Lectin binding to rat spermatogenic cells: effects of different fixation methods and proteolytic enzyme treatment.
The binding of a panel of eight different fluorescein-conjugated lectins to rat spermatogenic cells was investigated. Particular attention was paid to the effects of different fixation methods and proteolytic enzyme digestion on the staining pattern. Concanavalin A (Con A), wheatgerm agglutinin (WGA), succinylated WGA (s-WGA) and agglutinin from gorse (UEA I) stained the cytoplasm of most germ cells as well as the spermatid acrosome. In contrast, peanut agglutinin (PNA), castor bean agglutinin (RCAI) and soy bean agglutinin (SBA) mainly stained the acrosome. The staining pattern varied depending on the fixation method used. PNA was particularly sensitive to formalin fixation, while SBA, DBA and UEA I showed decreased binding and Con A, WGA, s-WGA and RCA I were insensitive to this type of fixation. Pepsin treatment of the sections before lectin staining caused marked changes in the staining pattern; staining with PNA in formalin-fixed tissue sections was particularly improved but there was also enhanced staining with SBA and horse gram agglutinin (DBA). On the other hand, in Bouin- and particularly in acetone-fixed tissue sections, pepsin treatment decreased the staining with several of the lectins, for example WGA and UEA I.
Malmi
R
R
Department of Pathology, University of Turku, Finland.
Söderström
K O
KO
D000818:Animals; D003208:Concanavalin A / Q000378:metabolism; D005404:Fixatives / Q000494:pharmacology; D006651:Histocytochemistry / Q000379:methods*; D037102:Lectins / Q000378:metabolism*; D008297:Male; D019887:Peanut Agglutinin; D010447:Peptide Hydrolases / Q000494:pharmacology*; D037121:Plant Lectins; D051381:Rats; D030262:Soybean Proteins; D013090:Spermatocytes / Q000187:drug effects / Q000378:metabolism*; D013737:Testis / Q000187:drug effects; D014909:Wheat Germ Agglutinins / Q000378:metabolism; D010835:Phytohemagglutinins
D003208:Concanavalin A; D005404:Fixatives; D037102:Lectins; D019887:Peanut Agglutinin; D010447:Peptide Hydrolases; D037121:Plant Lectins; D030262:Soybean Proteins; D014909:Wheat Germ Agglutinins; C017707:Ricinus communis agglutinin-1; C036850:Ulex europaeus lectins; C036554:dolichos biflorus agglutinin; C028824:soybean lectin; D010835:Phytohemagglutinins