Immunohistochemical evidence for an intracellular localization of plasma proteins in human foetal choroid plexus and brain

Møllgård, Kjeld; Jacobsen, Marianne; Gk, Jacobsen; Pp, Clausen; Saunders, NR

doi:10.1016/0304-3940(79)95349-7

Cited by 120 publications

(39 citation statements)

References 10 publications

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“…The selective accumulation into lipid droplets of [3H]-AFP conforms to that particular behaviour. On the other hand, the incorporation of rat serum albumin by rhabdomyosarcoma cells is consistent with previous observations showing that the intracellular presence of serum albumin in the immature central nervous system and other tissues of developing animals follows the same pattern of cell and tissue distribution than AFP (Mollgard et al, 1979;Toran-Allerand, 1980;Trojan & Uriel, 1979, 1982 The latter conforms with a large body of information obtained in the past on the resurgence of foetal patterns of gene expression in cancer (oncofoetal antigens, isoenzymes, etc). (Ibsen & Fishman, 1979;Weinhouse, 1982;Uriel, 1979).…”

Section: Resultssupporting

confidence: 92%

Alphafoetoprotein uptake by cloned cell lines derived from a nickel-induced rat rhabdomyosarcoma

Uriel¹,

Poupon²,

Geuskens³

1983

Br J Cancer

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Summary Rat, mouse, pig and chicken alphafoetoproteins (AFP), rat serum albumin and egg albumin, or their fluoresceinated conjugates were added to cultures of several cloned cell lines isolated from a nickelinduced rat rhabdomyosarcoma. The intracellular uptake of assayed proteins was revealed by the indirect immunoperoxidase technique and/or by direct fluorescence microscopy. All (Trojan & Uriel, 1979;Uriel et al., 1982) and birds (Moro & Uriel, 1981). We have subsequently demonstrated that neuron-like elements in primary cultures of dissociated cells from foetal mouse brain hemispheres can incorporate exogenous AFP . This supports the conclusion that the wide distribution of intracellular AFP through the immature nervous system results from protein uptake, as opposed to an eventual in situ AFP synthesis. The same conclusion can probably be extended to other foetal tissues of ecto-and mesodermal origin where intracellular AFP has also been demonstrated during normal ontogenic development (Basteris, 1979; Dziadek & Adamson, 1978; (ABE et al., 1976).AFP and other proteins Rat, mouse, pig and chicken AFP were isolated as previously described (De Nechaud & Uriel, 1971;Hassoux et al., 1977; Lampreave et al., 1980;Moro & Uriel, 1981 Kummer et al. (1981) for tritiation of monoclonal antibodies. Briefly 2mg of lyophilized AFP dissolved in 1 ml of 0.1 M Na borate buffer pH 8.5, were added to 1 mCi of dried [3H]NSP. The mixture was kept for 36 min at 4°C with stirring. Labelled AFP was separated from unincorporated reactants by Sephadex G-25 chromatography using PBS, pH 7.2, as the eluant. Fractions containing AFP were pooled. Aliquots, 50pl each, were put in small vials and stored at -18°C until use. The specific activity of the preparation was of 29uCi mg-' AFP. High-resolution autoradiography Cloned cell lines were incubated in 1 ml fresh MEM medium to whom either 0.45 or 0.90 pCi of [3H]-AFP were added. After 3 h at 37°C, the cells were washed 3 times with PBS and fixed for 1 h at 4°C with 1.6% glutaraldehyde in 0.1 Sorensen phosphate buffer, pH 7. After several washes in buffer, including overnight, the cells were postfixed in 2% OS04 in the same buffer, for 1 h, at room temperature, scraped from the dishes and pelleted. The pellets were dehydrated in alcohol and embedded in Epon. Ultrathin sections were harvested on forward-coated copper grids and covered with a monogranular layer of Ilford L4 emulsion, using a loop. Autoradiograms were developed in a phenidon-containing developer, after gold latensification (Bouteille, 1976). The sections were finally stained with uranyl acetate and lead citrate. AFP incubation of cultured cellsImmunochemical and immunocytochemical reactions Specific rabbit antisera to rat and mouse AFP were obtained as previously described (De Nechaud & Uriel, 1971;Hassoux et al., 1977). Pure antibodies were isolated from their respective antisera by affinity chromatography on AFP-immunoabsorbents prepared by the procedure of Avrameas ALPHAFOETOPROTEIN UPTAKE BY RAT RHABDOMYOSARCOMA CLONES ...

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Section: Resultssupporting

confidence: 92%

Alphafoetoprotein uptake by cloned cell lines derived from a nickel-induced rat rhabdomyosarcoma

Uriel¹,

Poupon²,

Geuskens³

1983

Br J Cancer

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“…The local production of transferrin, of course, does not preclude the possibility that transferrin is also delivered to the brain from the bloodstream. Indeed, this is suggested by the presence of transferrin and other plasma proteins in the choroid plexus and also in neurons of the developing brain (18)(19)(20). It is conceivable that the development of the blood brain barrier during ontogeny limits access of the circulating transferrin to the brain, requiring the synthesis of transferrin inside the blood brain barrier for normal brain cell activity.…”

Section: Discussionmentioning

confidence: 99%

Transferrin gene expression visualized in oligodendrocytes of the rat brain by using in situ hybridization and immunohistochemistry.

Bloch¹,

Popovici²,

Levìn³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

191

104

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The presence and production of transferrin in the adult rat brain have been investigated using both immunohistochemistry and in situ hybridization in tissue sections. Indirect immunofluorescence with four distinct antisera against rat and human transferrin and one monoclonal antibody against human transferrin demonstrated labeling of the cytoplasm of oligodendrocytes (a category of glial cells) in most parts of the brain, especially in the white matter. In situ hybridization using rat transferrin 32P-labeled cDNA as a probe revealed the presence of transferrin mRNA in glial cells whose appearance, distribution, and organization exactly matched those of the cells decorated with the transferrin antibodies. These results provide evidence that the transferrin gene is expressed in the central nervous system and that transferrin is synthesized by and stored within oligodendrocytes in the adult rat brain. These data suggest that this molecule could have a specific function in nervous system activity.

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“…Recent studies on the immunocytochemi cal localization of AFP and SA in the nervous Hajert-Germond/Trojan/Uriel/Hauw system of developing primates [6,7] rats [8] mice [9] and birds [10] have demonstrated the intracellular presence of both proteins in most neural crest and neural tube derivatives during a transitory period of their maturation pathways. AFP was detected within the ner vous system of chicken embryos aged be tween 48 and 56 h and 15 and 17 days of incu bation [10].…”

Section: Introductionmentioning

confidence: 99%

In vitro Uptake of Exogenous α-Fetoprotein by Chicken Dorsal Root Ganglia

et al. 1983

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Exogenous chicken α-Fetoprotein (AFP) was added to embryonic chick dorsal root ganglia plated on gelatin-coated tissue culture dishes at different stages during the differentiation process and its intracellular uptake demonstrated by immunocytochemical methods. Other embryonic serum proteins were added as a control. Morphologically well-differentiated neurons (ganglion cells) appeared positively labeled for AFP, contrasting with weakly stained other cell types: these included spindle-shaped cells identified as Schwann cells and larger, often closely packed cells, which could be less mature neurons. Fibroblasts were found negative or faintly stained. No AFP was noticeable in cultures grown in the absence of the protein. These results suggest that the presence of AFP in the developing CNS is for the most part, if not entirely, due to protein uptake as opposed to in situ synthesis.

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Immunohistochemical evidence for an intracellular localization of plasma proteins in human foetal choroid plexus and brain

Cited by 120 publications

References 10 publications

Alphafoetoprotein uptake by cloned cell lines derived from a nickel-induced rat rhabdomyosarcoma

Alphafoetoprotein uptake by cloned cell lines derived from a nickel-induced rat rhabdomyosarcoma

Transferrin gene expression visualized in oligodendrocytes of the rat brain by using in situ hybridization and immunohistochemistry.

In vitro Uptake of Exogenous α-Fetoprotein by Chicken Dorsal Root Ganglia

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