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Neurolin Ig domain 2 participates in retinal axon guidance and Ig domains 1 and 3 in fasciculation.

Leppert CA, Diekmann H, Paul C, Laessing U, Marx M, Bastmeyer M, Stuermer CA - J. Cell Biol. (1999)

Bottom Line: Repeated intraocular injections of a mAb against Ig domain 2 disturb the disk-directed growth: axons grow in aberrant routes and fail to reach the optic disk, but remain fasciculated. mAbs against Ig domains 1 and 3 disturb the formation of tight fascicles. mAb against Ig domain 2 significantly increases the incidence of growth cone departure from the disk-oriented fascicle track, while mAbs against Ig domains 1 and 3 do not.This was demonstrated by time-lapse videorecording of labeled growth cones.Thus, Ig domain 2 of neurolin is apparently essential for growth cone guidance towards the disk, presumably by being part of a receptor (or complex) for an axon guidance component.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Konstanz, D-78457 Konstanz, Germany.

ABSTRACT
The optic disk-directed growth of retinal ganglion cell axons is markedly disturbed in the presence of polyclonal antineurolin antibodies, which mildly affect fasciculation (Ott, H., M. Bastmeyer, and C.A.O. Stuermer, 1998. J. Neurosci. 18:3363-3372). New monoclonal antibodies (mAbs) against goldfish neurolin, an immunoglobulin (Ig) superfamily cell adhesion/recognition molecule with five Ig domains, were generated to assign function (guidance versus fasciculation) to specific Ig domains. By their ability or failure to recognize Chinese hamster ovary cells expressing recombinant neurolin with deletions of defined Ig domains, mAbs were identified as being directed against Ig domains 1, 2, or 3, respectively. Repeated intraocular injections of a mAb against Ig domain 2 disturb the disk-directed growth: axons grow in aberrant routes and fail to reach the optic disk, but remain fasciculated. mAbs against Ig domains 1 and 3 disturb the formation of tight fascicles. mAb against Ig domain 2 significantly increases the incidence of growth cone departure from the disk-oriented fascicle track, while mAbs against Ig domains 1 and 3 do not. This was demonstrated by time-lapse videorecording of labeled growth cones. Thus, Ig domain 2 of neurolin is apparently essential for growth cone guidance towards the disk, presumably by being part of a receptor (or complex) for an axon guidance component.

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Schematic representation of neurolin deletion clones  and the binding properties of neurolin mAbs. (A) Wild-type neurolin consists of five Ig domains (Ig1–Ig5, black circles), a transmembrane domain (tm), and a short intracellular domain (carboxy terminus, CH3). (B) mAbs against neurolin were screened  for their ability (+) or failure (−) to bind to CHO cells expressing forms of neurolin (ΔIg1, ΔIg2, etc.) from which either one,  two, or three consecutive Ig domains are deleted (white circles).  mAbs fall into four classes: mAbs of class 1 require the presence  of Ig domain 1; mAbs of class 2, Ig domain 2; mAbs of class 3, Ig  domain 3; and mAbs of class 4, Ig domains 1 and 2 (n, number of  mAbs in this class). mAbs N850, N518, N100, and N984 of classes  1, 2, 3, and 4, respectively, were used in functional assays.
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Figure 1: Schematic representation of neurolin deletion clones and the binding properties of neurolin mAbs. (A) Wild-type neurolin consists of five Ig domains (Ig1–Ig5, black circles), a transmembrane domain (tm), and a short intracellular domain (carboxy terminus, CH3). (B) mAbs against neurolin were screened for their ability (+) or failure (−) to bind to CHO cells expressing forms of neurolin (ΔIg1, ΔIg2, etc.) from which either one, two, or three consecutive Ig domains are deleted (white circles). mAbs fall into four classes: mAbs of class 1 require the presence of Ig domain 1; mAbs of class 2, Ig domain 2; mAbs of class 3, Ig domain 3; and mAbs of class 4, Ig domains 1 and 2 (n, number of mAbs in this class). mAbs N850, N518, N100, and N984 of classes 1, 2, 3, and 4, respectively, were used in functional assays.

Mentions: More recently, the IgSF CAM neurolin (Paschke et al., 1992; Laessing et al., 1994), the teleost homologue of DM-GRASP/SC-1/BEN/ALCAM of birds and mammals (Burns et al., 1991; Tanaka et al., 1991; Pourquié et al., 1992; Bowen et al., 1997), has been discovered to contribute to the disk-directed growth of RGC axons in the goldfish retina (Ott et al., 1998). Neurolin, like its avian and mammalian homologues, consists of five extracellular Ig domains, a transmembrane segment, and a short cytoplasmic domain (see Fig. 1 A). The outcome of in vitro assays, probing for DM-GRASP/SC-1/BEN functions, shows that IgSF CAM promotes axon fasciculation by homo- and/ or    heterophilic binding mechanisms (Burns et al., 1991; Tanaka et al., 1991; DeBernado and Chang, 1996). In the chick retina, DM-GRASP participates in the growth of new growth cones along preexisting axons (Pollerberg and Mack, 1994) but a contribution to the oriented growth of axons by this CAM has not been recognized in these earlier studies. That neurolin contributes to the disk-directed growth of RGC axons was evidenced by intraocular injections of Fab fragments of a polyclonal antineurolin antiserum (neurolin Fabs) into the goldfish eye (Ott et al., 1998).


Neurolin Ig domain 2 participates in retinal axon guidance and Ig domains 1 and 3 in fasciculation.

Leppert CA, Diekmann H, Paul C, Laessing U, Marx M, Bastmeyer M, Stuermer CA - J. Cell Biol. (1999)

Schematic representation of neurolin deletion clones  and the binding properties of neurolin mAbs. (A) Wild-type neurolin consists of five Ig domains (Ig1–Ig5, black circles), a transmembrane domain (tm), and a short intracellular domain (carboxy terminus, CH3). (B) mAbs against neurolin were screened  for their ability (+) or failure (−) to bind to CHO cells expressing forms of neurolin (ΔIg1, ΔIg2, etc.) from which either one,  two, or three consecutive Ig domains are deleted (white circles).  mAbs fall into four classes: mAbs of class 1 require the presence  of Ig domain 1; mAbs of class 2, Ig domain 2; mAbs of class 3, Ig  domain 3; and mAbs of class 4, Ig domains 1 and 2 (n, number of  mAbs in this class). mAbs N850, N518, N100, and N984 of classes  1, 2, 3, and 4, respectively, were used in functional assays.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2132892&req=5

Figure 1: Schematic representation of neurolin deletion clones and the binding properties of neurolin mAbs. (A) Wild-type neurolin consists of five Ig domains (Ig1–Ig5, black circles), a transmembrane domain (tm), and a short intracellular domain (carboxy terminus, CH3). (B) mAbs against neurolin were screened for their ability (+) or failure (−) to bind to CHO cells expressing forms of neurolin (ΔIg1, ΔIg2, etc.) from which either one, two, or three consecutive Ig domains are deleted (white circles). mAbs fall into four classes: mAbs of class 1 require the presence of Ig domain 1; mAbs of class 2, Ig domain 2; mAbs of class 3, Ig domain 3; and mAbs of class 4, Ig domains 1 and 2 (n, number of mAbs in this class). mAbs N850, N518, N100, and N984 of classes 1, 2, 3, and 4, respectively, were used in functional assays.
Mentions: More recently, the IgSF CAM neurolin (Paschke et al., 1992; Laessing et al., 1994), the teleost homologue of DM-GRASP/SC-1/BEN/ALCAM of birds and mammals (Burns et al., 1991; Tanaka et al., 1991; Pourquié et al., 1992; Bowen et al., 1997), has been discovered to contribute to the disk-directed growth of RGC axons in the goldfish retina (Ott et al., 1998). Neurolin, like its avian and mammalian homologues, consists of five extracellular Ig domains, a transmembrane segment, and a short cytoplasmic domain (see Fig. 1 A). The outcome of in vitro assays, probing for DM-GRASP/SC-1/BEN functions, shows that IgSF CAM promotes axon fasciculation by homo- and/ or    heterophilic binding mechanisms (Burns et al., 1991; Tanaka et al., 1991; DeBernado and Chang, 1996). In the chick retina, DM-GRASP participates in the growth of new growth cones along preexisting axons (Pollerberg and Mack, 1994) but a contribution to the oriented growth of axons by this CAM has not been recognized in these earlier studies. That neurolin contributes to the disk-directed growth of RGC axons was evidenced by intraocular injections of Fab fragments of a polyclonal antineurolin antiserum (neurolin Fabs) into the goldfish eye (Ott et al., 1998).

Bottom Line: Repeated intraocular injections of a mAb against Ig domain 2 disturb the disk-directed growth: axons grow in aberrant routes and fail to reach the optic disk, but remain fasciculated. mAbs against Ig domains 1 and 3 disturb the formation of tight fascicles. mAb against Ig domain 2 significantly increases the incidence of growth cone departure from the disk-oriented fascicle track, while mAbs against Ig domains 1 and 3 do not.This was demonstrated by time-lapse videorecording of labeled growth cones.Thus, Ig domain 2 of neurolin is apparently essential for growth cone guidance towards the disk, presumably by being part of a receptor (or complex) for an axon guidance component.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Konstanz, D-78457 Konstanz, Germany.

ABSTRACT
The optic disk-directed growth of retinal ganglion cell axons is markedly disturbed in the presence of polyclonal antineurolin antibodies, which mildly affect fasciculation (Ott, H., M. Bastmeyer, and C.A.O. Stuermer, 1998. J. Neurosci. 18:3363-3372). New monoclonal antibodies (mAbs) against goldfish neurolin, an immunoglobulin (Ig) superfamily cell adhesion/recognition molecule with five Ig domains, were generated to assign function (guidance versus fasciculation) to specific Ig domains. By their ability or failure to recognize Chinese hamster ovary cells expressing recombinant neurolin with deletions of defined Ig domains, mAbs were identified as being directed against Ig domains 1, 2, or 3, respectively. Repeated intraocular injections of a mAb against Ig domain 2 disturb the disk-directed growth: axons grow in aberrant routes and fail to reach the optic disk, but remain fasciculated. mAbs against Ig domains 1 and 3 disturb the formation of tight fascicles. mAb against Ig domain 2 significantly increases the incidence of growth cone departure from the disk-oriented fascicle track, while mAbs against Ig domains 1 and 3 do not. This was demonstrated by time-lapse videorecording of labeled growth cones. Thus, Ig domain 2 of neurolin is apparently essential for growth cone guidance towards the disk, presumably by being part of a receptor (or complex) for an axon guidance component.

Show MeSH
Related in: MedlinePlus