Delineating multiple functions of VEGF-A in the adult brain

Vascular endothelial growth factor-A (abbreviated throughout this review as VEGF) is mostly known for its angiogenic activity, for its activity as a vascular permeability factor, and for its vascular survival activity [1]. There is a growing body of evidence, however, that VEGF fulfills additional less ‘traditional’ functions in multiple organs, both during development, as well as homeostatic functions in fully developed organs. This review focuses on the multiple roles of VEGF in the adult brain and is less concerned with the roles played by VEGF during brain development, functions described elsewhere in this review series. Most functions of VEGF that are essential for proper brain development are, in fact, dispensable in the adult brain as was clearly demonstrated using a conditional brain-specific VEGF loss-of-function (LOF) approach. Thus, in contrast to VEGF LOF in the developing brain, a process which is detrimental for the growth and survival of blood vessels and leads to massive neuronal apoptosis [2–4], continued signaling by VEGF in the mature brain is no longer required for maintaining already established cerebral vasculature and its inhibition does not cause appreciable vessel regression, hypoxia or apoptosis [4–7]. Yet, VEGF continues to be expressed in the adult brain in a constitutive manner. Moreover, VEGF is expressed in the adult brain in a region-specific manner and in distinctive spatial patterns incompatible with an angiogenic role (see below), strongly suggesting angiogenesis-independent and possibly also perfusion-independent functions. Here we review current knowledge on some of these ‘non-traditional’, often unexpected homeostatic VEGF functions, including those unrelated to its effects on the brain vasculature. These effects could be mediated directly (on non-vascular cells expressing cognate VEGF receptors) or indirectly (via the endothelium). Experimental approaches aimed at distinguishing between these possibilities for each particular VEGF function will be described. This review is only concerned with homeostatic functions of VEGF in the normal, non-injured brain. The reader is referred elsewhere in this series for a review on VEGF actions in response to various forms of brain injury and/or brain pathology.     

Aneuploidy in the normal and diseased brain

The brain is remarkable for its complex organization and functions, which have been historically assumed to arise from cells with identical genomes. However, recent studies have shown that the brain is in fact a complex genetic mosaic of aneuploid and euploid cells. The precise function of neural aneuploidy and mosaicism are currently being examined on multiple fronts that include contributions to cellular diversity, cellular signaling and diseases of the central nervous system (CNS). Constitutive aneuploidy in genetic diseases has proven roles in brain dysfunction, as observed in Down syndrome (trisomy 21) and mosaic variegated aneuploidy. The existence of aneuploid cells within normal individuals raises the possibility that these cells might have distinct functions in the normal and diseased brain, the latter contributing to sporadic CNS disorders including cancer. Here we review what is known about neural aneuploidy, and offer speculations on its role in diseases of the brain. Received 13 April 2006; received after revision 2 June 2006; accepted 13 July 2006     

Morphology of the carnation-light synthetic lethal focus in Drosophila melanogaster

A histological study of the carnation-light lethal focus revealed morphological abnormalities in brain tissue. The ratio of core width to total brain width and brain texture consistently differed between lethal (car-lt) and their non-lethal sibs.     

Effect of diabetes on the enzymes of the cholinergic system of the rat brain

Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activities were determined in several brain regions of normal and streptozotocin-induced diabetic rats. The diabetic rats exhibited significant increase in ChAT activity (p less than 0.05) in all brain regions studied except for the cortex and the midbrain. Meanwhile, the diabetes condition was associated with significant increase (p less than 0.05) in AChE activity of the bulbus olfactorius, medulla oblongata and cerebellum. These data suggest that uncontrolled diabetes is associated with significant alterations in the brain cholinergic systems.     

The effect of proctolin on the adenylate and guanylate cyclases in the Locusta brain at various developmental stages.

Proctolin at concentrations 10(-8)-10(-7) M elevated by 40% brain adenylate cyclase activity of adult Locusta migratoria migratoriodes R.F. In moulting individuals, proctolin caused a decrease in brain adenylate cyclase activity, and it proved to be ineffective in the larvae. Proctolin caused only a slight decrease on guanylate cyclase activity of the brain at every developmental stage.     

Cholinergic, monoaminergic and glutamatergic changes following perinatal asphyxia in the rat

Perinatal asphyxia (PA) is considered to lead to a variety of brain disorders including spasticity, epilepsy, mental retardation, and minimal brain disorder syndromes and may form the basis for psychiatric and neurodegenerative diseases later in life. We examined markers for neuronal transmission involved in the pathomechanisms of PA and candidates as mediators for long-term sequelae. We tested tyrosine hydroxylase (TH) and the vesicular monoamine transporter (VMAT) representing the monoaminergic system, the vesicular acetylcholine transporter (VAChT), and the excitatory amino acid carrier 1 (EAAC1), a neuronal subtype of the glutamate transporter, using immunohistochemistry on brain sections of rats subjected to graded PA. Three months following the asphyxiant insult immunoreactive (IR)-TH was decreased in striatum, hippocampus, thalamus, frontal cortex, and cerebellum; IR-VMAT was increased, and IR-VAChT was decreased in striatum. IR-EAAC1 glutamate transporter was increased in frontal cortex. The cholinergic, monoaminergic, and glutamatergic changes, still observed 3 months after the asphyxiant insult, may reflect their involvement in the pathomechanisms of PA and indicate mechanisms leading to long-term complications of PA. The variable consequences on the individual markers in several brain regions may be explained by specific susceptibility of cholinergic, monoaminergic, and glutamatergic neurons to the asphyxiant insult. Received 16 March 1999; received after revision 20 May 1999; accepted 8 July 1999     

Changes in the G6PDH/6PGDH ratio in the chick brain during development

Summary The profile of the G6PDH/6PGDH ratio at various stages of development was drawn on the basis of the specific activities of glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconic dehydrogenase (6PGDH) found in the embryonic brain of the chick. The ratio value lower than 1, found in the adult chick brain, is a special biochemical feature which emerges at a certain time-point of development, occurring around the middle of the prehatching age.Acknowledgment. This work was supported by grants from M.P.I. and C.N.R.     

Immunoreactive luteinizing hormone-containing neurons in the brain of the white-footed mouse,Peromyscus leucopus

Summary The distribution of immunoreactive LH in the brain of the white-footed mouse (Peromyscus leucopus) was determined using immunocytochemical procedures. Immunoreactive fibers are located in the hypothalamus, preoptic area, septum and amygdala. Stained cell bodies are seen in the arcuate nucleus and preoptic area. Gonadectomy enhances staining for LH in the brain.We wish to thank the National Institutes of Arthritis, Metabolism, and Digestive Diseases (NIAMDD) for the gift of the antiserum to LH.     

Effect of diabetes on the enzymes of the cholinergic system of the rat brain

Summary Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activities were determined in several brain regions of normal and streptozotocin-induced diabetic rats. The diabetic rats exhibited significant increase in ChAT activity (p<0.05) in all brain regions studied except for the cortex and the midbrain. Meanwhile, the diabetes condition was associated with significant increase (p<0.05) in AChE activity of the bulbus olfactorius, medulla oblongata and cerebellum. These data suggest that uncontrolled diabetes is associated with significant alterations in the brain cholinergic systems.To whom requests of reprints should be addressed.This work was supported by grants from the National Aeronautics and Space Administration (NSG 2183 and NAG-2-411), a grant from the National Institutes of Health (NIH Grant RR0811) and a grant from the Division of Research Resources, National Institutes of Health (NIH Grant RR03020).     

Emerging concepts in acute mountain sickness and high-altitude cerebral edema: from the molecular to the morphological

Acute mountain sickness (AMS) is a neurological disorder that typically affects mountaineers who ascend to high altitude. The symptoms have traditionally been ascribed to intracranial hypertension caused by extracellular vasogenic edematous brain swelling subsequent to mechanical disruption of the blood–brain barrier in hypoxia. However, recent diffusion-weighted magnetic resonance imaging studies have identified mild astrocytic swelling caused by a net redistribution of fluid from the “hypoxia-primed” extracellular space to the intracellular space without any evidence for further barrier disruption or additional increment in brain edema, swelling or pressure. These findings and the observation of minor vasogenic edema present in individuals with and without AMS suggest that the symptoms are not explained by cerebral edema. This has led to a re-evaluation of the relevant pathogenic events with a specific focus on free radicals and their interaction with the trigeminovascular system. (Part of a multi-author review.)     

Fixation of polyunsaturated fatty acids by alpha fetoprotein and serum albumin in rats. Comparison with the accumulation of these acids in developing rat brain]

Quantitative data are presented on the fatty acid composition of rat alpha-fetoprotein (AFP) and serum albumin, (SA), and of brain extracts of suckling rats. In AFP and SA preparations, 40% and 13%, respectively, of total fatty acids present are polyenoic acids. Among them, docosahexaenoic acid is quantitatively the most important in AFP, while in SA, arachidonic acid is largely predominant. Both docosahexaenoic and arachidonic acids were the predominant polyenoic acids in brain extracts. The rate of accumulation of these acids in the brain of suckling rats and the rate of AFP secretion during the same period showed a maximum around 10--12 days after birth. These results suggest that AFP and SA play an important role in the transport and the incorporation of polyunsaturated fatty acids in the developing brain.     

Evaluation of monoaminergic receptors in the genetically epilepsy prone rat

The intensity of sound-induced convulsions in the genetically epilepsy-prone rat (GEPR) was reduced in a dose related fashion by intracerebroventricular administration of dobutamine, (beta 1 agonist), terbutaline (beta 2 agonist) or phenylephrine (alpha 1 agonist). BHT-920 (alpha 2 agonist) did not cause a dose-related decrease in sound-induced convulsion intensity. Binding studies showed that whole brain alpha and beta receptor densities (Bmax) were normal while the Kd was increased for the beta ligand in GEPR brain.     

Increased contents of phenylethanolamine, m-octopamine and p-octopamine in the hypothalamus and brain stem of spontanously hypertensive rats (S.H.R. Kyoto)]

Phenylethanolamine, p-octopamine and m-octopamine contents were determined in the hypothalamus and the brain stem of spontaneously hypertensive Rats (S.H.R. Kyto) and the corresponding controls (Wistar Kyoto). In three-week-old Rats, phenylethanolamine and p-octopamine are found to be present in S.H.R. hypothalamus and brain stem at concentrations twice as high as compared to Wistar Kyoto Rats. The amount of m-octopamine is 5-fold higher in the brain stem of S.H.R. Rats as compared to Wistar Kyoto Rats.     

Rate of tryptophan hydroxylation in vivo in brain nuclei of genetically hypertensive rats of the Lyon strain

The rate of tryptophan hydroxylation in vivo is unaltered in brain areas of 5, 9 and 21 week-old Lyon genetically Hypertensive (LH) rats as compared to both Lyon Normotensive (LN) and Low Blood Pressure (LL) rats, except for a decrease in the C1 area of the medulla oblongata in 9 week-old animals.     

Developmental and pathological angiogenesis in the central nervous system

Angiogenesis, the formation of new blood vessels from pre-existing vessels, in the central nervous system (CNS) is seen both as a normal physiological response as well as a pathological step in disease progression. Formation of the blood–brain barrier (BBB) is an essential step in physiological CNS angiogenesis. The BBB is regulated by a neurovascular unit (NVU) consisting of endothelial and perivascular cells as well as vascular astrocytes. The NVU plays a critical role in preventing entry of neurotoxic substances and regulation of blood flow in the CNS. In recent years, research on numerous acquired and hereditary disorders of the CNS has increasingly emphasized the role of angiogenesis in disease pathophysiology. Here, we discuss molecular mechanisms of CNS angiogenesis during embryogenesis as well as various pathological states including brain tumor formation, ischemic stroke, arteriovenous malformations, and neurodegenerative diseases.     

Immunoreactive luteinizing hormone-containing neurons in the brain of the white-footed mouse, Peromyscus leucopus

The distribution of immunoreactive LH in the brain of the white-footed mouse (Peromyscus leucopus) was determined using immunocytochemical procedures. Immunoreactive fibers are located in the hypothalamus, preoptic area, septum and amygdala. Stained cell bodies are seen in the arcuate nucleus and preoptic area. Gonadectomy enhances staining for LH in the brain.     

Ecdysteroid receptors located in the central nervous system of an insect

Summary Using thaw-mount autoradiography for steroid hormones, we obtained direct evidence for a nuclear localization of ecdysteroid binding sites in target organs of blowfly (Calliphora vicina) larvae. The binding sites revealed properties of ecdysteroid receptors. Endocrine cells of the ring gland were found to be target tissues of ecydysteroids. This observation provides morphological evidence for a network of complex interendocrine regulation. In the central nervous system receptorcontaining neurons were identified which include many, if not all, neurosecretory cells of the brain. A map of ecdysteroid sensitive cells of the larval brain is presented.     

In vivo inactivation of transglutaminase during the acute acrylamide toxic syndrome in the rat

The activity of liver and brain transglutaminase is rapidly lost following i.p. injection of acrylamide (50-200 mg/kg). Other enzymes investigated were not modified by the treatment, with the exception of brain enolase.     

A convulsant, 3-mercaptopropionic acid,decreases the level of GABA and GAD in rat pancreatic islets and brain

To investigate the properties of the gamma-aminobutyric acid (GABA) synthesizing enzyme, glutamate decarboxylase (GAD), in the brain and the pancreatic islets of the rat, GABA concentration in the brain and the pancreatic islets was measured after intraperitoneal administration of 3-mercaptopropionic acid (3-MP) at 25 mg/kg. 60 min after the administration of 3-MP, GABA concentration in the hypothalamus, the superior colliculus and the hippocampus of the brain decreased by 20–30% and in the pancreatic islets by 35%. The concentration in the pancreatic acini did not change. Western blotting showed that GAD activity in the pancreatic islets decreased after administration of 3-MP compared to the control. The activity of GAD in the pancreatic islets as well as brain can be modified by a convulsant, in this case 3-MP. These results suggest the properties of GAD may be similar in the pancreatic islets and brain.