Leonhard S. Wolfe

High Levels of Brain Dolichols in Neuronal Ceroid-Lipofuscinosis and Senescence

Abstract: Dolichols as unesterified alcohols were identified as significant components of lipid extracts from storage cytosomes isolated post‐mortem from the brains of patients with the infantile, late infantile, and juvenile types of neuronal ceroid‐lipofuscinosis (NCL). Very small amounts of dolichols were present in the corresponding subcellular fractions of non‐NCL brains. The nuclear fraction from NCL cerebral cortex contained the highest dolichol content expressed per milligram protein or lipid, whereas the crude mitochondrial fraction was the richest in normal brain. Highly significant elevations of dolichol levels were found in human cerebral cortex of patients with NCL and Alzheimers disease compared with age‐matched controls, but the levels were normal in Picks disease. In human non‐NCL cerebral cortex, dolichols increased from 16 μg/g at age 5 to over 200 at age 81. Rat cerebral cortex showed a similar progressive increase in dolichol content with age. The high dolichol values in NCL, Alzheimers disease, and senescence appears to be related to the increase of lipofuscin in brain. This is the first time a uniform biochemical abnormality has been found in all childhood forms of NCL, but the enzyme defect is still unidentified. It may lie on pathways where dolichols and retinyl compounds are recycled in Golgi membranes and derived organelles during the biosynthesis of glycoproteins.

Dolichols in brain and urinary sediment in neuronad ceroid lipofuscinosis

long-chain polyisoprenoid alcohols (dolichols) increase more than tenfold from age 5 to 80 years in human cerebral cortex. The dolichol content of brain from infantile, late infantile, and juvenile forms of neuronal ceroid Lipofuscinosis (NCL) was significantly higher than in age-matched patients with other neurologic diseases. Significant increase of dolichols was also found in the urinary sediment in all three types of NCL patients, and this test is useful in making the diagnosis. Dolichol accumulation is the first biochemical marker of NCL and seems to parallel storage of ceroid lipofuscin.

Possible Roles of Prostaglandins in the Nervous System

Few would question that one of the most exciting and important developments in biochemistry and physiology in recent years has been the emergence of prostaglandins as a new class of molecules mediating and modulating hormonal, neurohormonal, or other stimuli on most mammalian tissues. In just over ten years the literature on prostaglandins has expanded to over 5000 references, from basic chemistry, metabolism, and pharmacology to clinical trials of natural prostaglandins and synthetic analogues. Although the subject is recent, one can look back at the work of others over the 30 years before the announcement of the structures of the six primary prostaglandins by Bergstrom and coworkers (Bergstrom et al., 1962, 1963; review, 1968) and identify principles with biological activities that we now know are mainly due to prostaglandins. Foremost among these were the independent observations of Kurzrok and Lieb (), Goldblatt (1933, 1935), and von Euler (1934, 1935a) that human semen and extracts of sheep vesicular glands contained factors that stimulated intestinal and uterine muscle and lowered arteral blood prossure. Von Euler (1935b) showed that these activities were due to acidic lidic lipids and proposed the name “prostaglandin”.

A type of juvenile cerebromacular degeneration characterized by granular osmiophilic deposits

Abstract An 8-year-old girl developed a progressive neurological disease, characterized by minor seizures, gradual loss of vision leading to complete blindness, and ultimate dementia. She died at 16 years of age. At post-mortem virtually all neurons contained insoluble, autofluorescent, SBB-positive deposits. Similar deposits were present in many viscera. They were particularly abundant in skeletal muscle. By electron microscopy they appeared as granular osmiophilic deposits (GROD). The epithelial cells of the renal glomeruli contained lamellar inclusions in addition to the GROD. GROD and lipofuscin share many histochemical and ultrastructural features, but minor differences suggest that their chemical composition may differ. Chemical analysis of tissue from the patient failed to identify the nature of the GROD, principally because of their insolubility. This patient clinically conforms to the syndrome of juvenile cerebromacular degeneration. In these patients histochemical and ultrastructural examination of skin and skeletal muscle seems to be the most convenient method of making a precise pathological diagnosis at present.

Adult onset motor neuronopathy in the juvenile type of hexosaminidase A and B deficiency.

Two sisters presented with progressive muscle cramps, as well as wasting and weakness of the legs with onset after age 20. They also showed intention tremor of the upper extremities and dysarthria starting during the first decade. The older patient also had fasciculations; the younger, hyperreflexia. Total plasma beta-hexosaminidase (Hex) activity with 4-methylumbelliferyl-acetyl-glucosamine as substrate was reduced to 1.4% and 2.7% of the control in the 2 patients, respectively. Hex A activity measured by 4-methylumbelliferyl-N-acetylglucosamine-6-O-sulphate as substrate was 9.9% and 12.8% of the mean control value in the 2 patients, respectively. Hex B activity was undetectable in both patients. Leukocyte total Hex activity was 7-8% of normal; residual Hex A activity in the 2 patients was 17.8% and 16.3% of normal controls, respectively. Fibroblastic residual Hex A activity in the 2 patients was 9.6% and 22% of normal mean value, respectively. Appendiceal ganglion cells contained membranous cytoplasmic bodies in the younger patient. Thin layer chromatography of the appendiceal extract from one patient (III/2) showed a marked increase of GM2 ganglioside, and some increase of GM3 ganglioside. Northern blots performed on fibroblast cell lines from both patients for the demonstration of alpha and beta locus messenger RNA showed no difference between patients and control. These patients have a rare form of adult-onset progressive motor neuron disease presumably due to abnormal beta subunits, causing severe deficiency of both Hex A and Hex B. The phenotypic expression of this disease is similar to motor neuron disease due to alpha locus mutations, which suggests that the Hex A deficiency, even though only a partial one, may be the important pathogenic factor.

Effect of Drugs on Local Cerebral Glucose Utilization in Traumatized Brain: Mechanisms of Action of Steroids Revisited

It may seem inappropriate to include in this volume work which does not deal directly with cerebral edema. However, we make no apologies for this fact since our present studies on the mechanisms by which injury to the brain causes functional neurological disturbances have evolved from a longstanding interest of one of us in processes underlying the development of brain edema and its consequences. We believe that the implications of our findings are pertinent to consideration of any condition associated with functional disturbances which are generally, rightly or wrongly, ascribed to cerebral edema.

Infantile neuronal ceroid lipofuscinosis: Isolation of storage material

We used ultracentrifugation to isolate autofluorescent storage particles from the brains of patients with infantile type of neuronal ceroid lipofuscinosis (Haltia-Santavuori). The particles differed from those of the late infantile (Jansky-Bielschowsky) and juvenile (Batten-Spielmeyer-Sjögren) types and retained the characteristic appearance—large, partly membrane-bound conglomerates of spherical globules with a uniform finely granular internal structure—despite the vigorous procedure. The sedimentation properties of the particles also differed from the two other forms, but dolichols are found in the particles of all three diseases.

Neurochemical Sequelae of Brain Damage and Their Role in Functional Disturbances

Various types of injury have been shown to produce a variety of chemical changes in brain which are thought to underly functional disturbances resulting from the particular insult. Some of the systems known to be affected in injured brain are summarized in Figure 1, but the list is by no means exhaustive. The exact mechanisms by which the chemical perturbations are related to functional disturbances are not understood in most cases and it is often difficult to distinguish between the cause and the effect in this relationship. One problem has been the difficulty in assessing neurological function in animals since normalization of neurochemical disturbances in brain cannot always be equated with normally functioning central nervous system and the available methods are not necessarily applicable in the particular experimental situation.

The Biosynthesis of Prostaglandins and Thromboxanes by Nervous Tissue

Prostaglandins F2α and E2 are rapidly synthesized and released in vivo from various regions of the central nervous system and from autonomic nerves during stimulation or after trauma (Wolfe, 1975). They alter the effects of neurotransmitters at pre- and postsynaptic sites. PG’s appear in increased amounts in human CSF following epileptic seizures, trauma, infections and stroke (Wolfe & Mamer, 1975). Measurement of PG biosynthesis by cerebral cortex and other brain regions in vitro shows considerable biosynthetic capacity from endogenous precursors, Catabolic enzyme activities however are exceedingly low. A metabolically stable pool of arachidonic acid appears in brain tissue after animal death which saturates the brain fatty acid cyclo-oxygenase (Wolfe et al., 1976a; 1976c) localized in microsomal membrane elements (Baker, R. R. & Wolfe, L. S., unpublished results).

Biologically Active Lipids

For many years it has been realized that tissues and body fluids, including the brain and cerebrospinal fluid, contain pharmacologically active substances that have the properties of acidic lipids.(1–7) These substances are most often detected by their ability to elicit contractions of a slow type in smooth muscle that are not affected by atropine or antagonists of 5-hydroxytrypt-amine and histamine. Their activity is not diminished by preincubation with crystalline chymotrypsin, trypsin, or the broad-spectrum protease from Streptomyces griseus, Pronase. Most substances in this class partition into solvents such as diethyl ether, ethyl acetate, or chloroform from acidic but not basic aqueous solutions or extracts. Thus they behave as carboxylic acids. Chemical treatment by carboxyl-complexing reagents (e.g., N,N′-carbo-di-p-tolylimide), hydroxyl-complexing reagents (e.g., phenylisocyanate), and procedures which react with double bonds (e.g., hydrogenation, bromination) abolishes in most cases the smooth-muscle contracting property.(8) These simple procedures suggest compounds which are unsaturated hydroxy fatty acids. Recent investigations have shown that prostaglandins are by far the most important group of physiologically and pharmacologically active substances of this class. Particular members of this new and structurally unique lipid class are among the most powerful smooth-muscle contracting substances known and occur in varying amounts in most if not all tissues.