 46 antigen, that in humans can be used as a marker of the acrosome reaction. The aim of the present study was to assess wether promoted lipid peroxidation of the spermatozoa induces any variations in their immunoreactivity with ILA 147 antibody that, in bull spermatozoa, recognizes bovine leukocyte antigens. Freshly ejaculated bovine spermatozoa and cryopreserved semen were tested for ILA 147 reactivity by standard immunoperoxidase staining, before and after promoted lipid peroxidation. Staining intensity was assessed in the individual cells using the microdensitometric method to measure integrated optical density (IOD), overcoming the disadvantage of an operator's subjective interpretation of the results. After the lipid peroxidation there was significantly decreased staining intensity in the fresh spermatozoa, but not in the cryopreserved cells. Furthermore, in the preincubation conditions, the cryopreserved spermatozoa had a lower mean I.O.D. value than the fresh sperm, showing that the freezing and thawing processes induced an alteration in the antigen exposure. However the mean immunoreactivity of the cryopreserved cells was not significantly influenced by lipid peroxidation. The absorbance value maps, made following immunoperoxidase staining by the examined antibody, showed that the reaction sites in the fresh and cryopreserved spermatozoa fell mainly within the periacrosomal region. Moreover, after induced lipid peroxidation there were fewer reaction sites in this domain. The present research has confirmed the presence of the examined leukocyte antigenic determinant in the bull spermatozoa, and suggests that promoted lipid peroxidation and the freezing and thawing of spermatozoa can produce membrane damage, leading to reduced ILA 147 antigenic site exposure.
Bovine spermatozoa; Sperm antigens; Lipid peroxidation; Immunocytochemistry
Settore AGR/17 - Zootecnica Generale e Miglioramento Genetico
Settore VET/01 - Anatomia degli Animali Domestici
Article (author)
File in questo prodotto:
File Dimensione Formato  
Meffiolaro_ijas.2003.pdf

accesso aperto

Tipologia: Publisher's version/PDF
Dimensione 325.42 kB
Formato Adobe PDF
325.42 kB Adobe PDF Visualizza/Apri
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/14714
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? 3
social impact