Expression of the 67 kDa laminin receptor (67LR) during retinal development: correlations with angiogenesis.
Interaction of vascular cells with the laminin component of basement membranes is important for normal cell function. Likewise, abnormal interactions may have a critical role in vascular pathology. It has been previously demonstrated that the 67 kDa laminin receptor (67LR) is expressed at high levels during proliferative retinopathy in a mouse model and in the current study we have examined 67LR in the neonatal mouse to determine if this receptor plays a role in aspects of developmental angiogenesis in the developing murine retina. Groups of C57/BL6 mice were killed at postnatal day P1, P3, P5, P7, P9 and P11 to assess the retinal vasculature. A number of mice were perfused with FITC-dextran and the eyes removed, fixed in 4% paraformaldehyde (PFA) and flat-mounted for confocal scanning laser microscopy. The eyes from the remaining mice were either placed in 4% PFA and embedded in paraffin-wax, or had the neural retina dissected off and total RNA or protein extracted. Immunofluorescence, in situ hybridization, quantitative reverse transcriptase polymerase chain reaction and Western blotting analysis were employed to locate and determine expression levels of 67LR. Both 67LR mRNA and protein expression showed a characteristic bi-phasic expression pattern which correlated with key stages of retinal vascular development in the murine retina. 67LR showed high expression levels at P1 (P < 0.05) (correlating with superficial vascular plexus formation) and at P7 (P < 0.05) (correlating with deep vascular plexus formation). Conversely, 67LR expression was decreased when active angiogenic activity was lowest. Significantly, optical sectioning of retinal flat-mounts revealed high levels of 67LR expression in developing segments of both superficial and deep capillary plexi, a pattern which co-localized strongly with laminin. 67LR is regulated during post-natal development of the retinal vasculature. High levels of 67LR during the two well-defined phases of retinal capillary plexus formation suggests that this receptor may play an important role in retinal angiogenesis.
McKenna
D J
DJ
Department of Ophthalmology, The Queen's University of Belfast, The Royal Victoria Hospital, Belfast BT12 6BA, Northern Ireland, U.K.
Simpson
D A
DA
Feeney
S
S
Gardiner
T A
TA
Boyle
C
C
Nelson
J
J
Stitt
A W
AW
D000818:Animals; D000831:Animals, Newborn; D015139:Blotting, Southern; D015153:Blotting, Western; D003911:Dextrans / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005451:Fluorescein Angiography; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D017403:In Situ Hybridization; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D018919:Neovascularization, Physiologic; D016612:Paraffin Embedding; D012333:RNA, Messenger; D017474:Receptors, Laminin / Q000378:metabolism*; D012160:Retina / Q000254:growth & development* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003911:Dextrans; D012333:RNA, Messenger; D017474:Receptors, Laminin; D016650:Fluorescein-5-isothiocyanate