2Division of Endocrinology, Rutgers- Robert Wood Johnson Medical School, New Brunswick, New Jersey, USA
3Department of Neuroscience and Cell Biology, Rutgers- Robert Wood Johnson Medical School, Piscataway, New Jersey, USA
Methods: Protein-A eluates from plasma of 20 diabetic depression patients and 30 age-matched controls were tested for effects on endothelial cell survival, neurite outgrowth in rat pheochromocytoma (PC12) cells, or process extension and survival in adult rat dentate gyrus neural progenitor cells. The protein-A eluates from depressed or non-depressed, diabetic patients were injected (via intracerebroventricular route) into mice and 7-10 days later behavioral tests (sucrose preference, and tail suspension tests) were conducted to determine whether the autoantibodies induced anhedonia or despair.
Results: Diabetic depression (n=20) autoantibodies caused a significant inhibition of PC12 cell neurite outgrowth (P<0.001) or endothelial cell proliferation compared to autoantibodies in control, diabetic (n=20) or non-diabetic (n=10) patients without depression. Process extension and survival in adult rat dentate gyrus neural progenitor cells was significantly reduced (P<0.001) by diabetic depression autoantibodies (n= 11) compared to the effects from similar concentrations (5-7 μg/mL) of autoantibodies in diabetic (n=12) or non-diabetic patients without depression (n=7). Ten micromolar concentrations of Y27632, a selective Rho-Associated Protein Kinase (ROCK) inhibitor, significantly prevented (P<0.0001) neural progenitor cell process retraction induced by diabetes depression autoantibodies (n=5). Mice treated with diabetic depression autoantibodies (n=16 from two different patients’ autoantibodies) exhibited significantly reduced (P=0.027) sucrose preference (anhedonia) compared to mice treated with diabetic control autoantibodies (n=16 from two different patients’ autoantibodies).
Conclusion: These data suggest that autoantibodies in a subset of older adult diabetic depression inhibit endothelial cell survival, and impair process extension and survival in adult dentate gyrus neural progenitor cells in vitro.
Keywords: Autoantibodies; Diabetes Mellitus; Depression; Neural Progenitor Cells; Neurite Outgrowth
Fibroblast Growth Factor 2 (FGF2) is a broad spectrum angiogenic and neurotrophic growth factor  which does not normally circulate, but is sequestered in extracellular tissue matrices through binding to its low affinity, Heparan Sulfate Proteoglycan (HSPG) co-receptor . Fibroblast growth factor 2 increases in plasma in micro- or albuminuric diabetes  yet low plasma FGF2 levels occurred together with endothelial cell inhibitory autoantibodies in subsets of diabetic macular edema, and /or nephropathy [12,13]. Glomerular loss of HSPG occurs early in diabetic albuminuria  and since HSPG is a known target for autoimmunity , diabetic plasma autoantibodies which target the HSPG, FGF low affinity receptor may disrupt a broad spectrum of FGF2-mediated cellular effects  in neurons and endothelial cells.
The Dentate Gyrus (DG) region in the hippocampus is unique in that it is one of a few brain regions which contain adult neural stem cells . FGF-2 is widely distributed throughout the rat brain , yet it co-localized together with HSPG in only a few regions, including the neurogenic region of the dentate gyrus . Adult neurogenesis in the dentate gyrus has been implicated as an important target for the modulatory effects of stress, exercise, anti-depressant medications or electroconvulsant seizures in human depression or animal models of depressive-like behaviors [19-21]. Major depressive disorder was associated with markedly reduced endogenous FGF expression in mood-regulatory, frontocortical brain regions . In the present study we tested a hypothesis that diabetic plasma IgG autoantibodies increase in subsets of middle-aged or older adult type 2 diabetes suffering with depression, and interfere with the (in vitro) differentiation of adult dentate gyrus neural progenitor cells required for their maturation into adult newborn DG neurons.
We examined plasma autoantibodies in 20 older adults with type 2 diabetic depression, and 30 age-matched (20 diabetic and 10 non-diabetic) non-depressed patients for effects on endothelial cell proliferation, neurite outgrowth in rat pheochromocytoma (PC12) cells, or process extension and survival in adult rat dentate gyrus neural progenitor cells.
Blood drawing: Baseline plasma samples were obtained from study participants prior to the initiation of study treatment in the VADT. Plasma samples obtained at the year 3 study visit were reassessed for endothelial cell autoantibody activity in all five VADT patients having depression. Plasma FGF-2 level was determined with a sensitive, specific two-site IRMA as previously described .
Protein-A affinity chromatography: Protein A chromatography was carried out as previously described . The protein A eluate fractions consisted of total IgG isolated from plasma upon low pH elution from the protein A column. The ‘active protein A eluate’ is defined as the protein A eluate which caused significant inhibitory activity in endothelial cell survival assays.
Endothelial cell survival assay: Bovine pulmonary artery endothelial cells (Clonetics, Inc. San Diego, CA) were grown in Medium 199 plus 10% fetal calf serum and endothelial cell growth medium (EGM, Clonetics, Inc., San Diego, CA). Endothelial cell number assays were carried out as previously reported . Confluent cells were trypsinized and plated at 1 to 10 x 103 cells/ well in Medium 199 plus 10% fetal calf serum in 96-well plates. After 1 or 2 days incubation for cells to reach 60 to 80% confluency, test fractions (30 μg/mL of protein A eluates of plasma) were added to wells in quadruplicate. After 48 hours’ incubation in the presence of test fractions, cells were washed with PBS and processed for the colorimetric estimation of cell number, i.e. cellassociated acid phosphatase activity, as previously described . There was a linear relationship between endothelial cell number and optical density at 410 nm as previously described . Growth-promoting activity is expressed as a percentage of the control cell number for cells grown in the absence of test protein-A eluate fractions.
Neurite outgrowth inhibition assays: Undifferentiated rat pheochromocytoma PC12 cells obtained from ATCC (Manassa, VA) were grown in DMEM containing 10% horse serum, 5% fetal calf serum and 10 ng/mL bFGF was added in order to induce neuronal differentiation. Test protein-A eluate fractions (1:50 dilution = 30 μg/mL protein) were added to cells in duplicate or triplicate and incubated at 37°C for 48 hrs. Next the proportion of cells expressing a neurite of > 2 cell diameters in length was counted and compared to the proportion of bFGF- stimulated neurite cell expression in dishes containing bFGF without added test fractions as previously described .
Adult neural progenitor cells: The stem cells were originally isolated from the dentate gyrus of adult Fisher 344 rats and were obtained from Dr. Fred Gage at the Salk Institute. The expanded cultures from single clones were infected with retrovirus to express GFP and selected as previously described . The NPCs were plated at 15,000 cells/cm2 on poly-L-ornithine and laminin in Dulbecco’s Modified Eagle Medium (DMEM)/F12 medium high glucose (Omega Scientific, Tarzana, CA) containing N2 supplement (In vitrogen, Grand Island, NY), L-glutamine (2mM), penicillin/ streptomycin (100 U/ml) (Medium A) and FGF-2 (20ng/ml, PeproTech, Rocky Hill, NJ). Upon reaching 70-90% confluency, NPCs were trypsinized with 0.05% trypsin EDTA and plated at 144,500 cells/35mm dish in medium A without FGF-2. Cultures of GFP+ stem cells between passages 15 and 20 were used in this study. Test protein-A eluate fractions (5-7 μg/mL protein) were added to cells on DIV 3 in duplicate or triplicate and incubated at 37°C for 4-6 days. On DIV 7, the proportion of cells expressing a neurite of > 2 cell diameters in length was counted and compared to the proportion of cells expressing processes in dishes without added test fractions. NPC cell survival was assessed on DIV 9 as the proportion in two groups of 50–150 cells/dish that excluded 0.04% trypan blue.
Sucrose preference test: One week following antibody infusions, anhedonia was measured using the sucrose preference test in each mouse’s home cage. Two different autoantibodies in each subgroup of diabetic depression or diabetic control patients were tested on a total of 16 mice. Prior to testing, each mouse was acclimated to a 1% sucrose solution for 24 hours, via two- 50 mL tubes with sipper tops. Following the completion of the sucrose preference pretrial, two days of sucrose preference were measured. During this time, each mouse had access to a sipper tube containing water and another containing 1 % sucrose water. After one day, the locations of the bottles were switched to account for potential side preferences. All bottles are massed before and after each day of testing. Sucrose preference is equal to the ratio of total sucrose solution consumed to overall total solution consumed.
Tail suspension test: One day following the completion of sucrose preference testing (ten days following antibody infusions) behavioral despair was measured using the tail suspension test. One autoantibody in each subgroup of diabetic depression or diabetic control patients was tested on a total of 10 mice. The open-end of a 25 mL pipette was firmly secured to a metal shelf raised one-half meters above the bench top using packing tape. A water bath was positioned below the area of testing as a safety precaution. The final centimeter of each mouse-tail was secured to the closed-end of the pipette using ≈ 2 centimeters of masking tape. Trial duration was six-minutes long. All trials were videotaped and immobile behavior during the final four minutes was measured manually by viewing the recording at a later time.
Depression No Depression P-Value*
DM dur (yrs)
Insulin use (%)
60.7+ 11.0 68.4 + 10.9 0.154
9.2 + 1.4 8.6 + 1.6 0.174
19.4 + 11.7 13.4 + 11.1 0.117
33.1 + 6.1 31.1 + 5.3 0.306
85 50 0.016^
29.0 + 4.3**
Depression No Depression P-Value*
ME, AMD (%)
Painful neur (%)
Inhib EC act (%)
Low FGF-2 (%) ^
80 40 0.0098
50 15 0.018
70 40 0.057
75 65 0.490
88 65 0.101
Diabetic depression autoantibodies (5-7 μg/mL protein) substantially inhibited NPC branching process extension, and also caused substantial contraction in NPC cells [arrows, Figure 2f] compared to the morphology in NPC cells cultured without autoantibodies [Figure 2d or incubated with similar concentrations of protein A eluates from diabetes without depression [Figure 2e].
We next tested for involvement of sulfated proteoglycans or heparin GAG in the maintenance of a differentiated, neuriticallyextended phenotype in NPC cells cultured without FGF-2. Sodium chlorate (5 mM) caused NPC dendrites to rapidly detach from the matrix and degenerate. At higher sodium chlorate concentrations (10-30 mM), NPC cell bodies also rapidly contracted, detached from the matrix and died. Ten microgram/mL concentrations of a specific (6-0,2-0) densely sulfated, heparin disaccharide caused NPC neurites to retract, and shrink by up to 50% after five minutes. After an additional five minutes’ exposure, highly branched NPC cells neurites had acquired a ‘dystrophic’ morphology without effects on NPC cell body morphology. On the other hand, five-ten minutes’ exposure to (6-0,2-0) de-sulfated heparin disaccharide (10 μg/mL) caused NPC neurite- lengthening without any discernable effect on NPC cell body morphology (data not shown). The simultaneous addition of 10 μg/mL concentrations of (6-0,2- 0) densely sulfated, heparin disaccharide together with (5 μg/mL) diabetic depression autoantibodies caused neurite retraction, dendritic simplification and distal neurite degeneration (after 10 minutes exposure), the latter was characterized by vacuolization, beading and distal neuritic detachment from the matrix. Diabetic depression autoantibodies (5 μg/mL) alone caused NPC cell contraction and modest (15-25%) neurite retraction within 10 minutes of addition to the cells.
The association reported here between highly potent anti-EC autoantibodies and co-morbid depression is striking and novel. Adult dentate gyrus neural progenitor cells develop in a vascular microenvironment enriched in angiogenic growth factors  suggesting they may be a target for circulating anti-endothelial, anti-neuronal autoantibodies. Our finding that maculopathy and nephropathy were increased in diabetic depression agrees with previous reports . Since potent diabetic anti-EC autoantibodies were previously demonstrated to activate Rho A/Rho kinase signaling in EC , and Rho A/Rho kinase activation is associated with endothelial cell barrier dysfunction , diabetic plasma autoantibodies which can target endothelial or endothelial-like, vascular cells (i.e. adult neural stem cells)  may not only have a causative role in mediating increased capillary permeability underlying diabetic macular edema and/or nephropathy, but also in subsets of co-morbid depression occurring together with these microvascular complications.
Prior findings and the current data strongly suggest that subsets of diabetic depression autoantibodies target cell surface HSPG abundant on neurons and endothelial cells. First, the diabetic maculopathy/nephropathy autoantibodies bound with specific, high affinity to heparin Sepharose columns . Second, they displayed increased binding to PC-12 cell-derived purified HSPG . Third, the proteoglycan sulfation inhibitor, chlorate, abrogated autoantibody-induced PC12 neurite retraction . Fourth, the autoantibodies caused endothelial cell contraction, loss of cell attachment and apoptosis by a mechanism involving activation of RhoA/Rho kinase, increased intracellular Ca2+ release, and caspase activation . Taken together with the present finding that diabetic depression autoantibodies caused contraction and decreased NPC survival (effects which were mimicked by chlorate ion), diabetic depression autoantibodies may target sulfated proteoglycans on the NPC cell surface involved in mediating attachment to heparin-binding substances in the underlying matrix.
NPC neuritic retraction vs outgrowth was differentially modulated by densely- sulfated or unsulfated forms of a heparin disaccharide, respectively. These findings may be consistent with the reported effects (on neurite outgrowth) of substances which inhibit or enhance the interaction between (NPC) cell surface HSPG and matrix-associated, heparin sulfate- or heparan sulfate- binding substances . Antibodies which cross-linked cell surface HSPG were reported to induce Rhodependent cytoskeleton changes, e.g. in actin stress fibers, mediating fibroblast cell shape changes . Since Rho A/Rho kinase signaling activation in neurons also mediates dendritic simplification and neuritic retraction , our finding that a selective Rho kinase inhibitor interfered with neurite retraction and NPC cell killing by diabetic depression autoantibodies suggests possible involvement of ROCK activation in the mechanism of autoantibody-mediated effects in NPCs.
The precise identity and tissue origin(s) of the auto-antigens targeted by diabetic depression autoantibodies is unknown. TNF-α induced endothelial cells to express a specific (N-sulfated, 2-O, 6-O sulfated)3 heparan sulfate domain important for mediating FGF-2 and FGFR binding . Of interest, phage display-derived antibodies specific for the same, densely sulfated heparan GAG domain induced apoptosis in lung epithelial cells  consistent with the possibility that tissue fate specification is mediated in part by HSPG heparan sulfate microstructural variation. The fine specificity in HSPG GAG saccharide structures is thought to play an important role in neuronal development by modulating interactions between cell surface receptors and heparin binding growth factors whose spatio-temporal expression is tightly developmentally-regulated . More study is needed to determine whether autoantibodies arising under pro-inflammatory conditions such as poorly-controlled diabetes which target HSPGs elaborated from peripheral tissues, e.g. glomerular capillary, cross-react with HSPGs expressed in certain neuronal cells.
Even though adult, obese type 2 diabetes is not a systemic autoimmune disease, the prevalence of anti-EC autoantibodies (30%) in a VADT, obese type 2 DM cohort comprised of 182 older adults  was similar to that which was previously reported in adult type 1 diabetes . More potent anti-EC autoantibodies were found in a higher percentage (38%) or 117/305 proteinuric, adult type 2 diabetic nephropathy from the VA-NEPHRON D trial . Baseline insulin or fibrate medication use were risk factors significantly inversely associated with the baseline presence of anti-EC autoantibodies in the VADT cohort  consistent with a possible role for up- or down-regulation of EC heparanase secretion (in vitro) by pro-inflammatory cytokines  or by insulin , respectively in the elaboration of EC-derived HSPG.
Adult rat or mouse NPCs both express HSPG required for FGF- 2 mediated proliferation  with substantially higher HPSG expression occurring in adult rat NPCs . Whether decreased neurogenesis plays a role in the induction of depressive-like behavior, or might be preventable through administration of a selective ROCK inhibitor are unknown and require further study. The current data might be consistent with prior reports indicating that Rho A activation in adult hippocampal neural progenitor cells suppressed neurogenesis . It is not known, however, whether suppression of neurogenesis by ROCK-activating autoantibodies might account in part for the observation that many of the diabetic depression patients in the current study cohort were poorlyresponsive to treatment with SSRI anti-depressant medications.
A limitation of our study is that the results may only apply to older men with longstanding type 2 diabetes who had a substantial burden of microvascular diabetic complications. More study is needed in other populations including women with diabetes. Anti-endothelial, anti-neuronal autoantibodies may result from tissue injury and have only a limited bystander role in depression. Yet our preliminary results indicate persistence of depressive symptoms in 4/4 diabetic patients in whom plasma autoantibody persisted for three years or longer. On the other hand, spontaneous disappearance of autoantibodies was associated with excellent treatment response to a selective serotonin reuptake inhibitor medication in a 72 year- old man with major depression.
Depression occurring in older adults remains underrecognized and undertreated . A much larger randomized study would be required to test whether NPC-targeting autoantibodies might be useful as a biomarker for the early detection of depression subsets. Despite accumulating evidence linking alterations in the rate of adult dentate gyrus hippocampal neurogenesis with depression, the function of newborn adult DG neurons in hippocampus networks remains poorly understood . Since immature adult dentate gyrus neurons were reported to be hyper-excitable compared to mature DG neurons  one possibility is that DG NPCs might have increased susceptibility to autoantibody-induced excitotoxicity, as was reported previously in rat hippocampal neurons exposed to low concentrations of cancer fatigue/depression autoantibodies .
The authors report no conflict of interest that would affect the objectivity of the findings presented.
- Banasr M, Dwyer JM, Duman RS. Cell atrophy and loss in depression: reversal by antidepressant treatment. Curr Opin Cell Biol. 2011; 23(6):730-737. doi: 10.1016/j.ceb.2011.09.002.
- Aziz R, Steffens DC. What are the causes of late-life depression? Psychiatr Clin North Am. 2013; 36(4):497-516. doi: 10.1016/j. psc.2013.08.001.
- Sullivan MD, O’Connor P, Feeney P, Hire D, Simmons DL, Raisch DW, et al. Depression predicts all-cause mortality: epidemiological evaluation from the ACCORD HRQL substudy. Diabetes Care. 2012; 35(8):1708- 15. doi: 10.2337/dc11-1791.
- Lin EH, Rutter CM, Katon W, Heckbert SR, Ciechanowski P, Oliver MM, et al. Depression and advanced complications of diabetes: a prospective cohort study. Diabetes Care. 2010; 33(2):264-9. doi: 10.2337/dc09-1068.
- Holt RIG, de Groot M, Lucki I, Hunter CM, Sartorius N, Golden SH. NIDDK International Conference Report on Diabetes and Depression: Current Understanding and Future Directions. Diabetes Care. 2014; 37(8):2067-2077. doi: 10.2337/dc13-2134.
- Anderson RJ, Freedland KE, Clouse RE, Lustman PJ The prevalence of comorbid depression in adults with diabetes: a meta-analysis. Diabetes Care 2001; 24(6):1069–1078.
- Yu MK, Katon W, Young BA. Diabetes self-care, major depression, and chronic kidney disease in an outpatient diabetic population. Nephron Clin Pract. 2013; 124(1-2):106-112. doi: 10.1159/000355551.
- Deckert T, Feldt-Rasmussen B, Borch-Johnsen K, Jensen T, Kofoed- Enevoldsen A. Albuminuria reflects widespread vascular damage. The Steno hypothesis. Diabetologia. 1989; 32(4):219-226.
- Gospodarowicz D, Ferrara N, Schweigerer L, Neufeld G. Structural characterization and biological functions of fibroblast growth factor. Endocr Rev. 1987; 8(2):95-114.
- Vlodavsky I, Fuks Z, Ishai-Michaeli R, Bashkin P, Levi E, Korner G, et al. Extracellular matrix-resident basic fibroblast growth factor: implication for the control of angiogenesis. J Cell Biochem. 1991; 45(2):167-176.
- Zimering MB, Eng J. Increased basic fibroblast growth factor-like substance in plasma from a subset of middle-aged or elderly male diabetic patients with microalbuminuria or proteinuria. J Clin Endocrinol Metab. 1996; 81(12):4446-4452.
- Zimering MB, Anderson RJ, Moritz TE, Ge L; Investigators for the VADT. Endothelial cell inhibitory autoantibodies are associated with laser photocoagulation in adults from the Veterans Affairs Diabetes Trial. Metabolism. 2009; 58(6):882-887. doi: 10.1016/j. metabol.2009.02.023.
- Zimering MB, Alder J, Pan Z, Donnelly RJ. Anti-endothelial and antineuronal effects from auto-antibodies in subsets of adult diabetes having a cluster of microvascular complications. Diabetes Res Clin Pract. 2011; 93(1):95-105. doi: 10.1016/j.diabres.2011.03.029.
- Wijnhoven TJ, van den Hoven MJ, Ding H, van Kuppevelt TH, van der Vlag J, Berden JH, et al. Heparanase induces a differential loss of heparan sulphate domains in overt diabetic nephropathy. Diabetologia. 2008; 51(2):372-382.
- Renaudineau Y, Révélen R, Dueymes M, Levy Y, Youinou P. Autoantibodies to heparan sulfate proteoglycans. Autoimmun Rev. 2002; 1(5):305-312.
- Palmer TD, Takahashi J, Gage FH. The adult rat hippocampus contains primordial neural stem cells. Mol Cell Neurosci. 1997; 8(6):389-404.
- Fuxe K, Tinner B, Zoli M, Pettersson RF, Baird A, Biagini G, et al. Computer-assisted mapping of basic fibroblast growth factor immunoreactive nerve cell populations in the rat brain. J Chem Neuroanat. 1996; 11(1):13-35.
- Fuxe K, Chadi G, Tinner B, Agnati LF, Pettersson R, David G. On the regional distribution of heparan sulfate proteoglycan immunoreactivity in the rat brain. Brain Res. 1994; 636(1):131-138.
- Warner-Schmidt JL, Duman RS. Hippocampal neurogenesis: opposing effects of stress and antidepressant treatment. Hippocampus. 2006; 16(3):239-249.
- Newton SS, Collier EF, Hunsberger J, Adams D, Terwilliger R, Selvanayagam E, et al. Gene profile of electroconvulsive seizures: induction of neurotrophic and angiogenic factors. J Neurosci. 2003; 23(34):10841-10851.
- Fuchs E. Social stress in tree shrews as an animal model of depression: an example of a behavioral model of a CNS disorder. CNS Spectr. 2005; 10(3):182-190.s
- Evans SJ, Choudary PV, Neal CR, Li JZ, Vawter MP, Tomita H, et al. Dysregulation of the fibroblast growth factor system in major depression. Proc Natl Acad Sci USA. 2004; 101(43):15506–15511.
- Brody BL, Gamst AC, Williams RA, Smith AR, Lau PW, Dolnak D, et al. Depression, visual acuity, comorbidity, and disability associated with age-related macular degeneration. Ophthalmology. 2001; 108(10):1893-1900.
- Zimering MB, Anderson RJ, Moritz TE, Ge L; Investigators for the VADT. Low plasma basic fibroblast growth factor is associated with laser photocoagulation treatment in adult type 2 diabetes mellitus from the Veterans Affairs Diabetes Trial. Metabolism. 2009; 58(3):393-400. doi: 10.1016/j.metabol.2008.10.014.
- Zimering MB, Thakker-Varia S. Increased fibroblast growth factorlike autoantibodies in serum from a subset of patients with cancerassociated hypercalcemia. Life Sci. 2002; 71(25):2939-2959.
- Song HJ, Stevens CF, Gage FH. Neural stem cells from adult hippocampus develop essential properties of functional CNS neurons. Nat Neurosci. 2002; 5(5):438–445.
- Bartolomucci A, Possenti R, Levi A, Pavone F, Moles A. The role of the vgf gene and VGF-derived peptides in nutrition and metabolism. Genes Nutr. 2007; 2(2):169-180. doi: 10.1007/s12263-007-0047-0.
- Palmer TD, Ray J, Gage FH. FGF-2-responsive neuronal progenitors reside in proliferative and quiescent regions of the adult rodent brain. Mol Cell Neurosci. 1995; 6(5):474-486.
- Keller KM, Brauer PR, Keller JM. Modulation of cell surface heparan sulfate structure by growth of cells in the presence of chlorate. Biochemistry. 1989; 28(20):8100-8107.
- Zimering, M. B., and Pan, Z. Autoantibodies in type 2 diabetes induce stress fiber formation and apoptosis in endothelial cells. J Clin Endocrinol Metab. 2009; 94(6):2171-2177. doi: 10.1210/jc.2008- 2354.
- Palmer TD, Willhoite AR, Gage FH. Vascular niche for adult hippocampal neurogenesis. J Comp Neurol. 2000; 425(4):479-494.
- van Nieuw Amerongen GP, Beckers CM, Achekar ID, Zeeman S, Musters RJ, van Hinsbergh VW. Involvement of Rho kinase in endothelial barrier maintenance. Arterioscler Thromb Vasc Biol 2007; 27(11):2332–2339.
- Zimering MB, Moritz TE, Donnelly RJ. Anti-neurotrophic effects from autoantibodies in adult diabetes having primary open angle glaucoma or dementia. Front Endocrinol (Lausanne). 2013; 4:58. doi: 10.3389/ fendo.2013.00058.
- Kinnunen A, Niemi M, Kinnunen T, Kaksonen M, Nolo R, Rauvala H. Heparan sulphate and HB-GAM (heparin-binding growth-associated molecule) in the development of the thalamocortical pathway of rat brain. Eur J Neurosci. 1999; 11(2):491-502.
- Saoncella S, Echtermeyer F, Denhez F, Nowlen JK, Mosher DF, Robinson SD, et al. Syndecan-4 signals cooperatively with integrins in a Rho-dependent manner in the assembly of focal adhesions and actin stress fibers. Proc Natl Acad Sci USA. 1999; 96(6):2805-2810.
- Katoh H, Aoki J, Ichikawa A, Negishi M. p160 RhoA-binding kinase ROKalpha induces neurite retraction. J Biol Chem. 1998; 273(5):2489- 2492.
- Smits NC, Kurup S, Rops AL, ten Dam GB, Massuger LF, Hafmans T, et al. The heparan sulfate motif (GlcNS6S-IdoA2S)3, common in heparin, has a strict topography and is involved in cell behavior and disease. J Biol Chem. 2010; 285(52):41143-41151. doi: 10.1074/jbc. M110.153791.
- Wangel AG, Kontiainen S, Scheinin T, Schlenzka A, Wangel D, Mäenpää J. Anti-endothelial cell antibodies in insulin-dependent diabetes mellitus. Clin Exp Immunol. 1992; 88(3):410-413.
- Zimering MB, Zhang JH, Guarino PD, Emanuele N, McCullough PA, Fried LF; Investigators for the VA NEPHRON-D. Endothelial cell autoantibodies in predicting declining renal function, end-stage renal disease, or death in adult type 2 diabetic nephropathy. Front Endocrinol (Lausanne). 2014; 5:128. doi: 10.3389/fendo.2014.00128.
- Chen G, Wang D, Vikramadithyan R, Yagyu H, Saxena U, Pillarisetti S, et al. Inflammatory cytokines and fatty acids regulate endothelial cell heparanase expression. Biochemistry. 2004; 43(17):4971-4977.
- Han J, Woytowich AE, Mandal AK, Hiebert LM. Heparanase upregulation in high glucose-treated endothelial cells is prevented by insulin and heparin. Exp Biol Med (Maywood). 2007; 232(7):927-934.
- Ray J, Gage FH. Differential properties of adult rat and mouse brainderived neural stem/progenitor cells. Mol Cell Neurosci. 2006; 31(3):560-573.
- Mellough CB, Cho S, Wood A, Przyborski S. Neurite formation by neurons derived from adult rat hippocampal progenitor cells is susceptible to myelin inhibition. Neurochem Int. 2011; 59(3):333- 340. doi: 10.1016/j.neuint.2011.01.015.
- Webber AP, Martin JL, Harker JO, Josephson KR, Rubenstein LZ, Alessi CA. Depression in older patients admitted for postacute nursing home rehabilitation. J Am Geriatr Soc. 2005; 53(6):1017-1022.
- Piatti VC, Ewell LA, Jill K. Leutgeb JK. Neurogenesis in the dentate gyrus: carrying the message or dictating the tone. Front Neurosci. 2013; 7:50. doi: 10.3389/fnins.2013.00050.