Play Doh\u27s Cave and The Pursuit of the American Cream

Take a minute. Imagine Wiley Coyote and Road Runner are in a domestic partnership. What would that look like? Close your eyes and Pause for 30 seconds. Don’t you see? Coyote never catches up. They keep running faster and faster. Everything in the house gets swept into the whirlwind they’ve created in their paths - the books, the shelves, the bed, and the desk lamp. Their circling movement creates a vacuum, which ultimately causes the entire structure to implode upon itself. This text is an examination of my work and its relationship to the economic and the domestic. The metaphor of the tragicomic perpetually failing in the spotlight is a dominant motif standing against a backdrop of an overflowing bloat of unidentifiable mass desperately trying to repel gravity. In the first section of this text I offer a brief overview of my two-year trajectory, and an analytical perspective of my culminating thesis exhibition. In the second section, I share with you a trough of incomplete jokes, and standalone punch lines. This Rolodex I keep of “word sketches” catalogues my search for the shape of a laugh

The Biosynthesis of Transfer Ribonucleic Acid in the Developing Rat Brain and in Cultured Glial Cells

The biosynthesis of tRNA was investigated in cultured astroglial cells and the 3-day-old rat brain in vivo . In the culture system astrocytes were grown for 19 days and were then exposed to [ 3 H]guanosine for 1.5–7.5 h; 3-day-old rats were injected with [ 3 H]guanosine and were killed 5–45 min later. [ 3 H]tRNA was extracted, partially purified, and hydrolyzed to yield [ 3 H]-guanine and [ 3 H]methyl guanines. The latter were separated from the former by high performance liquid chromatography and their radioactivity determined as a function of the time of exposure to [ 3 H]guanosine. The findings indicate that labeling of astrocyte tRNA continued for 7.5 h and was maximal, relative to total RNA labeling, at 3 h, while in the immature brain tRNAs were maximally labeled at 20 min after [ 3 H]guanosine administration. The labeling pattern of the individual methyl guanines differed considerably between astrocyte and brain tRNAs. Thus, [ 3 H]1-methylguanine represented up to 35% of the total [ 3 H]methyl guanine radioactivity in astrocyte [ 3 H]tRNA, while it became only negligibly labeled in brain [ 3 H]tRNA. Conversely, brain [ 3 H]tRNA contained more [ 3 H]N 2 -methylguanine than did astrocyte [ 3 H]tRNA. Approximately equal proportions of [ 3 H]7-methylguanine were found in the [ 3 H]tRNAs of both neural systems. The [ 3 H]methylguanine composition of brain [ 3 H]tRNA was followed through several stages of tRNA purification, including benzoylated DEAE-cellulose and reverse phase chromatography (RPC-5), and differences were found between the [ 3 H]methylguanine composition of RPC-5 fractions containing, respectively, tRNA lys and tRNA phe . The overall results of this study suggest that developing brain cells biosynthesize their particular complement of tRNAs actively and in a cell-specific manner, as attested by the significant differences in the labeling rates of their methylated guanines. The notion is advanced that cell-specific tRNA modifications may be a prerequisite for the successful synthesis of cell-specific neural proteins.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65406/1/j.1471-4159.1980.tb09020.x.pd

A REGIONAL STUDY OF SOME OSMOTIC, IONIC AND AGE FACTORS AFFECTING THE STABILITY OF CEREBRAL LYSOSOMES 1

An examination was made of the effect of changes in the osmolarity and the ionic composition of the homogenizing medium on the partition of lysosomal arylsulphatase and N -acetylglucosaminidase of cerebral cortex, hypothalamus and thalamus of the rat. Sulphatase appeared to be more sensitive to hypotonicity than glucosaminidase, since a higher proportion of the sulphatase was released from the lysosomes into the soluble fraction of the cells from all three neuroanatomical areas examined. In the presence of 250 mM-sucrose, supplementation with 10 mM-Mg led to clumping of the lysosomes and their translocation into the heavy-particulate fraction; no such effect of 10 mM-Mg was noted in the absence of 250 mM-sucrose. The intracellular distribution of bound N -acetylneuraminic acid (bound-NANA) was also examined. The shifts observed in its intracellular localization as a result of changes in the ionic composition of the homogenizing medium rule out bound-NANA as a structural component of the membrane of the cortical lysosome. However regional differences in the response of bound-NANA to ionic factors were observed. Lysosomes from cerebral cortex of adult and 12-day-old rats were also compared. Differences in the pattern of distribution of lysosomes in linear sucrose gradients and in response to ionic factors were uncovered. The results support the previously enunciated concept (Sellinger and Hiatt, 1968) of a regional microheterogeneity of lysosomes and add a new, age-related dimension to it.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65807/1/j.1471-4159.1969.tb05969.x.pd

PROTEIN SYNTHESIS IN NEURONS AND GLIAL CELLS OF THE DEVELOPING RAT BRAIN: AN IN VIVO STUDY 1

A technique for the isolation of pure neuronal perikarya and intact glial cells from cerebral cortex has been developed for routine use. The yield of neuronal perikarya and glial cells was greater from highly immature (5–10 days) rat cerebral cortex than from the cortex of older rats (18–43 days). The perikarya/glia yield ratio decreased with age indicating that, as the glial population matured, the procedure succeeded in isolating a gradually smaller proportion of the existing neurons. The perikarya/glia ratio was highest for the 5-day-old cortex in which no mature glial cells could be identified. After a 10-min pulse in vivo of intrathecally injected [ 14 C]phenylalanine, the specific radioactivity of the neuronal proteins was higher than that of the glial proteins in the 5-, 10- and 18-day-old rat but was lower in the 43-day-old rat. The values for absolute specific radioactivity of the 14 C-labelled proteins in both cell types were greater, the younger the brain. The 14 C-labelling of neuronal and glial proteins in the 18-day-old rat was assessed in vivo as a function of time by determining the incorporation of [ 14 C]phenylalanine into such proteins at 5, 10, 20 and 45 min after administration of the amino acid. The rate of incorporation of [ 14 C]phenylalanine into the glial cells was faster than into the neurons since higher specific radioactivities of the glial proteins could be achieved at earlier times. Also, a biphasic pattern of 14 C-labelling of the glial proteins was noted, suggesting, perhaps, a sequential involvement of the oligodendrocytes and astrocytes. Homogenates of prelabelled neuronal perikarya were fractionated into the nuclear, mitochondrial microsomal and soluble cell sap fractions. In the 18-day-old cerebral cortex, the proteins of the microsomal fraction exhibited the highest specific radioactivity at the end of 10 min, whereas by 20 min proteins of the mitochondrial fraction were most highly labelled. The specific radioactivity of the nuclear proteins increased over the entire 45-min experimental period. On the contrary, the proteins of the soluble cell sap, in which the specific radioactivity was at all times by far the lowest, were maximally labelled by 5 min. Examination of the labelling of the neuronal subcellular fractions as a function of age revealed that at 10 min after administration of [ 14 C]phenylalanine, the specific radioactivities of all 14 C-labelled proteins were highest in the youngest (5-day-old) neurons. The proteins of the microsomal fraction were most rapidly labelled at all ages. During this interval the proteins of the soluble cell sap were only moderately labelled in the 5-day-old neurons and were totally unlabelled in the 43-day-old neurons, indicating age-dependent differences in the rate of utilization of the amino acid precursor by the neurons.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65968/1/j.1471-4159.1971.tb00007.x.pd

METHYLATION OF E. COLI TRANSFER RIBONUCLEIC ACIDS BY A tRNA ADENINE-l-METHYLTRANSFERASE FROM RAT BRAIN CORTEX AND BULK-ISOLATED NEURONS

Brain cortices or bulk-isolated neuronal cell bodies prepared from cortices of 8-day old male rats were used as the source of a l-methyl adenine-specific tRNA methyltransferase (tRNA-AMT). Ammonium sulfate fractionation and chromatography on spheroidal hydroxylapatite and Sephadex G-200 yielded an 80-fold purified enzyme, as determined by using E. coli bulk tRNA as substrate. The kinetic parameters of tRNA-AMT for the substrate S -adenosyl-l-methionine (SAM) ( K m = 6 ΜM) and the inhibitor, S -adenosyl-l-homocysteine (SAH) ( K i = 3.4 Μm) were determined and several SAH analogs tested as inhibitors. S -Adenosyl-l-cysteine (SAC) ( 10 -4 m) and S -adenosyl-d-homocysteine (SADH) (10 -4 m) produced a 35 and a 21% reduction in enzyme activity, respectively. The effects of Mg 2+ , NH 4 + acetate and of the polyamines spermine, putrescine and spermidine on the brain tRNA-AMT mimicked the effects of these agents on hepatic tRNA-AMT (Glick et al , 1975). Comparing the ability of cerebral tRNA-AMT to methylate E. coli tRNA glu2 , tRNA val , tRNA phe and bulk tRNA revealed tRNA glu2 as the best and tRNA phe as the least effective substrate. tRNA-AMT prepared from neuronal cell bodies showed closely similar characteristics to the cortical enzyme. A comparison of the activities of tRNA-AMT in neurons and glial cells revealed higher values in the former.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66287/1/j.1471-4159.1978.tb12436.x.pd

THE ELEVATION OF CEREBRAL HISTAMINE- N -AND CATECHOL- O -METHYL TRANSFERASE ACTIVITIES BY l-METHIONINE-dl- SULFOXIMINE 1

The administration of the convulsant, l-methionine-dl-sulfoximine (MSO), increased histamine N -methyl transferase (E.C. 2.1.1.8) (HMT) activity in rat and mouse brain and, to a lesser extent, catechol- O -methyl transferase (E.C. 2.1.1.6) (COMT) activity in rat brain. The duration of this effect was shortened by co-administration of l-methionine. The increased HMT activity was seen in 5 or 7 rat brain regions tested. l-Methionine administration had no effect on the activity of either enzyme. Partially purified HMT preparations from rat or guinea-pig brain exhibited no alterations in activity after the in vitro addition of MSO or l-methionine over a wide range of histamine and S -adenosyl-l-methionine concentrations. Rat brain COMT was equally unaffected by MSO and l-methionine. The in vitro inhibition of HMT and COMT by S -adenosyl-l-homocysteine was the same whether tested on preparations derived from MSO-treated or control animals. The data are discussed with respect to the possible involvement of aberrant methylation processes in the MSO-induced seizure.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65751/1/j.1471-4159.1975.tb07696.x.pd