Lectin affinity of the seminiferous epithelium in healthy and cryptorchid post‐pubertal boars

Pinart, E.; Bonet, Sergi; Briz, M.; Pastor, Luis Miguel; Sancho, S.; García, Nuria Palacios; Badía, E.; Bassols, Judit

doi:10.1046/j.1365-2605.2001.00282.x

Cited by 23 publications

(18 citation statements)

References 39 publications

(128 reference statements)

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“…As the primitive cells differentiated, DBA staining was lost, which explains why the spermatogonia showed no affinity at all. Similarly, DBA does not bind to spermatogonia in postpubertal boar testes [34]. UCHL1 expression was restricted to germ cells, as reported previously [32].…”

Section: Histochemistrysupporting

confidence: 80%

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Identification, Isolation, and In Vitro Culture of Porcine Gonocytes1

Goel¹,

Sugimoto²,

Minami³

et al. 2007

Biology of Reproduction

117

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Gonocytes are primitive germ cells that reside in the seminiferous tubules of neonatal testes and give rise to spermatogonia, thereby initiating spermatogenesis. Due to a lack of specific markers, the isolation and culture of these cells has proven to be difficult in the pig. In the present study, we show that a lectin, Dolichos biflorus agglutinin (DBA), which has specific affinity for primordial germ cells (PCGs) in the genital ridge, binds specifically to gonocytes in neonatal pig testes. The specific affinity of DBA for germ cells was progressively lost with age. This suggests that DBA binds strongly to primitive germ cells, such as gonocytes, weakly to primitive spermatogonia, and not at all to spermatogonia. The presence of alkaline phosphatase (AP) activity in the germ cells of neonatal pig testis confirmed the existence of primitive germ cells. Gonocytes from neonatal pig testis were purified, and a cell population that consisted of approximately 70% gonocytes was obtained, as indicated by the DBA binding assay. Purified gonocytes were cultured in DMEM/F12 supplemented with 10% FBS in the absence of any specific growth factors for 7 days. The cells remained viable and proliferated actively in culture. Initially, the gonocytes grew as focal colonies that transformed to three-dimensional colonies by 7 days of culture. Cultured germ cells expressed SSEA-1, a marker for embryonic stem (ES) cells, and were negative for the expression of somatic cell markers. These results should help to establish a male germ cell line that could be used for studying spermatogenesis in vitro and for genetic modification of pigs.

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Section: Histochemistrysupporting

confidence: 80%

“…The anti-SSEA-1 and EMA-1 monoclonal antibodies and DBA bind specifically to the large round PGCs of the genital ridge in pigs [33]. In the postpubertal boar testis, DBA binds weakly to the apical cytoplasm of Sertoli cells [34]. In neonatal human testis, DBA binds to spermatogonia and Leydig cells [35].…”

Section: Introductionmentioning

confidence: 99%

Identification, Isolation, and In Vitro Culture of Porcine Gonocytes1

Goel¹,

Sugimoto²,

Minami³

et al. 2007

Biology of Reproduction

117

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“…SJA has specificity toward carbohydrate structures terminating with N-acetylgalactosamine and galactose residues. Finally, VVA recognizes preferentially a-or b-linked terminal Nacetylgalactosamine (Pinart et al, 2001;Nagano et al, 2005). All lectins of the third group generally stained testis cells weakly (note that the different intensities in Fig.…”

Section: Histological Analysismentioning

confidence: 86%

Lectin‐binding pattern as tool to identify and enrich specific primary testis cells of the tilapia (Oreochromis niloticus) and medaka (Oryzias latipes)

Tokalov

Gutzeit

2006

J Exp Zool Pt B

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Cell type-specific lectin binding is a useful tool for the analysis of developing systems. We describe the binding pattern of 21 different fluorescein isothiocyanate (FITC)-labelled lectins to the testis of two model teleost species, the medaka (Oryzias latipes) and the tilapia (Oreochromis niloticus). The analysis of the binding pattern was carried out on tissue sections (medaka and tilapia) and using primary culture cells (only tilapia). Lectin binding was studied by confocal microscopy and for histological analysis some sections were, in addition, stained with bodipy to gain additional information concerning the cytological organization of the cystic mode of spermatogenesis in fish. The observed differences in lectin staining of different cell types in primary cultures were quantified by flow cytometry. Only few lectins bound specifically to haploid cells while the reaction to diploid or tetraploid cells was generally stronger. However, the extracellular material around the haploid spermatids and spermatozoa in spermatocysts showed a strong staining reaction with several lectins (e.g., Phaseolus vulgaris Erythro agglutinin). The apparent differences in the cellular lectin-binding pattern can be used to identify particular cell types, to monitor their differentiation in vitro or to enrich particular cell types from heterogeneous cultures using magnetic beads coated with anti-FITC antibodies. Using the latter approach, we show that it is possible to enrich for gonial cells and at the same time deplete the preparation for haploid cells and Sertoli cells.

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“…In our study, we found very few DBA-positive cells in porcine testis, which is in agreement with Goel et al (2007), who also found a small number of DBA-positive cells in testis of a 5-month-old pig. Pinart et al (2001) reported no DBA affinity to cells of the seminiferous epithelium in 9-month-old postpubertal porcine testes, indicating that in older boars the epitope bound by DBA is no longer present in spermatogonia. Very few cells bound DBA in the horse testis (Ha et al 2003).…”

Section: Discussionmentioning

confidence: 98%

The Sda/GM2-glycan is a carbohydrate marker of porcine primordial germ cells and of a subpopulation of spermatogonia in cattle, pigs, horses and llama

Klisch

Contreras²,

Sun³

et al. 2011

Reproduction

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Spermatogonia are a potential source of adult pluripotent stem cells and can be used for testis germ cell transplantation. Markers for the isolation of these cells are of great importance for biomedical applications. Primordial germ cells and prepubertal spermatogonia in many species can be identified by their binding of Dolichos biflorus agglutinin (DBA). This lectin binds to two different types of glycans, which are a-linked N-acetylgalactosamine (GalNac) and b-linked GalNac, if this is part of the Sda or GM2 glycotopes. We used the MAB CT1, which is specific for the trisaccharides motif NeuAca2-3(GalNAcb1-4)Galb1-, which is common to both Sda and GM2 glycotopes, to further define the glycosylation of DBA binding germ cells. In porcine embryos, CT1 bound to migratory germ cells and gonocytes. CT1/DBA double staining showed that the mesonephros was CT1 negative but contained DBA-positive cells. Gonocytes in the female gonad became CT1 negative, while male gonocytes remained CT1 positive. In immunohistological double staining of cattle, pig, horse and llama testis, DBA and CT1 staining was generally colocalised in a subpopulation of spermatogonia. These spermatogonia were mainly single, sometimes paired or formed chains of up to four cells. Our data show that the Sda/GM2 glycotope is present in developing germ cells and spermatogonia in several species. Owing to the narrower specificity of the CT1 antibody, compared with DBA, the former is likely to be a useful tool for labelling and isolation of these cells.

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Lectin affinity of the seminiferous epithelium in healthy and cryptorchid post‐pubertal boars

Cited by 23 publications

References 39 publications

Identification, Isolation, and In Vitro Culture of Porcine Gonocytes1

Identification, Isolation, and In Vitro Culture of Porcine Gonocytes1

Lectin‐binding pattern as tool to identify and enrich specific primary testis cells of the tilapia (Oreochromis niloticus) and medaka (Oryzias latipes)

The Sda/GM2-glycan is a carbohydrate marker of porcine primordial germ cells and of a subpopulation of spermatogonia in cattle, pigs, horses and llama

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