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Characterization and expression of the laminin gamma3 chain: a novel, non-basement membrane-associated, laminin chain.

Koch M, Olson PF, Albus A, Jin W, Hunter DD, Brunken WJ, Burgeson RE, Champliaud MF - J. Cell Biol. (1999)

Bottom Line: The gamma3 chain is also a prominent element of the apical surface of ciliated epithelial cells of: lung, oviduct, epididymis, ductus deferens, and seminiferous tubules.The distribution of gamma3-containing laminins on the apical surfaces of a variety of epithelial tissues is novel and suggests that they are not found within ultrastructurally defined basement membranes.It seems likely that these apical laminins are important in the morphogenesis and structural stability of the ciliated processes of these cells.

View Article: PubMed Central - PubMed

Affiliation: The Cutaneous Biology Research Center, Massachusetts General Hospital, and the Department of Dermatology, Harvard Medical School, Charlestown, Massachusetts 02129, USA.

ABSTRACT
Laminins are heterotrimeric molecules composed of an alpha, a beta, and a gamma chain; they have broad functional roles in development and in stabilizing epithelial structures. Here, we identified a novel laminin, composed of known alpha and beta chains but containing a novel gamma chain, gamma3. We have cloned gene encoding this chain, LAMC3, which maps to chromosome 9 at q31-34. Protein and cDNA analyses demonstrate that gamma3 contains all the expected domains of a gamma chain, including two consensus glycosylation sites and a putative nidogen-binding site. This suggests that gamma3-containing laminins are likely to exist in a stable matrix. Studies of the tissue distribution of gamma3 chain show that it is broadly expressed in: skin, heart, lung, and the reproductive tracts. In skin, gamma3 protein is seen within the basement membrane of the dermal-epidermal junction at points of nerve penetration. The gamma3 chain is also a prominent element of the apical surface of ciliated epithelial cells of: lung, oviduct, epididymis, ductus deferens, and seminiferous tubules. The distribution of gamma3-containing laminins on the apical surfaces of a variety of epithelial tissues is novel and suggests that they are not found within ultrastructurally defined basement membranes. It seems likely that these apical laminins are important in the morphogenesis and structural stability of the ciliated processes of these cells.

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Laminin γ3 is expressed on the apical surface of  the ciliated epithelium of the bovine and rat fallopian tube. Confocal laser microscope images of  freshly frozen tissue sections of  fallopian tube; in each image, five  optical sections were superimposed. The laminin γ3-reactive  antiserum R16 (A and D, red)  and anti-laminin α2 chain (A and  D, green) were applied to bovine  tissue together; γ3 immunoreactivity is confined to the apical surface of the epithelium, whereas α2  is present in the basement membrane; D is a higher magnification  of one of the folia in A. Application of the γ3-reactive R21 serum  alone (B) also labels the apical  surface of the bovine epithelium,  but in plaque-like structures. R21  pre-immune serum is nonreactive  (C). Simultaneous visualization of  R21 immunoreactivity (E, red)  and α5 immunoreactivity (E,  green) shows that the γ3 is found  on the apical side of the epithelium and in cytoplasmic stores in  the long processes of these columnar tubal cells whereas α5-immunoreactivity is confined to the  basement membrane. Antibodies  purified from the R21 serum with  the carrier protein, His-thioredoxin are negative (F). γ3 shows a  similar distribution in rat fallopian  tube (G and H): R16 reactivity is  present at the apical surface over  the whole of the ampulla (G) and  in a higher power view of the epithelium (H); comparison of the fluorescent image in H with the differential interference contrast image in I demonstrates that γ3 immunoreactivity is associated with the ciliated surface of the epithelium.
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Figure 7: Laminin γ3 is expressed on the apical surface of the ciliated epithelium of the bovine and rat fallopian tube. Confocal laser microscope images of freshly frozen tissue sections of fallopian tube; in each image, five optical sections were superimposed. The laminin γ3-reactive antiserum R16 (A and D, red) and anti-laminin α2 chain (A and D, green) were applied to bovine tissue together; γ3 immunoreactivity is confined to the apical surface of the epithelium, whereas α2 is present in the basement membrane; D is a higher magnification of one of the folia in A. Application of the γ3-reactive R21 serum alone (B) also labels the apical surface of the bovine epithelium, but in plaque-like structures. R21 pre-immune serum is nonreactive (C). Simultaneous visualization of R21 immunoreactivity (E, red) and α5 immunoreactivity (E, green) shows that the γ3 is found on the apical side of the epithelium and in cytoplasmic stores in the long processes of these columnar tubal cells whereas α5-immunoreactivity is confined to the basement membrane. Antibodies purified from the R21 serum with the carrier protein, His-thioredoxin are negative (F). γ3 shows a similar distribution in rat fallopian tube (G and H): R16 reactivity is present at the apical surface over the whole of the ampulla (G) and in a higher power view of the epithelium (H); comparison of the fluorescent image in H with the differential interference contrast image in I demonstrates that γ3 immunoreactivity is associated with the ciliated surface of the epithelium.

Mentions: The Northern analysis indicated that the laminin γ3 chain was most strongly expressed in placenta, testis, lung, liver, spleen, and ovary. Therefore, we examined the localization of γ3 chains within testis, lung, and ovary. The reactivity within the epididymis and the fallopian tube were particularly striking. Thus, the distribution of γ3 in these tissues was extensively studied. In the female reproductive system, the oviduct was strongly reactive. Cryosections of the bovine (Fig. 7, A–F) or rat (Fig. 7, G–I) ampulla reacted for γ3 using R16 (Fig. 7, A, D, and G–I), or R21 (Fig. 7, B and E) showed brilliant immunoreactivity at the apical surfaces of the tubal mucosa. Double immunofluorescent studies performed with laminin γ3 and either laminin α2 (Fig. 7, A and D) or laminin α5 antibodies (Fig. 7 E) demonstrated that both of these α chains are restricted to the basement membranes of the tubal epithelial and the subjacent endothelium whereas γ3 is expressed at the apical surface. The pre-immune serum from rabbit 21 (Fig. 7 C) was negative, as was the reactivity of anti-thioredoxin antibodies purified from the R21 serum by immunoaffinity (Fig. 7 F).


Characterization and expression of the laminin gamma3 chain: a novel, non-basement membrane-associated, laminin chain.

Koch M, Olson PF, Albus A, Jin W, Hunter DD, Brunken WJ, Burgeson RE, Champliaud MF - J. Cell Biol. (1999)

Laminin γ3 is expressed on the apical surface of  the ciliated epithelium of the bovine and rat fallopian tube. Confocal laser microscope images of  freshly frozen tissue sections of  fallopian tube; in each image, five  optical sections were superimposed. The laminin γ3-reactive  antiserum R16 (A and D, red)  and anti-laminin α2 chain (A and  D, green) were applied to bovine  tissue together; γ3 immunoreactivity is confined to the apical surface of the epithelium, whereas α2  is present in the basement membrane; D is a higher magnification  of one of the folia in A. Application of the γ3-reactive R21 serum  alone (B) also labels the apical  surface of the bovine epithelium,  but in plaque-like structures. R21  pre-immune serum is nonreactive  (C). Simultaneous visualization of  R21 immunoreactivity (E, red)  and α5 immunoreactivity (E,  green) shows that the γ3 is found  on the apical side of the epithelium and in cytoplasmic stores in  the long processes of these columnar tubal cells whereas α5-immunoreactivity is confined to the  basement membrane. Antibodies  purified from the R21 serum with  the carrier protein, His-thioredoxin are negative (F). γ3 shows a  similar distribution in rat fallopian  tube (G and H): R16 reactivity is  present at the apical surface over  the whole of the ampulla (G) and  in a higher power view of the epithelium (H); comparison of the fluorescent image in H with the differential interference contrast image in I demonstrates that γ3 immunoreactivity is associated with the ciliated surface of the epithelium.
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Related In: Results  -  Collection

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Figure 7: Laminin γ3 is expressed on the apical surface of the ciliated epithelium of the bovine and rat fallopian tube. Confocal laser microscope images of freshly frozen tissue sections of fallopian tube; in each image, five optical sections were superimposed. The laminin γ3-reactive antiserum R16 (A and D, red) and anti-laminin α2 chain (A and D, green) were applied to bovine tissue together; γ3 immunoreactivity is confined to the apical surface of the epithelium, whereas α2 is present in the basement membrane; D is a higher magnification of one of the folia in A. Application of the γ3-reactive R21 serum alone (B) also labels the apical surface of the bovine epithelium, but in plaque-like structures. R21 pre-immune serum is nonreactive (C). Simultaneous visualization of R21 immunoreactivity (E, red) and α5 immunoreactivity (E, green) shows that the γ3 is found on the apical side of the epithelium and in cytoplasmic stores in the long processes of these columnar tubal cells whereas α5-immunoreactivity is confined to the basement membrane. Antibodies purified from the R21 serum with the carrier protein, His-thioredoxin are negative (F). γ3 shows a similar distribution in rat fallopian tube (G and H): R16 reactivity is present at the apical surface over the whole of the ampulla (G) and in a higher power view of the epithelium (H); comparison of the fluorescent image in H with the differential interference contrast image in I demonstrates that γ3 immunoreactivity is associated with the ciliated surface of the epithelium.
Mentions: The Northern analysis indicated that the laminin γ3 chain was most strongly expressed in placenta, testis, lung, liver, spleen, and ovary. Therefore, we examined the localization of γ3 chains within testis, lung, and ovary. The reactivity within the epididymis and the fallopian tube were particularly striking. Thus, the distribution of γ3 in these tissues was extensively studied. In the female reproductive system, the oviduct was strongly reactive. Cryosections of the bovine (Fig. 7, A–F) or rat (Fig. 7, G–I) ampulla reacted for γ3 using R16 (Fig. 7, A, D, and G–I), or R21 (Fig. 7, B and E) showed brilliant immunoreactivity at the apical surfaces of the tubal mucosa. Double immunofluorescent studies performed with laminin γ3 and either laminin α2 (Fig. 7, A and D) or laminin α5 antibodies (Fig. 7 E) demonstrated that both of these α chains are restricted to the basement membranes of the tubal epithelial and the subjacent endothelium whereas γ3 is expressed at the apical surface. The pre-immune serum from rabbit 21 (Fig. 7 C) was negative, as was the reactivity of anti-thioredoxin antibodies purified from the R21 serum by immunoaffinity (Fig. 7 F).

Bottom Line: The gamma3 chain is also a prominent element of the apical surface of ciliated epithelial cells of: lung, oviduct, epididymis, ductus deferens, and seminiferous tubules.The distribution of gamma3-containing laminins on the apical surfaces of a variety of epithelial tissues is novel and suggests that they are not found within ultrastructurally defined basement membranes.It seems likely that these apical laminins are important in the morphogenesis and structural stability of the ciliated processes of these cells.

View Article: PubMed Central - PubMed

Affiliation: The Cutaneous Biology Research Center, Massachusetts General Hospital, and the Department of Dermatology, Harvard Medical School, Charlestown, Massachusetts 02129, USA.

ABSTRACT
Laminins are heterotrimeric molecules composed of an alpha, a beta, and a gamma chain; they have broad functional roles in development and in stabilizing epithelial structures. Here, we identified a novel laminin, composed of known alpha and beta chains but containing a novel gamma chain, gamma3. We have cloned gene encoding this chain, LAMC3, which maps to chromosome 9 at q31-34. Protein and cDNA analyses demonstrate that gamma3 contains all the expected domains of a gamma chain, including two consensus glycosylation sites and a putative nidogen-binding site. This suggests that gamma3-containing laminins are likely to exist in a stable matrix. Studies of the tissue distribution of gamma3 chain show that it is broadly expressed in: skin, heart, lung, and the reproductive tracts. In skin, gamma3 protein is seen within the basement membrane of the dermal-epidermal junction at points of nerve penetration. The gamma3 chain is also a prominent element of the apical surface of ciliated epithelial cells of: lung, oviduct, epididymis, ductus deferens, and seminiferous tubules. The distribution of gamma3-containing laminins on the apical surfaces of a variety of epithelial tissues is novel and suggests that they are not found within ultrastructurally defined basement membranes. It seems likely that these apical laminins are important in the morphogenesis and structural stability of the ciliated processes of these cells.

Show MeSH