Immunohistochemical Study of Serum Proteins in Normal and Cadmium-Treated Mouse Testis by in vivo CryotechniqueImmunohistochemical Study of Serum Proteins in Normal and Cadmium-Treated Mouse Testis by in vivo Cryotechnique

Objective To investigate time-dependent changes of serum proteins permeability in the normal and cadmium（Cd）-treated mouse testis, reflecting tight junctional （TJ） barriers of Sertoli cells Methods The serum proteins, albumin and immunoglobulin-G（IgG）, in the seminiferous tubules were firstly immobilized by the “in vivo cryotechnique”, in which the dynamic blood circulation was always kept. The cryofixed testicular tissues were then processed for the freeze-substitution method, and embedded in the paraffin wax. Serial sections of 5μm thickness were immunostained by anti-mouse albumin or IgG antibody with peroxidase immunostaining, and also stained with hematoxylin-eosine （HE）for morphological observation. Results In normal seminiferous tubules, albumin immunoreaction products were localized around peritubular myoid cells and among Leydig cells, as well as in blood vessels. They were also localized as arch-like patterns around some spermatogonia in basal compartments. The number of the immunopositive arch structures was different according to developmental stages of the seminiferous cycle, judging from the arrangement of germ cells by HE-staining. The patterns of localization of IgG immunostaining in normal mouse testis were similar to that of albumin. In 24 h after Cd-treatment, some enlarged spaces and vesicular formation in the seminiferous epithelium were observed on the paraffin sections by HE-staining. The albumin or IgGimmunolocalization was seen not only in the basal compartments, but also in the adluminal compartments between Sertoli.cells and germ cells. Conclusion The structural changes of inter-Sertoli TJ barriers in vivo, such as different immunostaining patterns of serum proteins between the normal and Cd-treated mouse seminiferous tubules, could be clearly detected by the “in vivo cryotechnique” with albumin or IgG immunohistochemistry.

Immunohistochemical Study of Serum Proteins in Normal and Cadmium-Treated Mouse Testis by in vivo Cryotechnique

Objective To investigate time-dependent changes of serum proteins permeability inthe normal and cadmium(Cd)-treated mouse testis, reflecting tight junctional (TJ) barriersof Sertoli cellsMethods The serum proteins, albumin and immunoglobulin-G(IgG), in the seminiferoustubules were firstly immobilized by the "in vivo cryotechnique", in which the dynamicblood circulation was always kept. The cryofixed testicular tissues were then processedfor the freeze-substitution method, and embedded in the paraffin wax. Serial sectionsof 5μm thickness were immunostained by anti-mouse albumin or IgG antibody withperoxidase immunostaining, and also stained with hematoxylin-eosine (HE)formorphological observation.Results In normal seminiferous tubules, albumin immunoreaction products werelocalized around peritubular myoid cells and among Leydig cells, as well as in bloodvessels. They were also localized as arch-like patterns around some spermatogonia inbasal compartments. The number of the immunopositive arch structures was differentaccording to developmental stages of the seminiferous cycle, judging from thearrangement of germ cells by HE-staining. The patterns of localization of IgGimmunostaining in normal mouse testis were similar to that of albumin. In 24 h afterCd-treatment, some enlarged spaces and vesicular formation in the seminiferousepithelium were observed on the paraffin sections by HE-staining. The albumin or IgG immunolocalization was seen not only in the basal compartments, but also in theadluminal compartments between Sertolicells and germ cells.Conclusion The structural changes of inter-Sertoli TJ barriers in vivo, such as differentimmunostaining patterns of serum proteins between the normal and Cd-treated mouseseminiferous tubules, could be clearly detected by the "in vivo cryotechnique" withalbumin or IgG immunohistochemistry.