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Biomedical subjects

D Zelenika

Publications and source records attributed to D Zelenika.

3 recordsLinked to original sources

A novel transcript overlapping the myelin basic protein gene.

Myelin basic protein (MBP) is a major constituent of myelin synthesized by oligodendrocytes and Schwann cells. We have investigated the expression of mouse MBP RNAs outside the nervous system. Nuclease protection experiments indicate that RNAs containing exon 1 and not the six downstream exons of the MBP gene are transcribed in various hemopoietic tissues. We have isolated a hemopoietic MBP-related (HMBPR) cDNA clone from a mouse bone marrow cDNA library screened with an MBP cDNA probe. This clone contains exons 1a and 1b and a part of intron 1 of the MBP gene. An additional 5' region, encoded by at least three unidentified exons, lies upstream of exon 1a. The HMBPR clone corresponds to a 5-kb RNA expressed in bone marrow, spleen, thymus, and macrophagic cells. This transcript is expressed at a similar level in brain, although at a lower level than the classical 2-kb mRNA. These data indicate that a new transcript, overlapping the MBP transcription unit and controlled by a distinct promoter, is expressed in hemopoietic tissues. This RNA might encode a 21-kDa protein sharing a common domain with MBP.

Amino Acid Sequence

Isolation of a novel cDNA corresponding to a transcript expressed in the choroid plexus and leptomeninges.

The CNS is composed of neurons and glial cells (i.e., astrocytes, oligodendrocytes, and microglia). The brain communicates with the blood circulation through choroid plexus and meninges as well as through the blood-brain barrier. To identify transcripts specifically expressed in a distinct brain cell type, we have previously constructed a subtracted cDNA library from the poly(A)+ RNAs of a velate protoplasmic-like astrocytic cell line, designated D19. This library was screened in order to isolate transcripts over-expressed in this astroglial cell line versus another astroglial cell line. Of the six recombinants which have been isolated, three sequences have not been described. Their sizes ranged from 100 to 200 bp and they hybridized to mRNAs expressed in vivo inside and outside the CNS. We have constructed a size-selected D19 cDNA library in order to obtain the full length cDNAs corresponding to the three undescribed sequences. We report here the isolation of a 1.6 kb cDNA corresponding to one of these recombinants, named p14. Its sequence has not yet been described and its deduced amino acid sequence codes for a 43 kDa protein with a putative signal peptide. In situ hybridization shows that this transcript is expressed at a high level in choroid plexus and leptomeninges and also in perivascular cells in the adult mouse brain. It is also expressed in cell subsets of kidney and gonads as well as in perivascular cells in skeletal and cardiac muscles. These localizations suggest that the encoded protein might be involved in transport processes and hormonally controlled.

Animals

[Membrane abnormalities and cellular hyperreactivity in different models of hypertension].

In various models of hypertension of genetic origin, a hypersensitivity of phospholipase C has been demonstrated to participate in the hyperreactivity of platelets toward a variety of vasoactive agents. Since this abnormality could not be observed in the absence of cell stimulation, it could not account for the increase in free Ca2+ which has been reported in resting platelets in primary hypertension. Likewise, in hypertensive subjects, platelets behave hyperactive when stimulated by ADP, although the stimulus has been demonstrated to be a poor activator of phospholipase C. In order to gain insight into the membrane alteration that could account for the cellular hyperactivity which characterizes hypertensive subjects, we investigated, in resting platelets, the kinetics of radioactive labeling of major membrane phospholipids. Isolated platelets were prepared from SHR (4w and 17w of age), SHR-SP, Dahl salt-resistant and salt-sensitive rats fed either a low or a high salt diet, DOCA-salt hypertensive rats and from the appropriate normotensive controls. Irrespective of the radioactive precursor used (32P-orthophosphate, 3H-glycerol, 3H-choline), the labeling of phosphatidylcholine (PC) was markedly (up to 20 fold) enhanced in SHR (whichever their age) and SHR-SP compared with WKY. This increase, specific of PC, could not be accounted for by differences either in the actual amount of PC or in the uptake of various labels, suggesting an increased PC turnover. Such an increase was also observed in platelets of Dahl hypertensive rats but not in those of DOCA-salt hypertensive rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals