Binding of fluorescein isothiocyanate conjugated lectins to rat spermatogenic cells in tissue sections |
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Authors: | K O Söderström R Malmi K Karjalainen |
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Institution: | (1) Departments of Pathology and Anatomy, University of Turku, SF-20520 Turku 52, Finland |
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Abstract: | Summary The testes from three months old Spague-Dawley rats were fixed in Bouin's fluid or neutral buffered 10% formalin, embedded
in paraffin, sectioned and after deparaffination stained with the following fluorescein isothiocyanate coupled lectins: PNA,
WGA, Con A, RCA, SBA, DBA and UEA. The results show that there are considerable differences in the staining pattern of various
spermatogenic cells between different lectins. The fixation in Bouin's fluid enhanced the staining of all the lectins compared
to formalin fixation in which only a weak staining could be seen in the acrosomes of spermatids after WGA or PNA staining.
PNA and WGA stained specifically the acrosome of the developing spermatids, which was seen from the beginning of the acrosome
formation and lasted up to late spermiogenesis. However, the staining with PNA decreased in the late spermatids whereas the
intensity of the staining remained unchanged with WGA. Con A did not stain the acrosome but stained unspecifically the cytoplasm
of all spermatogenic cells. RCA stained faintly the acrosome throughout the spermatid differentiation. DBA and UEA stained
specifically the chromosomes of B spermatogonia. DBA also faintly stained the cell membranes of early spermatids. SBA did
not show any specific staining of the spermatogenic cells. Based on this it is suggested that the carbohydrates and glycoproteins
which are known to be present in the acrosome are formed already in the beginning of the acrosome formation. The decrease
in the PNA staining in late spermatids possibly reflects the fact that the receptor molecules are not synthesized in late
spermatids but are formed in earlier developmental stages and are thereafter preserved in the acrosome. The enhancement of
lectin binding caused by Bouin's fixative might also be applied to other tissues where previous experiments with formalin
fixed tissue have failed to show any staining. |
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