Differential expression of two neural cell-specific beta-tubulin mRNAs during rat brain development.

Cholecystokinin (CCK) is a neuropeptide found in brain and intestine. In this report, we have isolated a cDNA clone that encodes CCK from a mouse brain cDNA library. This cDNA clone has extensive homology to CCK precursors that have been sequenced previously. Southern blots of genomic DNA probed with this cDNA clone revealed single bands for each of eight different restriction enzymes, all of which could be accounted for by a single genomic clone, suggesting that the CCK gene is present as a single-copy gene in mice. RNA blots, primer extensions, and S1 nuclease protection assays have suggested that the same RNA start site is utilized in brain and in gut. Finally, we have shown, by using RNA blots and a radioimmunoassay specific for CCK, that CCK is expressed at maximum adult levels in intestine at birth but that adult concentrations of CCK and its mRNA are not reached in brain until much later in development.

show abstract

“…SB, upper panel). A similar decrease in the levels of B-tubulin mRNA has recently been reported to occur during rat brain development (32). Fig.…”

Section: Resultssupporting

confidence: 85%

Differential expression of the mouse cholecystokinin gene during brain and gut development.

Friedman¹,

Schneider²,

Powell³

1985

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Sections of experimental L5 DRGs (either NGF-treated or saline-treated) at l-l 2 d after axotomy and sections of normal controls were hvbridized with YS-labeled NF-L cDNA fLewis and Cowan. 1985) or &bulin cDNA (entire clone RBTl; Bond et al, 1984) and washed to a final stringency of 0.5 x SSC with 5 mM DTT at room temperature. On Northern blots of total RNA from rat DRG, the NF-L cDNA hybridizes with 2.5-kb and 4-kb mRNAs, and the @-tubulin cDNA hybridizes with a 1.8-kb mRNA (Wang and Oblinger,199Oa).…”

Section: Methodsmentioning

confidence: 99%

“…Two pg of total RNA were loaded into wells of a slot blot apparatus (American Bionetics) and vacuum filtered onto Nytran membranes (Schleicher & Schuell) according to manufacturer's instructions. The membranes were baked in a vacuum oven at 80°C for 2 hr and then hybridized sequentially to '*P-labeled cDNA probes to NF-L (Lewis and Cowan, 1985) and fl-tubulin (Bond et al, 1984). Final stringency of the washes was 0.1 x SSC with 0.1% SDS at 40°C.…”

Section: Methodsmentioning

confidence: 99%

NGF rescues substance P expression but not neurofilament or tubulin gene expression in axotomized sensory neurons

1991

View full text Add to dashboard Cite

A reduction in the supply of retrogradely transported NGF has been proposed as a possible signal for the axotomy response in dorsal root ganglion (DRG) neurons. Components of the axotomy response that have previously been well characterized in axotomized DRG cells include changes in cytoskeletal gene expression and changes in the expression of neurotransmitters/neuromodulators such as substance P. In this study, we examined the role of NGF in the axotomy response by examining protein synthesis and mRNA levels of the low-MW neurofilament protein (NF-L) and beta-tubulin in DRG cells at 1, 7, and 12 d after axotomy with and without continuous administration of exogenous NGF. We also examined substance P levels in the DRG by immunocytochemistry under the same experimental conditions. Sciatic nerves of adult male rats were unilaterally transected at the midthigh level, and the proximal nerve stumps were placed into Silastic tubes connected to osmotic minipumps that were filled with biologically active NGF. NGF (0.5 mg/ml in saline) was continuously infused (0.5 microliter/hr) onto the proximal stumps of transected sciatic nerves for 1–12 d. Control animals were prepared in an identical fashion except that the nerves were treated with saline alone. At death, DRGs were removed from the animals; the L4 experimental DRGs (axotomized) and contralateral L4 DRGs (uninjured) were used immediately for protein synthesis experiments, while the experimental and contralateral L5 DRGs were fixed in 4% paraformaldehyde and subsequently used for in situ hybridization and immunocytochemistry. From another set of experimental animals, the L4 and L5 DRGs were harvested and used for total RNA isolation and RNA blotting experiments. Immunocytochemical studies using a polyclonal antibody to substance P showed that the immunodetectable levels of this peptide decreased to undetectable levels in DRG neurons after axotomy and saline administration. However, in axotomized neurons treated with NGF, the level of immunodetectable substance P did not decrease, but instead, increased over even that present in normal DRG neurons. Pulse labeling of DRGs with 35S-methionine:cysteine followed by 2-dimensional (2D) gel electrophoresis and fluorography revealed that the synthesis of neurofilament (NF) proteins was decreased, while that of tubulin was increased, 12 d after sciatic nerve transection. NGF administration to axotomized neurons did not alter this pattern. Quantitative analysis of in situ hybridizations of DRG neurons and RNA blot analysis with cDNA probes specific for NF-L and beta-tubulin mRNAs showed that NGF treatment of axotomized DRGs did not significantly affect cytoskeletal gene expression at the mRNA level.(ABSTRACT TRUNCATED AT 400 WORDS)

show abstract

“…The intensity of the TM mRNA band rapidly decreased as a function of time of incubation with monokine, whereas the size of the TM mRNA remained unchanged at 3.8 kilobases. We excluded the possibility that TNF was causing a nonspecific reduction in total cellular mRNA by demonstrating that the cytokine had no suppressive effect on the concentrations of tubulin mRNA (2) (Fig. 2A) or the levels of glyceraldehyde-3-phosphate-dehydrogenase mRNA (6) and angiotensinogen mRNA (20) (data not shown).…”

mentioning

confidence: 99%

“…The bands were transferred to a GeneScreen filter (Du Pont, NEN Research Products, Boston, Mass.) by capillary blotting, and the filter was hybridized with random primer 32P-labeled DNA probes (8) for TM (12,13) and rat brain P-tubulin (2). Transcription of the ,-tubulin gene is known to produce two separate mRNAs.…”

mentioning

confidence: 99%

Tumor necrosis factor suppresses transcription of the thrombomodulin gene in endothelial cells.

Conway¹,

Rosenberg²

1988

Mol. Cell. Biol.

332

176

View full text Add to dashboard Cite

Tumor necrosis factor (TNF) dramatically alters the levels of various surface components of the blood vessel wall, such as blood coagulation enzyme receptors, leukocyte-adhesive receptors, and class 1 major histocompatibility complex antigens, which may have relevance to its effects in septic shock, angiogenesis, and tumor growth. However, the precise mechanism by which the cytokine is able to accomplish this remodeling of the endothelial cell surface has not been defined. We have demonstrated that exposure of bovine and human endothelial cells to TNF leads to suppression of the functional cell surface thrombin receptor, thrombomodulin (TM), and TM mRNA of virtually identical magnitude. The cytokine has no significant effect on the stability of TM mRNA or endothelial receptor turnover. Nuclear run-on studies reveal that the treatment of endothelial cells with TNF for short periods reduces TM gene transcription to as little as 3% of control values and that this inhibition does not require new protein synthesis.

show abstract

Differential expression of two neural cell-specific beta-tubulin mRNAs during rat brain development.

Cited by 96 publications

References 30 publications

Differential expression of the mouse cholecystokinin gene during brain and gut development.

Differential expression of the mouse cholecystokinin gene during brain and gut development.

NGF rescues substance P expression but not neurofilament or tubulin gene expression in axotomized sensory neurons

Tumor necrosis factor suppresses transcription of the thrombomodulin gene in endothelial cells.

Contact Info

Product

Resources

About