Biomedical subjects

H Ming

Publications and source records attributed to H Ming.

4 recordsLinked to original sources

PubMed2000

A novel system A isoform mediating Na+/neutral amino acid cotransport.

A cDNA clone encoding a plasma membrane alanine-preferring transporter (SAT2) has been isolated from glutamatergic neurons in culture and represents the second member of the system A family of neutral amino acid transporters. SAT2 displays a widespread distribution and is expressed in most tissues, including heart, adrenal gland, skeletal muscle, stomach, fat, brain, spinal cord, colon, and lung, with lower levels detected in spleen. No signal is detected in liver or testis. In the central nervous system, SAT2 is expressed in neurons. SAT2 is significantly up-regulated during differentiation of cerebellar granule cells and is absent from astrocytes in primary culture. The functional properties of SAT2, examined using transfected fibroblasts and in cRNA-injected voltage-clamped Xenopus oocytes, show that small aliphatic neutral amino acids are preferred substrates and that transport is voltage- and Na(+)-dependent (1:1 stoichiometry), pH-sensitive, and inhibited by alpha-(methylamino)isobutyric acid (MeAIB), a specific inhibitor of system A. Kinetic analyses of alanine and MeAIB uptake by SAT2 are saturable, with Michaelis constants (K(m)) of 200-500 microm. In addition to its ubiquitous role as a substrate for oxidative metabolism and a major vehicle of nitrogen transport, SAT2 may provide alanine to function as the amino group donor to alpha-ketoglutarate to provide an alternative source for neurotransmitter synthesis in glutamatergic neurons.

Amino Acid Sequence↗

PubMed2000

Cloning and functional identification of a neuronal glutamine transporter.

Glutamine is the preferred precursor for the neurotransmitter pool of glutamate, the major excitatory transmitter in the mammalian central nervous system. We have isolated a complementary DNA clone (designated GlnT) encoding a plasma membrane glutamine transporter from glutamatergic neurons in culture, and its properties have been examined using the T7 vaccinia system in fibroblasts. When GlnT is transfected into CV-1 cells, L-glutamine is the preferred substrate. Transport is Na(+)-dependent and inhibited by alpha-methylaminoisobutyric acid, a specific inhibitor of neutral amino acid transport system A. Kinetic analysis of glutamine uptake by GlnT is saturable, with a Michaelis constant (K(m)) of 489 +/- 88 microM at pH 7.4. Glutamine uptake mediated by GlnT is pH-sensitive with a 5-fold greater efficiency of uptake at pH 8.2 than at pH 6.6. Only the maximal velocity of transport increases without a significant change in K(m). The distribution of GlnT mRNA and protein in the central nervous system is widespread and is expressed on neurons that use glutamate as their neurotransmitter. In cultured cerebellar granule cells, GlnT is expressed only on neurons and is absent from astrocytes. GlnT expression increases concomitantly with the morphologic and functional differentiation of these cells in vitro, consistent with its role of supplying glutamatergic neurons with their neurotransmitter precursor. GlnT is the first member of the system A family of neutral amino acid transporters with 11 putative membrane-spanning domains and is a potential target to modulate presynaptic glutamatergic function.

Amino Acid Transport Systems, Basic↗

PubMed2000

Differential actions of neurotrophins on apoptosis mediated by the low affinity neurotrophin receptor p75NTR in immortalised neuronal cell lines.

The low affinity neurotrophin receptor (p75NTR) mediates apoptosis of a number of neuronal and non-neuronal cells but the signals leading to the apoptosis remain obscure. To reveal the mechanism of p75NTR-mediated apoptosis, a neural cell line expressing human p75NTR was established. The human cDNA fragment encoding for p75NTR was PCR-amplified, cloned into the retrovirus expression vector pXT-1 and transfected into the rat cerebellum cell line R2. The expression of p75NTR in the R2 cell line was demonstrated by both Northern blotting analysis and immunocytochemistry. Serum withdrawal induced dramatic apoptosis in p75NTR-expressing R2 cells (R2L1) but not in pXT-1 transfected control R2 cells (R2P). Reverse transcription polymerase chain reaction (RT-PCR) revealed that these cell lines express trkA and trkB but not trkC. The apoptosis of R2L1 cells triggered by the serum deprivation for 48 h was completely prevented by neurotrophin-3 and the antibody to p75NTR but only partially prevented by the nerve growth factor and brain derived neurotrophic factor. We conclude that the p75NTR mediates apoptosis of R2L1 cells by its intrinsic receptor effects requiring an unbound status of this receptor and that the apoptosis is prevented by neurotrophins or the antibody to p75NTR through distinct mechanisms.

Animals↗

PubMedDate not supplied

[Macrocheilia].

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Adult↗