Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Although fetal growth is generally considered to be independent of pituitary growth hormone (GH), it is possible that pituitary GH plays a modulatory role in organ development or that a GH-like substance of non pituitary origin may influence fetal growth through the GH receptor. Accordingly, we have used immunohistochemistry, northern blot analysis, the reverse transcriptase-polymerase chain reaction and solution hybridization to study the ontogeny of the GH receptor/binding protein (BP) from the 12-day-old embryo (E12) to the E18 rat fetus. GH receptor/BP immunoreactivity was observed in all major organ systems of the E18 rat fetus and was not preferentially associated with any germ layer derivative. A general increase in GH receptor/BP immunoreactivity was evident from E12 to E18, with a marked increase occurring between E16 and E18. Hemangioblastic tissue was, however, strongly or intensely immunoreactive at all stages of development, as was the placenta. Most noteworthy of the other tissues expressing GH receptor/BP immunoreactivity by day 18 were skeletal and smooth muscle, chondroprogenitor cells, epithelial lining cells, neuronal ganglia, ependymal cells and the adrenal cortex. In the placenta, the most prominent immunoreactivity was associated with decidual cells. Total RNA was isolated from E12 to E18 rat fetuses and adult rat liver. Northern hybridization with a 35S-labelled rat GH receptor cRNA probe revealed that 3.9 kb and 1.2 kb transcripts complementary to the rat GH receptor riboprobe are present from at least E16. The existence of GH receptor mRNA at E12 and E14 was demonstrated by the polymerase chain reaction.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Prenatal expression of the growth hormone (GH) receptor/binding protein in the rat: a role for GH in embryonic and fetal development? 161 49

The precursor cells that form the enteric nervous system (ENS) are multipotent when they arrive in the gut from the neural crest. Their differentiation thus depends on signals from the enteric microenvironment. Crest-derived cells were isolated from the fetal rat bowel by immunoselection at E14 with NC-1/HNK-1 antibodies and secondary antibodies coupled to magnetic beads. NC-1/HNK-1-immunoreactive cells were enriched approximately 36-fold. The NC-1/HNK-1-selected population and the residual population were plated at equal cell density and maintained in a defined medium for 6-7 d. The total number of cells found in the cultures of the residual cells was three- to fourfold that in cultures of immunoselected cells. Neurotrophin-3 (NT-3), but not nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), or neurotrophin-4/5 (NT-4/5), was found to increase the proportion of neurons (neurofilament-immunoreactive or neuron-specific enolase-immunoreactive) or glia (S-100-immunoreactive) (from 6.6 +/- 0.9% to 15.2 +/- 1.4%; p < 0.001). This effect was concentration dependent (from 1 to 40 ng/ml) and observed only in the cultures of immunoselected cells. NT-3 also enhanced neurite outgrowth. NT-3 increased neither cell number nor bromodeoxyuridine incorporation and thus was not mitogenic. Exposure of immunoselected cells to NT-3 rapidly and transiently induced the appearance of nuclear Fos immunoreactivity. Transcripts coding for TrkC, the transducing receptor for NT-3, were identified in the fetal rat gut (E14-E16) and in the immunoselected population of cells using reverse transcriptase and the polymerase chain reaction. It is concluded that NT-3 specifically promotes the differentiation of enteric crest-derived cells as neurons or glia and may thus play a role in the development and/or maintenance of the ENS.
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PMID:Neurotrophin-3 induces neural crest-derived cells from fetal rat gut to develop in vitro as neurons or glia. 796 61

Quantitative receptor autoradiography with iodinated ligands, quantitative in situ hybridization histochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to describe the prenatal and early postnatal ontogeny (embryonic day 14 to postnatal day 7, or E14 to P7) of striatal D1 and D2 dopamine receptor binding sites and mRNA levels, respectively, in relation to the development of dopaminergic nigrostriatal innervation D1 dopamine receptor, measured by [125I]SCH23982 binding, and dopamine transporter binding sites, measured by [125I]RTI-55 binding, were present in low amounts beginning on E14 (2-3% and 0.3-0.6% of adult values, respectively) and increased slowly during the prenatal period. D2 receptor binding sites, measured with [125I]spiperone, were also detected on E14 but in higher relative quantities (17% of adult values) than D1 receptor and dopamine transporter binding sites at the same age. Other than abrupt declines in the late prenatal period for D1 and D2 receptor binding sites, all three binding sites increased throughout development and increased maximally between P7 and adulthood. On P5, both D1 and D2 receptors were functionally coupled to their respective G proteins, based on GTP-induced decreases in affinity of dopamine for [125I]SCH23982 and [125I]spiperone binding. D1 receptor mRNA was present in E14 striatal anlage, increased prenatally, declined on P0, then increased to a peak on P5, after which it declined to its lowest levels (20% of peak values) in the adult. In contrast, D2 receptor mRNA levels were presented also on E14, increased to a peak on P0, declined until P5, and increased thereafter to adulthood. Anatomically, nigrostriatal innervation and D1 and D2 receptor mRNA levels increased from the medial to lateral striatal quadrants. In contrast, D1 and D2 receptor binding site ontogency exhibited fairly homogenous distributions from E18 to P7. D1 and D2 receptor mRNAs appear to be expressed early in prenatal development before there is any significant dopaminergic innervation. In contrast, the majority of D1 and D2 receptor binding activity, representing expressed receptor proteins, develops in the postnatal period and correlates well with the increase in dopaminergic innervation. Intrinsic genetic programming is more likely to be responsible for D1 and D2 receptor gene transcription in striatal neuroblasts and newly born neurons, while factors derived from ingrowing dopaminergic afferents may direct post-transcriptional dopamine receptor development. The dissociation between the ontogeny of dopamine receptor binding sites and mRNAs suggests that the developmental regulation of D1 and D2 receptor synthesis is independent of D1 and D2 receptor gene transcription.
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PMID:Development of striatal dopaminergic function. I. Pre- and postnatal development of mRNAs and binding sites for striatal D1 (D1a) and D2 (D2a) receptors. 883 69

In the developing brain, histamine is one of the first neurotransmitters to appear. The concentration of histamine in the prenatal brain is fivefold that of adult levels. During the prenatal development a large transiently histamine-immunoreactive cell population distinct from the adult histaminergic system can be found within a subpopulation of the developing serotonergic raphe nuclei neurons. Also histamine-immunoreactive nerve fibers are widely distributed already during the prenatal development extending to the diencephalon, the thalamus, the cortex, and the spinal cord. Large numbers of histamine-containing mast cells also migrate into the brain during the late prenatal life. The wide distribution and high prenatal concentrations imply important functions for the histaminergic system during intrauterine development. However, little is known about the actual functions of histamine during development, and which of the histamine receptors are present in the prenatal rat brain is currently unknown. In the present study, we used in situ hybridization to study the distribution of H1-receptor (H1R) mRNA in the embryonic rat brain and spinal cord. H1R mRNA could be detected in rat brain and in spinal cord on embryonic day (E) 14, and the expression pattern seemed to partially localize in areas containing histamine-immunoreactive nerve fibers through E14-E20. H1R mRNA was also detected by reverse transcriptase polymerase chain reaction from embryonic brain samples and by Northern hybridization. The possible involvement of apoptosis in the disappearance of the developing transiently histaminergic system was studied by using apoptosis detection based on the terminal dUTP nick end labeling (TUNEL) technique and with c-Fos immunostaining. Although histamine immunoreactivity disappears dramatically from the developing raphe nuclei after E18, only occasional apoptotic nuclei could be seen in the histamine-immunoreactive cell bodies. The presence of H1R mRNA during the embryonic development renders it possible that histamine could exert an H1R-specific function at the time of the embryonic histamine peak.
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PMID:In situ detection of H1-receptor mRNA and absence of apoptosis in the transient histamine system of the embryonic rat brain. 955 Jan 46

Two nuclear receptors, Ad4BP/SF-1 and Dax-1, are essential regulators for development and function of the mammalian reproductive system. Similarity in expression sites, such as adrenal glands, gonads, pituitary, and hypothalamus, suggests a functional interaction, and the phenotype similarities were manifested in Ad4BP/SF-1-deficient mice and in cases of natural human mutations of Dax-1. In this study, quantitative reverse transcriptase polymerase chain reaction analyses revealed that expression profiles of Dax-1 in embryonic gonads are different between the two sexes and also from those of Ad4BP/SF-1. Immunohistochemical analyses clarified the spatial and temporal expressions of the Dax-1 protein during development of tissues composing the hypothalamic-pituitary-gonadal axis. During gonadal development, Dax-1 occurred after Ad4BP/SF-1 exhibiting a sexually dimorphic expression pattern at indifferent stages, indicating a possibility of Dax-1 involvement in earliest sex differentiation. When cord formation begins in the testis at embryonic day 12.5 (E12.5), Dax-1 was expressed strongly in Sertoli cells, but its expression level markedly decreased in Sertoli cells and increased in interstitial cells between E13.5 and E17.5. In the female, Dax-1 was strongly expressed in the entire ovarian primordium from E12.5 until E14.5, and then its expression level was decreased and limited to cells near the surface epithelium between E17.5 and postnatal day 0 (P0). During postnatal development of the testis, the variable staining of Dax-1 in Sertoli cells was detected as early as P7 and Dax-1-expressing Leydig cells became rare. In the postnatal ovary, Dax-1 expression was detected in granulosa cells with variable staining intensity, and occasionally in interstitial cells. During pituitary organogenesis, Dax-1 but not Ad4BP/SF-1 was expressed in the dorsal part of Rathke's pouch from E9.5. Later in development after E14.5, the distribution of Dax-1 overlapped with that of Ad4BP/SF-1, being restricted to gonadotropic cells in the anterior pituitary. In the ventromedial hypothalamus (VMH), Dax-1 and Ad4BP/SF-1 were mostly colocalized throughout the embryonic and postnatal development. Thus, the coexpression of Dax-1 and Ad4BP/SF-1 indicates their closely related functions in the development of the reproductive system. Furthermore, we noticed the presence of cells that express Dax-1 but not Ad4BP/SF-1, further indicating additional functions of Dax-1 in an Ad4BP/SF-1-independent molecular mechanism.
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PMID:Comparative localization of Dax-1 and Ad4BP/SF-1 during development of the hypothalamic-pituitary-gonadal axis suggests their closely related and distinct functions. 1130 69

In the pancreas, ligands of receptor tyrosine kinases (RTKs) are thought to be implicated in the development and function of the islets of Langerhans, which represent the endocrine part of the pancreas. In a previous study, we randomly screened by reverse transcriptase-polymerase chain reaction for RTKs expressed in the embryonic pancreas. One cDNA fragment that was cloned during this screen corresponded to the KIT receptor. The objective of the present study was to analyze the pattern of Kit expression in the pancreas. We demonstrated that Kit is expressed and functional in terms of signal transduction in the insulin-producing cell line INS-1. Indeed, upon treatment with the KIT ligand (KITL), the extracellular signal-regulated protein kinase was phosphorylated, and the expression of early responsive genes was induced. We also demonstrated that Kit mRNAs are present in fetal and adult rat islets. We next used mice that had integrated the lacZ reporter gene into the Kit locus. In these mice, beta-galactosidase (beta-gal) served as a convenient marker for expression of the endogenous Kit gene. Kit was found to be specifically transcribed in beta-cells (insulin-expressing cells), whereas no expression was found in other endocrine cell types or in the exocrine tissue. Interestingly, not all mature beta-cells expressed Kit, indicating that Kit is a marker of a subpopulation of beta-cells. Finally, by following beta-gal expression in the pancreas during fetal life, we found that at E14.5, Kit is expressed in both insulin- and glucagon-expressing cells present at that stage, and also in a specific cell population present in the epithelium that stained negative for endocrine markers. These data suggest that these Kit-positive/endocrine-negative cells could represent a subpopulation of endocrine cell precursors.
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PMID:Expression of the receptor tyrosine kinase KIT in mature beta-cells and in the pancreas in development. 1152 67

This report describes the expression and distribution pattern of RhoB GTPase in the developing mouse lens. RhoB expression was confirmed by sequencing an reverse transcriptase-polymerase chain reaction-generated DNA fragment of RhoB. Immunohistochemical analysis of RhoB revealed expression in the lens vesicle (both anterior and posterior vesicle) at embryonic day (E) 11.5, and in the epithelium and primary fibers of the E14.5 lens. Compared with the neonatal stage (day 1), where RhoB is detected in the entire lens (epithelium, primary, and secondary fibers), expression of this protein is restricted to the epithelial and outer cortical secondary fibers in postnatal lenses (from day 7 to day18). Interestingly, in E11.5 and E14.5 lenses, RhoB is localized predominantly in the lens, but not detectable in the retina, cornea, or other ocular tissues. RhoB expression appears to be down-regulated in the postnatal lens with concomitant up-regulation in the retina and cornea, compared with earlier stages of development (eyes of E11.5, E14.5, and neonatal mice). This study reveals the selective expression of RhoB in the lens during early eye development and suggests a potential role for this small GTPase in cytoskeletal reorganization associated with lens epithelial cell elongation and differentiation.
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PMID:Selective expression of the small GTPase RhoB in the early developing mouse lens. 1174 86

Temporal and spatial occurrence of hepatocyte growth factor (HGF) and its cognate receptor c-Met in the mouse mandibular development was investigated by immunohistochemistry and quantitative reverse transcriptase-polymerase chain reaction. HGF was first recognized in the mesenchymal cells of the first branchial arch at the 10th day of gestation (E10), before tongue formation, whereas HGF receptor (c-Met) -positive myogenic cells first appeared at E11 in the center of mandibles. By E12, HGF turned to be colocalized with c-Met in the differentiating tongue myoblasts. Between E14 and E16, HGF disappeared, whereas c-Met remained, in the tongue myoblasts. The levels of HGF mRNA in the developing tongue decreased in accordance with the increase of desmin mRNA levels from E11 to E17. These in vivo results strongly suggest that the HGF/c-Met system takes part in the earlier stages of tongue development. To elucidate this hypothesis, the antisense oligodeoxyribonucleotide (A-ODN) for mouse HGF mRNA was added to the organ culture system of mandible with serumless, defined medium. Mandibular arches from E10 mouse embryos were cultured at 37 degrees C for 10 days in the absence or presence of A-ODN, control (sense) oligonucleotide (C-ODN), or A-ODN plus recombinant HGF. In the control mandibular explants cultured without HGF or ODN, the anterior two-third of the tongue derived from the first branchial arch was formed. It contained abundant desmin-positive myoblasts and was equivalent to the tongue of E14-E15. In contrast, in the presence of A-ODN in the medium, neither the swelling nor myogenic cells were found in the tongue-forming region of explants, and myogenic cells accumulated behind the tongue-forming region. Such dysplasia of tongue was never induced in the presence of C-ODN or A-ODN plus recombinant HGF in the medium. The effect of A-ODN appeared to be developmental stage-specific, because tongue dysplasia occurred when A-ODN was present during the earlier 4 days but not during the later 4 days of the culture. Furthermore, recombinant HGF added to the culture without ODNs during the earlier 4 days caused elevation in the number of mitotic myoblasts. These results suggest that HGF regulates both the migration and proliferation of myogenic cells during the earlier stages of tongue development.
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PMID:Hepatocyte growth factor is essential for migration of myogenic cells and promotes their proliferation during the early periods of tongue morphogenesis in mouse embryos. 1183 82

Connexins (cx) constitute a family of transmembrane proteins that form gap junction channels allowing metabolic and electrical coupling of cellular networks. Initial studies on the expression of cx in the developing brain have suggested that cx may undergo dynamic changes and may possibly be implicated in synchronizing development and differentiation of neural progenitor cells and young neurons. We have investigated expression of cx26, cx32, cx43, and cx45 in the midbrain floor, where nigrostriatal dopaminergic neurons originate and differentiate. This neuron population is of major importance in regulating motor-functions. Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) revealed low levels of cx26-mRNA in the midbrain floor at E12, which gradually increased during pre- and postnatal development, reaching a maximum in the adult. Cx32-mRNA-levels reached a first peak at E16, and showed highest levels in adulthood. Cx43 was highly expressed at E12, decreased until E18, and subsequently increased again until adulthood. Cx45 mRNA was prominent at all developmental ages, but slightly decreased after the first postnatal week. Double-labeling for the dopaminergic neuronal marker tyrosine hydroxylase (TH), and cx-immunoreactivities (ir) evaluated by quantitative confocal laser microscopy revealed both distinct and similar developmental patterns for the individual cx investigated. Cx26 was highest at E14, decreased towards birth, and subsequently increased again reaching about 50% of the E14 level in the adult. Cx32-ir peaked at E16 and dropped to low levels after birth. Cx43-ir was highest at E12, decreased sharply at E14, reached its lowest levels at birth, but modestly increased again afterwards. Cx45-ir showed a biphasic pattern, with two prominent peaks at E12 and E18, followed by a massive postnatal decrease. Taken together, our results reveal that expression and ir of cx in the midbrain floor and dopaminergic neurons, respectively, follow cx-type specific patterns that temporally coincide with important steps of midbrain morphogenesis, as e.g. progenitor cell formation and migration (E12), early differentiation (E14-16), target encounter (E16-18) and postnatal functional maturation of the nigrostriatal system.
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PMID:Expression and developmental regulation of gap junction connexins cx26, cx32, cx43 and cx45 in the rat midbrain-floor. 1200 76

We investigated the involvement of hepatocyte growth factor (HGF) in salivary gland (SG) branching morphogenesis. The mouse submandibular gland (SMG) starts to develop at embryonic day 11.5-12 (E11.5-E12), and branching morphogenesis occurs in the area between the mandibular bone and tongue between E14 and E16.5. Real-time reverse transcriptase-polymerase chain reaction showed that the expression of the c-met/HGF receptor gene in SMG increased and peaked between E14 and E16.5, concomitant with epithelial branching, and high levels of HGF mRNA were detected in the surrounding mesenchyme at E14-E15.5. Although strong expression of the HGF and c-met transcripts was observed in the tongue muscles, this expression was limited at E13.5-E14.5. Serum-free organ cultures were established, in which SG rudiments that contained SMG and sublingual gland (SLG) primordia (explant 1) and SMG/SLG rudiments with peripheral tissue that included part of the tongue muscle (explant 2) were isolated from E13.5 or E14 embryos. Mesenchyme-free SMG epithelium was obtained by the removal of mesenchymal tissue from explant 1. In the explant 1 and 2 organ cultures, SMG/SLG rudiments showed growth and branching morphogenesis, while mesenchyme-free epithelium failed to grow. When E13.5 or E14 mesenchyme-free epithelium and a recombinant human HGF (rh-HGF) -soaked bead were placed on Matrigel, the epithelium migrated toward the bead and formed branches, while the E13 epithelium failed to branch. The exogenous application of rh-HGF and anti-HGF antibody to the SMG/SLG rudiment cultures resulted in stimulation and inhibition, respectively, of branching morphogenesis. However, the response of E13.5 SMG to rh-HGF was very weak, while the branching of E14 SMG was enhanced strongly by rh-HGF. The branching morphogenesis of SMG was also inhibited by the addition of either antisense HGF or c-met oligodeoxynucleotides to the cultures. The development of SMG in explant 2, which was significantly better than in explant 1, was comparable to that seen in vivo. Moreover, the expression of both HGF and c-Met in the SMG of explant 2 was higher than in the SMG of explant 1. These findings provide the first demonstration that the branching morphogenesis of SMG is regulated by interactions with the surrounding mesenchyme-derived HGF and c-met expression in SMG, which occur concomitant with epithelial branching. The present data also suggest that the HGF that is released transiently from tongue muscles may contribute to the rapid development of SMG at the branching stage.
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PMID:Involvement of hepatocyte growth factor in branching morphogenesis of murine salivary gland. 1451 89


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