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PubMed1997

Visual sensitivities of nur77 (NGFI-B) and zif268 (NGFI-A) induction in the suprachiasmatic nucleus are dissociated from c-fos induction and behavioral phase-shifting responses.

Mammalian circadian rhythms are regulated by a pacemaker in the suprachiasmatic nucleus of the hypothalamus. Recent work from several laboratories has shown that light induces the IEGs, c-fos and jun-B, in the rodent suprachiasmatic nucleus. In hamsters, there is a strong correlation between circadian entrainment and the induction of c-fos and jun-B in the suprachiasmatic nucleus by light. Previous work has shown that the IEGs, nur77 and zif268, both of which encode transcription factors, are also light-inducible in the rat suprachiasmatic nucleus [Rusak, B., McNaughton, L., Robertson, H.A. and Hunt, S.P., Circadian variation in photic regulation of IEG mRNAs in rat suprachiasmatic nucleus cells, Mol. Brain Res., 14 (1992) 124-130.; Sutin, E.L. and Kilduff, T.S., Circadian and light-induced expression of IEG mRNAs in the rat suprachiasmatic nucleus, Mol. Brain Res., 15 (1992) 281-290.]. To characterize the photic-regulation of these genes in the suprachiasmatic nucleus of golden hamsters, we used in situ hybridization to measure nur77 and zif268 mRNA levels with 33P-labeled complementary RNA probes. 5-min monochromatic light pulses at CT19 induced a dramatic increase in both nur77 and zif268 mRNA levels. Peak mRNA levels occurred 45-60 min after light onset for both nur77 and zif268. In addition, the induction of both nur77 and zif268 mRNA levels was gated by the circadian pacemaker. Light pulses during subjective day (CT3 and CT9), which do not cause behavioral phase-shifts, did not significantly alter mRNA levels of either nur77 or zif268; whereas light pulses during the subjective night (CT14 and CT19), which induce phase-shifts, dramatically increased both nur77 and zif268 mRNA levels. In contrast to c-fos induction, which has a photic threshold indistinguishable from that of the behavioral phase-shifting response, nur77 and zif268 mRNA induction were found to have visual sensitivities greater than the phase-shifting response by 1-2 log units (10-100-fold). Although light and circadian phase regulate nur77 and zif268 expression in the SCN, these results demonstrate that their induction is not rate-limiting for photic entrainment of the hamster circadian system.

Animals↗
PubMed1998

Precise distribution of neuronal nitric oxide synthase mRNA in the rat brain revealed by non-radioisotopic in situ hybridization.

Regional distribution of neurons expressing neuronal nitric oxide synthase mRNA in the rat brain was examined by non-radioisotopic in situ hybridization, using digoxigenin-labeled complementary RNA probes. Clustering of intensely positive neurons was observed in discrete areas including the main and accessory olfactory bulbs, the islands of Calleja, the amygdala, the paraventricular nucleus of the thalamus, several hypothalamic nuclei, the lateral geniculate nucleus, the magnocellular nucleus of the posterior commissure, the superior and inferior colliculi, the laterodorsal and pedunculopontine tegmental nuclei, the nucleus of the trapezoid body, the nucleus of the solitary tract and the cerebellum. Strongly-stained isolated neurons were scattered mainly in the cerebral cortex, the basal ganglia and the brain stem, especially the medulla reticular formation. In the hippocampus, an almost uniform distribution of moderately stained neurons was observed in the granular cell layer of the dentate gyrus and in the pyramidal cell layer of the Ammon's horn, while more intensely stained isolated neurons were scattered over the entire hippocampal region. These observations can serve as a good basis for studies on function and gene regulation of neuronal nitric oxide synthase.

Animals↗
PubMed1998

Discrete distribution of the neuropeptide Y Y5 receptor gene in the human brain: an in situ hybridization study.

The present study investigated the regional distribution of putative 'food-intake'-related neuropeptide Y Y5 receptor gene using cRNA in situ hybridization in various regions of the normal control post-mortem human brain. Interestingly, significant levels of Y5 receptor expression were detected in the hypothalamus; the arcuate nucleus being particularly enriched compared to other hypothalamic nuclei. Surprisingly, strong hybridization signals were also noted in the stratum granulosum of the dentate gyrus contrasting with lower levels of Y5 receptor transcripts in other regions of the hippocampal formation. The cerebral cortex, basal ganglia and thalamus were not enriched with Y5 receptor mRNA. It thus appears that the expression of the Y5 receptor gene in the human brain is rather restricted with enrichment in areas consistent with the involvement of this receptor type in the modulation of appetite and seizures.

Aged↗
PubMed2003

Cellular delivery of peptide nucleic acid (PNA).

Peptide nucleic acid (PNA) is a DNA mimic having a pseudopeptide backbone that makes it extremely stable in biological fluids. PNA binds complementary RNA and DNA with high affinity and specificity. These qualities make PNA a leading agent among "third generation" antisense and antigene agents. Unfortunately, fast progress in the exploration of PNA as an experimental and therapeutical regulator of gene expression has been hampered by the poor cellular uptake of PNA. However, a number of transfection protocols for PNA have now been established. These include microinjection, electroporation, co-transfection with DNA, conjugation to lipophilic moieties, conjugation to peptides, etc. Here we give a short introduction to the basic findings on PNA as an antisense and antigene agent in cell-free in vitro systems. This is followed by a comprehensive evaluation of the most interesting literature concerning cellular delivery and the intracellular effect of PNA. Also the current progress as regards using PNA as co-factor in DNA delivery is reviewed.

Animals↗
PubMed1994

Alterations in the expression of osteonectin, osteopontin and osteocalcin mRNAs during the development of skeletal tissues in vivo.

Heterogeneity in the expression of three members of non-collagenous matrix proteins in osteogenic and chondrogenic development in vivo were investigated by in situ hybridization. Sections of several skeletal tissues from mice at various stages of development were hybridized with digoxigenin-labeled complementary RNA probes encoding osteonectin (Osn), osteopontin (Osp) and osteocalcin (Osc). In calvariae and mandibulae, Osn messenger RNA (mRNA) was detected in cells in pre-osseous and osseous tissues before mineralization. Osp mRNA was found in cells attached to the mineralized bone matrix together with Osn mRNA followed by the expression Osc mRNA. In long bones, mRNAs for Osn, Osp and Osc were sequentially expressed with bone development from primary spongiosa to diaphyseal bone. In growth cartilage, Osn mRNA was observed in chondrocytes in non-mineralized cartilage, whereas Osp mRNA was detected in hypertrophic chondrocytes in mineralized cartilage matrix with a characteristic switch in expression. Osc mRNA was not detected in any chondrocytes. These results indicate that osteogenic differentiation in bone development in vivo is characterized by the sequential expression of these three genes, and suggest that these genes are expressed differentially and specifically, in association with extra-cellular matrix mineralization.

Animals↗
PubMed2002

Sensory nerves and neuropeptides in uterine cervical ripening.

At the time of parturition (fetal delivery) the uterine cervix must "ripen," becoming soft, pliable, and dilated to accommodate the fetus' delivery. The fundamental processes underlying cervical ripening remain poorly understood. Knowledge that abundant autonomic and sensory nerves supply the uterine cervix, that transection of afferent nerves supplying the cervix blocks parturition, and that some of the changes in the cervix resemble those seen in inflammatory reactions suggests nerves may have a role in the cervical ripening changes. The present study utilized immunohistochemistry, plasma extravasation, and solution hybridization-nuclease protection assay to elucidate the complement of primary afferent nerves and some receptors in the rat cervix during pregnancy, and to determine if they may have roles in the ripening process at term. This study revealed an abundance of nerves associated with the cervical vasculature and myometrial smooth muscle containing immunoreactivity for substance P, calcitonin gene-related peptide, secretoneurin, and nitric oxide synthase throughout pregnancy. Many of these are small unmyelinated capsaicin-sensitive C-fibers. Substance P- (NK1-) and calcitonin gene-related peptide receptors were apparent on uterine cervix vasculature from pregnant, parturient, and postpartum rats. NK1 receptor mRNA was maximal at 20 days of pregnancy. Plasma extravasation of i.v. administered Evans Blue or Monastral Blue was most pronounced at parturition (shortly after NK1 mRNA is maximal); this was similar to plasma extravasation evoked by i.v. administration of substance P or capsaicin-treatment. This study revealed new data about the nervous system of the rat uterine cervix and that these nerves and their transmitters could very well be part of a neurogenic inflammatory process involved in cervical ripening.

Animals↗
PubMed1998

The antisense strategy applied to the study of dopamine D3 receptor functions in rat forebrain.

1. The authors have investigated the effects of a dopamine D3 receptor antisense oligodeoxynucleotide (ODN), on neuropeptides (neurotensin and dynorphin) and transcription factor (c-fos) mRNA levels in rat forebrain. 2. Intracerebroventricular injections of ODNs were made into the lateral ventricle (5 and 10 micrograms/h, for 5 days). Effect of antisense administration on dopamine D2 and D3 receptor binding were measured by means of receptor autoradiography. Neuropeptides and c-fos mRNA levels were evaluated by in situ hybridization using specific complementary RNA probes. 3. Dopamine D3 receptor densities were dose-dependently reduced in the shell of nucleus accumbens of rats that received the D3 antisense ODN. Sense and missense controls remained without effect. No significant effect was observed on D2 receptor binding in any of the ODN groups studied, as measured with [3H]raclopride binding. Concomitant reductions of dynorphin and neurotensin mRNA levels were observed in the shell of nucleus accumbens after D3 antisense ODN administration. Interestingly, the D3 antisense administration also reduced c-fos mRNA levels in the cingulate cortex of these animals. 4. The results show that D3 receptors may tonically regulate basal transcription factor, as well as neuropeptides, gene expression in the rat forebrain. These results clearly demonstrate that an antisense strategy could be useful to identify molecular targets under control of specific dopamine receptor subtypes.

Animals↗
PubMed1997

Quantitative reverse transcription and polymerase chain reaction analysis of oxytocin and vasopressin receptor mRNAs in the rat uterus near parturition.

Oxytocin receptors (OT-R) are known to be involved in the course of labor since a massive increase in OT-binding sites is observed in the uterus just before parturition. Vasopressin (AVP)-binding sites have also been observed and have been shown to mediate uterotonic responses. To determine exactly which subtypes of OT/AVP receptors are expressed in the rat uterus near parturition, we carried out absolute quantitations of the neurohypophysial hormone receptor (OT-R and the vasopressin receptors V1a-R, V1b-R and V2-R) mRNAs with an assay based on reverse transcription-polymerase chain reaction (RT-PCR) using in vitro transcribed mutated cRNAs as internal standards. The number of mRNA molecules/ng of total RNA was 35 +/- 6, 220 +/- 33 and 39 +/- 9 for OT-R (P < 0.01) and 16 +/- 1, 25 +/- 8 and 31 +/- 5 for V1a-R (P > 0.05) on day (D) 21, 22 and 23 of gestation (post-parturient), respectively. We did not detect V1b-R and V2-R mRNAs in the pregnant uterus. Therefore, the heterogeneity of OT and AVP receptors in the rat uterus can only be assigned to the presence of OT-R and V1a-R neurohypophysial hormone receptor subtypes, whereas V1b-R and V2-R can not be invoked. Only OT-R mRNA levels change in the uterus near parturition.

Animals↗
PubMed1997

A sensitive reverse transcriptase polymerase chain reaction assay for measuring the effects of dehydration and gestation on rat amounts of vasopressin and ocytocin mRNAs.

This study describes a competitive reverse transcriptase polymerase chain reaction (RT-PCR) method for assaying the amounts of vasopressin (AVP) and ocytocin (OT) mRNAs in the rat hypothalamus and uterus. Despite the low concentrations of these mRNAs, the RT-PCR method readily measured both AVP and OT mRNAs in the same sample. A common internal standard for both reactions was designed to quantify the reaction. Both AVP and OT mRNAs were readily quantified in a 75 ng sample of total RNA from the hypothalamus. Water deprivation stimulated AVP mRNA production 3-fold and OT mRNA production 1.7-fold in the hypothalamus. Gestation only influenced the amount of OT mRNA in the hypothalamus (3-fold increase) and uterus (38-fold increase). The amount of AVP mRNA in the hypothalamus remained unchanged and no AVP mRNA was detected in the uteri of either non-pregnant or pregnant rats. This competitive RT-PCR is a powerful tool that provides rapid and precise assays of AVP and OT mRNAs.

Animals↗
PubMed1998

mRNA coding for voltage-gated sodium channel beta2 subunit in rat central nervous system: cellular distribution and changes following kainate-induced seizures.

The cellular distribution of sodium channel beta2 subunit mRNA was examined in the central nervous system from adult Wistar rats using a non-radioactive in situ hybridization method with digoxigenin-labeled cRNA probes. The expression of the subunit was strong in cerebral and cerebellar cortex, in medulla oblongata and in the spinal cord whereas heterogeneous in hippocampus. The distribution was evaluated in hippocampus and cerebral cortex from 1 to 72 h after kainate injection and compared to control rats using densitometric analysis. In these areas, a transient increase was seen 1 h after the drug administration, followed, in the hippocampus, by a significant decrease. These variations differ from those we previously reported for alpha subunits and might play a role in cellular excitability changes occurring in the course of seizures.

Animals↗
PubMed1997

Continuous or pulsatile chronic D2 dopamine receptor agonist (U91356A) treatment of drug-naive 4-phenyl-1,2,3,6-tetrahydropyridine monkeys differentially regulates brain D1 and D2 receptor expression: in situ hybridization histochemical analysis.

The effect of a chronic D2 dopamine receptor agonist (U91356A) treatment on dopamine receptor gene expression in the brain of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned monkeys was investigated using quantitative in situ hybridization histochemistry. U91356A was administered to MPTP-monkeys for 27 days in a pulsatile (n=3) or continuous (n=3) schedule. Animals treated in a pulsatile mode showed progressive sensitization and developed dyskinesia; whereas with the continuous mode behavioural tolerance was observed but no dyskinesia developed. Untreated MPTP as well as naive control animals were also studied. The efficacy and uniformity of the MPTP effect was assessed by measures of dopamine concentrations by high performance liquid chromatography with electrochemical detection in the relevant brain areas. D1 and D2 receptor messenger RNAs levels were examined by in situ hybridization histochemistry using human complementary RNA probes. Intense specific labelling for D1 and D2 receptor messenger RNAs was measured in the caudate and putamen with a rostrocaudal gradient for D2 receptors and a lower density in the cortex for D1 receptors messenger RNA. D1 receptor mRNA levels in rostral striatum and cortex decreased whereas D2 receptor messenger RNA in caudal striatum increased in MPTP-monkeys compared to control animals. Continuous administration of U91356A reversed the MPTP-induced increase of D2 receptor messenger RNA, whereas the pulsatile administration did not significantly correct these messenger RNA changes. U91356A treatment whether continuous or pulsatile partially corrected the D1 receptor messenger RNA lesion-induced decrease in the striatum, whereas no correction was observed in the cortex. All MPTP-monkeys were extensively and similarly denervated suggesting that the D1 and D2 receptor expression changes following U91356A administration were treatment related. Our data show a lesion-induced imbalance of D1 (decrease) and D2 (increase) receptor messenger RNAs in the striatum of MPTP-monkeys. The response of these receptors to D1 agonist treatment showed receptor selectivity and was influenced by the time-course of drug delivery. Hence chronic continuous but not pulsatile administration of U91356A reversed the striatal D1 receptor messenger RNA increase.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1997

Detailed mapping of the histamine H2 receptor and its gene transcripts in guinea-pig brain.

Autoradiographic studies of the distribution of the histamine H2 receptor and its messenger RNAs were performed on serial frontal and a few sagittal sections of guinea-pig brain using [(125)I]iodoaminopotentidine for radioligand binding and a 33P-labelled complementary RNA probe for in situ hybridization, respectively. Both probes were validated by assessing non-specific labelling using non-radioactive competing H2 receptor ligands and a sense probe for binding sites and gene transcripts, respectively. In some areas, e.g., cerebral cortex, hippocampal complex or cerebellum, such studies were completed by identification of neurons expressing the H2 receptor messenger RNAs on emulsion-dipped sections. Nissl-stained sections from comparable levels were used to localize brain structures. In many brain areas, the distribution of the H2 receptor and its messenger RNAs appeared to parallel that known for histaminergic axons. For instance. high levels of both H2 receptor markers were detected in striatal and limbic areas known to receive abundant histaminergic projections. In contrast, in septum, hypothalamic, pontine and several thalamic nuclei, a comparatively low density of both H2 receptor markers was detected, suggesting that histamine actions in these areas are mediated by H1 and/or H3 receptors. Generally, the distribution of H2 receptor messenger RNA correlates well with that of [(125)I]iodoaminopotentidine binding sites, although some differences were observed. In a few regions (e.g., substantia nigra, locus coeruleus) high or moderate densities of binding sites were accompanied by a much more restricted expression of H2 receptor transcripts. Conversely, the mammillary region and the pontine nucleus exhibited higher levels of hybridization than of binding sites. In hippocampus, cerebral and cerebellar cortex there was a selective localization of the H2 receptor messenger RNA in the granule cells of dentate gyrus, pyramidal cells of the Ammon's horn and cerebral cortex, and Purkinje cells of cerebellum, whereas [(125)I]iodoaminopotentidine binding sites were located in layers where the dendritic trees of these messenger RNA-expressing neurons extend. The same discrepancy between messenger RNAs and binding sites suggests that striatonigral endings are endowed with the H2 receptor. The histamine H1 and H2 receptors both appear to be present in several brain areas, in some cases in a way suggesting their potential co-expression by the same neuronal populations, e.g., in granule and pyramidal cells in the hippocampal formation. This co-expression accounts for synergic responses, e.g., on cAMP generation, previously observed upon co-stimulation of both receptor subtypes. The widespread distribution of the H2 receptor, namely in thalamic nuclei or in telencephalic areas such as most layers of the cerebral cortex, together with its excitatory role previously established in electrophysiological studies, support its alleged function in mediating the histamine-driven control of arousal mechanisms. In addition, the detection of H2 receptor expression in brainstem areas from which other monoaminergic pathways involved in the control of states of sleep and wakefulness emanate, e.g., several raphe nuclei, locus coeruleus or substantia innominata, suggests possible interrelationships between all of these systems with highly divergent projections to the thalamus and telencephalon. The present mapping of the H2 receptor and its gene transcripts should facilitate neurochemical, neurophysiological and behavioural studies aimed at clarifying the role of histaminergic systems in brain.

Animals↗
PubMed1997

Molecular characterization and functional expression of a substance P receptor from the sympathetic ganglion of Rana catesbeiana.

Substance P is an important neuropeptide neurotransmitter in the central, autonomic and enteric nervous systems. In sympathetic ganglia, substance P is thought to play a role in modulating synaptic transmission. Release of substance P by neuronal stimulation or direct application of substance P to ganglionic neurons increases neuronal excitability. An amphibian substance P receptor complementary DNA has been cloned and characterized from bullfrog, Rana catesbeiana, sympathetic ganglion complementary DNA libraries. The deduced primary structure contains features indicative of a seven transmembrane domain G-protein-coupled receptor. The deduced protein sequence shows 69% identity to previously cloned mammalian substance P receptors. In situ hybridization analysis performed on bullfrog sympathetic ganglia using digoxigenin-labelled complementary RNA probe demonstrated that approximately 75% of the principal neurons displayed reaction product above background levels. Radioligand binding studies were performed on stably transfected cells with [(125)I]Tyr-1-substance P as the ligand. Substance P had an IC50 of 16 nM and the agonist potency profile was substance P>neurokinin A >> neurokinin B. The order of potency for three tachykinins to increase intracellular calcium when applied to a stably transfected clonal cell line was substance P>neurokinin A >> neurokinin B. This order of agonist potency also held for inhibition of the M-type potassium current in intact bullfrog sympathetic neurons. The non-peptide substance P antagonists CP-96345 and RP-67580 at concentrations that block mammalian substance P receptors had little or no effect on the responses to substance P at the bullfrog receptor. Overall, these results demonstrate that the cloned sequence has the features consistent with and characteristic of a substance P receptor. The results are discussed with reference to the established pharmacology of the bullfrog substance P receptor and known structure activity relationships of mammalian tachykinin receptors.

Animals↗
PubMed1999

Cellular and regional expression of glutamate dehydrogenase in the rat nervous system: non-radioactive in situ hybridization and comparative immunocytochemistry.

In the central nervous system glutamate dehydrogenase appears to be strongly involved in the metabolism of transmitter glutamate and plays a role in the pathogenesis of neurodegenerative disorders. In order to identify unequivocally the neural cell types expressing this enzyme, non-radioactive in situ hybridization, using a complementary RNA probe and oligonucleotide probes, was applied to sections of the rat central nervous system and, for comparison with peripheral neural cells, to cervical spinal ganglia. The results were complemented by immunocytochemical studies using a polyclonal antibody against purified glutamate dehydrodenase. Glutamate dehydrogenase messenger RNA was detectable at varying amounts in neurons and glial cells (i.e. astrocytes, oligodendrocytes, Bergmann glia, ependymal cells, epithelial cells of the plexus choroideus) throughout the central nervous system and in neurons and satellite cells of spinal ganglia. In some neuronal populations (e.g., pyramidal cells of the hippocampus, motoneurons of the spinal cord and spinal ganglia neurons) messenger RNA-labelling was higher than in other central nervous system neurons. This is remarkable because the immunostaining of neurons in the central nervous system regions studied was at best weak, whereas a predominantly high level of immunoreactivity was detected in astrocytes (and Bergmann glia). Thus, in neurons of the central nervous system, the detected levels of glutamate dehydrogenase messenger RNA and protein seem to be at variance whereas in peripheral neurons of spinal ganglia both in situ hybridization labelling and immunostaining are intense.

Animals↗
PubMed2000

Ontogeny of Rxt1, a vesicular "orphan" Na(+)/Cl(-)-dependent transporter, in the rat.

The developmental expression of the orphan Na(+)/Cl(-)-dependent transporter, Rxt1, was studied in the rat using a specific [(35)S]complementary RNA probe and affinity purified antibodies. Western blotting experiments allowed the detection of Rxt1 in brain as early as on embryonic day 16. After birth, the brain levels of Rxt1 increased dramatically up to a maximum around postnatal day 30 and then decreased slightly to the adult value. In situ hybridization experiments allowed the earliest detection of Rxt1 messenger RNA in the brain and spinal cord at embryonic day 14. In embryonic day 18 embryos, Rxt1 messenger RNA was present not only in the nervous system but also in the pituitary, the thymus and the heart. Immunoautoradiograms of whole embryo at embryonic days 16 and 18 showed high amounts of the Rxt1 protein in the spinal cord and brain. Moreover, at embryonic day 18, the orphan transporter was expressed in the thymus, heart and liver. At these ages, Rxt1 immunolabeling was localized in neurons of the subplate and in the ventricular zone of the brain. During early postnatal stages, Rxt1 messenger RNA expression demonstrated dynamic and complex changes until postnatal day 13. In particular, this transcript was relatively abundant in the striatum at postnatal days 3 and 5 and then decreased to very low levels after postnatal day 10. At the same period, Rxt1 immunostaining in the hippocampus and the cerebral cortex was observed all over the gray matter, in cell bodies as well as in the neuropil. Finally, the adult pattern was reached around postnatal day 13 for Rxt1 messenger RNA, but only at postnatal day 20 for the Rxt1 protein. The presence of Rxt1 messenger RNA and protein at embryonic stages and the high expression of the protein during synaptogenesis suggest that this vesicular "orphan" transporter is involved in the brain maturation process.

Aging↗
PubMed2004

Identification of genes involved in estrogenic action in the human prostate using microarray analysis.

Estrogens have profound effects on the developing prostate and are suspected to contribute to the development of benign prostatic hyperplasia, but the mechanism by which this hormone elicits its regulatory function still remains largely unknown. Using complementary RNA microarrays comprising approximately 10,000 oligonucleotide gene targets we compared differences in mRNA expression of estradiol-treated and untreated prostatic stromal cells in vitro. Based on a threshold of greater than twofold change, 228, 241, and 464 of the expressed genes were found to be regulated by estradiol after 10, 24, and 48 h of treatment, respectively. The secondary analysis of one estradiol-activated transcript, namely lipopolysaccharide-binding protein, and four estradiol-repressed genes, namely ras homolog gene family member E (RhoE/Rnd3), ubiquitin thiolesterase, interleukin 6, and interleukin 8 (IL-8), by real-time quantitative PCR confirmed the results of the microarray analysis. Moreover, IL-8 and RhoE were found to be down-regulated by estradiol at the protein level as well. We identified a set of genes involved in a wide range of cellular functions that are potentially important for understanding the molecular basis of estradiol action in the prostate.

Acute-Phase Proteins↗
PubMed2001

Expression domains in the skin of genes affected by the nude mutation and identified by gene expression profiling.

Although several genes affected by the nude mutation inactivating the transcription factor Whn have recently been identified, a comprehensive molecular analysis of the nude phenotype is still missing. Gene expression profiling of wildtype and nude mice back skin reveals several so far unknown differences in mRNA levels and demonstrates that microarray hybridization is ideal to identify even quantitative changes in expression. Some genes are upregulated in the absence of Whn. Most of the differentially expressed genes are downregulated in nude skin. Our results identify metallothionein IV among these genes. This is the first report on metallothionein IV expression in the murine hair follicle; its expression domain almost completely overlaps that of Whn.

Animals↗
PubMed2002

Expression pattern of dd4, a sole member of the d4 family of transcription factors in Drosophila melanogaster.

In vertebrates, three members of the d4 gene family code for proteins, which are believed to function as transcription factors and involved in regulation of various intracellular processes. One member of the family, ubi-d4/requiem is ubiquitously expressed gene and two other, neuro-d4 and cer-d4, are expressed predominantly in the neural tissues (Nucleic Acids Res. 20 (1992) 5579; Biochim. Biophys. Acta 14 (1992) 172; Mamm. Genome 11 (2000) 72; Mamm. Genome 12 (2001) 862). Typically, d4 proteins show distinct domain organisation with domain 2/3 in the N-terminal, Krüppel-type zinc finger in the central and two adjacent PHD-fingers (d4-domain) in the C-terminal part of the molecule. However, alternative splicing, which is responsible for complex expression patterns of both neurospecific members of the family, generates multiple protein isoforms lacking certain domains (Nucleic Acids Res. 20 (1992) 5579; Genomics 36 (1996) 174; Mamm. Genome 11 (2000) 72; Mamm. Genome 12 (2001) 862). Exact function of d4 proteins is unclear but their involvement in regulation of differentiation and apoptotic cell death has been proposed (J. Biol. Chem. 269 (1994) 29515; Mamm. Genome 11 (2000) 72; Mamm. Genome 12 (2001) 862). Here we identified a single gene, dd4, in the genome of Drosophila melanogaster, the protein product of which could be assigned to the d4 family. Expression of dd4 is regulated during Drosophila development, and is most prominent in syncytial embryos and later in the embryonic nervous and reproductive systems. In flies dd4 mRNA is found in most tissues but the highest level of expression is detected in ovaries.

Amino Acid Sequence↗