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A novel member of the netrin family, beta-netrin, shares homology with the beta chain of laminin: identification, expression, and functional characterization.

Koch M, Murrell JR, Hunter DD, Olson PF, Jin W, Keene DR, Brunken WJ, Burgeson RE - J. Cell Biol. (2000)

Bottom Line: In addition, beta-netrin is expressed in a limited set of fiber tracts within the brain, including the lateral olfactory tract and the vomeronasal nerve.Functional studies were performed and show that beta-netrin promotes neurite elongation from olfactory bulb explants.Together, these data suggest that beta-netrin is important in neural, kidney, and vascular development.

View Article: PubMed Central - PubMed

Affiliation: Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA.

ABSTRACT
The netrins are a family of laminin-related molecules. Here, we characterize a new member of the family, beta-netrin. beta-Netrin is homologous to the NH(2) terminus of laminin chain short arms; it contains a laminin-like domain VI and 3.5 laminin EGF repeats and a netrin C domain. Unlike other netrins, this new netrin is more related to the laminin beta chains, thus, its name beta-netrin. An initial analysis of the tissue distribution revealed that kidney, heart, ovary, retina, and the olfactory bulb were tissues of high expression. We have expressed the molecule in a eukaryotic cell expression system and made antibodies to the expressed product. Both in situ hybridization and immunohistochemistry were used to describe the cellular source of beta-netrin and where beta-netrin is deposited. beta-Netrin is a basement membrane component; it is present in the basement membranes of the vasculature, kidney, and ovaries. In addition, beta-netrin is expressed in a limited set of fiber tracts within the brain, including the lateral olfactory tract and the vomeronasal nerve. Functional studies were performed and show that beta-netrin promotes neurite elongation from olfactory bulb explants. Together, these data suggest that beta-netrin is important in neural, kidney, and vascular development.

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β-Netrin RNA is expressed widely in human and mouse tissues. The same cDNA probe was used to probe adult human tissue blots (A) and a dot blot (B). Only a single major RNA species is detected on the tissue blots. Strong signals are present in spleen, prostate, ovary, heart, kidney and pancreas, mammary gland, and uterus. Only weak signals are present in brain tissue. (C) RT-PCR confirms strong expression of β-netrin RNA in adult mouse kidney, heart, and ovary, as expected from the human blot analyses. In addition, β-netrin RNA is readily detected in several CNS structures including, olfactory bulb and retina. Some expression is observed in other brain regions; however, spinal cord, brain stem, and dorsal root ganglia contain only low levels of β-netrin RNA.
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Figure 5: β-Netrin RNA is expressed widely in human and mouse tissues. The same cDNA probe was used to probe adult human tissue blots (A) and a dot blot (B). Only a single major RNA species is detected on the tissue blots. Strong signals are present in spleen, prostate, ovary, heart, kidney and pancreas, mammary gland, and uterus. Only weak signals are present in brain tissue. (C) RT-PCR confirms strong expression of β-netrin RNA in adult mouse kidney, heart, and ovary, as expected from the human blot analyses. In addition, β-netrin RNA is readily detected in several CNS structures including, olfactory bulb and retina. Some expression is observed in other brain regions; however, spinal cord, brain stem, and dorsal root ganglia contain only low levels of β-netrin RNA.

Mentions: RNA expression analysis (Fig. 5A and Fig. B) of β-netrin in human tissues showed that β-netrin is most highly expressed in kidney, spleen, mammary gland, aorta, heart, ovary, prostate, and fetal spleen. Next, we performed semi-quantitative RT-PCR on various mouse tissues and confirmed that, as in humans, β-netrin expression is high in kidneys, hearts, and ovaries (Fig. 5 C). However, we obtained signals from neural tissues as well (Fig. 5 C). A strong signal was obtained from the whole brain (not shown) and retina; fractionation of the whole brain into component regions (Fig. 5 C) demonstrates a low level of expression in most regions, with the exception of the olfactory bulb, where the signal strength approached those obtained from the kidney, heart, and ovary. Below, we have focused our histological studies on those tissues in which PCR signals were high; our studies of the retina will be reported elsewhere.


A novel member of the netrin family, beta-netrin, shares homology with the beta chain of laminin: identification, expression, and functional characterization.

Koch M, Murrell JR, Hunter DD, Olson PF, Jin W, Keene DR, Brunken WJ, Burgeson RE - J. Cell Biol. (2000)

β-Netrin RNA is expressed widely in human and mouse tissues. The same cDNA probe was used to probe adult human tissue blots (A) and a dot blot (B). Only a single major RNA species is detected on the tissue blots. Strong signals are present in spleen, prostate, ovary, heart, kidney and pancreas, mammary gland, and uterus. Only weak signals are present in brain tissue. (C) RT-PCR confirms strong expression of β-netrin RNA in adult mouse kidney, heart, and ovary, as expected from the human blot analyses. In addition, β-netrin RNA is readily detected in several CNS structures including, olfactory bulb and retina. Some expression is observed in other brain regions; however, spinal cord, brain stem, and dorsal root ganglia contain only low levels of β-netrin RNA.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2192657&req=5

Figure 5: β-Netrin RNA is expressed widely in human and mouse tissues. The same cDNA probe was used to probe adult human tissue blots (A) and a dot blot (B). Only a single major RNA species is detected on the tissue blots. Strong signals are present in spleen, prostate, ovary, heart, kidney and pancreas, mammary gland, and uterus. Only weak signals are present in brain tissue. (C) RT-PCR confirms strong expression of β-netrin RNA in adult mouse kidney, heart, and ovary, as expected from the human blot analyses. In addition, β-netrin RNA is readily detected in several CNS structures including, olfactory bulb and retina. Some expression is observed in other brain regions; however, spinal cord, brain stem, and dorsal root ganglia contain only low levels of β-netrin RNA.
Mentions: RNA expression analysis (Fig. 5A and Fig. B) of β-netrin in human tissues showed that β-netrin is most highly expressed in kidney, spleen, mammary gland, aorta, heart, ovary, prostate, and fetal spleen. Next, we performed semi-quantitative RT-PCR on various mouse tissues and confirmed that, as in humans, β-netrin expression is high in kidneys, hearts, and ovaries (Fig. 5 C). However, we obtained signals from neural tissues as well (Fig. 5 C). A strong signal was obtained from the whole brain (not shown) and retina; fractionation of the whole brain into component regions (Fig. 5 C) demonstrates a low level of expression in most regions, with the exception of the olfactory bulb, where the signal strength approached those obtained from the kidney, heart, and ovary. Below, we have focused our histological studies on those tissues in which PCR signals were high; our studies of the retina will be reported elsewhere.

Bottom Line: In addition, beta-netrin is expressed in a limited set of fiber tracts within the brain, including the lateral olfactory tract and the vomeronasal nerve.Functional studies were performed and show that beta-netrin promotes neurite elongation from olfactory bulb explants.Together, these data suggest that beta-netrin is important in neural, kidney, and vascular development.

View Article: PubMed Central - PubMed

Affiliation: Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA.

ABSTRACT
The netrins are a family of laminin-related molecules. Here, we characterize a new member of the family, beta-netrin. beta-Netrin is homologous to the NH(2) terminus of laminin chain short arms; it contains a laminin-like domain VI and 3.5 laminin EGF repeats and a netrin C domain. Unlike other netrins, this new netrin is more related to the laminin beta chains, thus, its name beta-netrin. An initial analysis of the tissue distribution revealed that kidney, heart, ovary, retina, and the olfactory bulb were tissues of high expression. We have expressed the molecule in a eukaryotic cell expression system and made antibodies to the expressed product. Both in situ hybridization and immunohistochemistry were used to describe the cellular source of beta-netrin and where beta-netrin is deposited. beta-Netrin is a basement membrane component; it is present in the basement membranes of the vasculature, kidney, and ovaries. In addition, beta-netrin is expressed in a limited set of fiber tracts within the brain, including the lateral olfactory tract and the vomeronasal nerve. Functional studies were performed and show that beta-netrin promotes neurite elongation from olfactory bulb explants. Together, these data suggest that beta-netrin is important in neural, kidney, and vascular development.

Show MeSH
Related in: MedlinePlus