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Discovery of semaphorin receptors, neuropilin and plexin, and their functions in neural development.

The semaphorin receptors neuropilin and plexin were initially identified as antigens for monoclonal antibodies MAb-A5 and MAb-B2, which bind to specific neuropiles and plexiform layers within the Xenopus tadpole nervous systems, several years before the discovery of the first semaphorin. This article provides an overview of how neuropilin and plexin were discovered. In addition, it describes the functions of neuropilin in the signaling of chemorepulsive activities of class 3 semaphorins and roles of neuropilin-mediated semaphorin activities in the directional guidance of the peripheral nervous system (PNS) and integration of the peripheral ganglia.

Animals↗

A hormone-sensitive communication system in an electric fish.

The electric communication system includes both special muscle-derived cells or electrocytes that produce species-typical electric signals, or electric organ discharges (EODs), and specialized sensory receptors, or electroreceptors, that encode the electric fields set up by EODs. Steroid hormones can influence the characteristic properties of both EODs and electroreceptors. Steroids appear to directly effect the anatomy and physiology of the electrocytes that generate an EOD. In contrast, the steroid effect on electroreceptors may be predominantly via an indirect mechanism whereby changes in the spectral characteristics of the EOD appear to induce changes in the spectral sensitivity of electroreceptors. Continued studies of electrosensory and electromotor systems will offer insights into the cellular bases for the development and evolution of steroid-sensitive pathways in the vertebrate nervous system.

Animals↗

Interactions of estrogen with the neurotrophins and their receptors during neural development.

We are interested in examining mechanisms underlying estrogen actions during neuronal differentiation in the central nervous system (CNS). Our research has focused on one possible mechanism, the developmental interactions between estrogen and the neurotrophins (nerve growth factor [NGF], brain derived neurotrophic factor [BDNF] and neurotrophin-3 [NT-3]). Using combined isotopic and non-isotopic in situ hybridization, we found that neurons in developmental estrogen targets (e.g., the cerebral cortex), co-localized mRNAs for the neurotrophins (NGF or BDNF) with their cognate receptors (p75NGFR [the pan-neurotrophin receptor] and trkA or trkB [the tyrosine kinase receptors]), suggesting a localization of neurotrophin-autocrine loops to these estrogen-sensitive neurons. In contrast, the basal forebrain, which is estrogen-sensitive in the adult and during development, only expressed neurotrophin receptor mRNAs, suggesting that this region was not an autocrine neurotrophin target. We examined the potential for developmental estrogen-neurotrophin interactions, using a model neurotrophin-sensitive system, i.e., differentiating PC12 cells. NGF significantly increased estrogen receptor density in PC12 cells. Reciprocally, estrogen up-regulated trkA mRNA and transiently down-regulated p75NGFR mRNA, suggesting that estrogen may increase the efficiency of NGF binding in PC12 cells. Similar estrogen-dependent regulation of NGF receptor mRNAs were also observed in the adult dorsal root ganglia, suggesting that estrogen may regulate NGF sensitivity in adult neurotrophin targets as well. Such estrogen-neurotrophin interactions may have an important role during differentiation and in the adult, following injury.

Animals↗

POU domain factors in neural development.

Transcription factors serve critical roles in the progressive development of general body plan, organ commitment, and finally, specific cell types. Comparison of the biological roles of a series of individual members within a family permits some generalizations to be made regarding the developmental events that are likely to be regulated by a particular class of transcription factors. Here, we evidence that the developmental functions of the family of transcription factors characterized by the POU DNA binding motif exerts roles in mammalian development. The POU domain family of transcription factors was defined following the observation that the products of three mammalian genes, Pit-1, Oct-1, and Oct-2, and the protein encoded by the C. elegans gene unc-86, shared a region of homology, known as the POU domain. The POU domain is a bipartite DNA binding domain, consisting of two highly conserved regions, tethered by a variable linker. The approximately 75 amino acid N-terminal region was called the POU-specific domain and the C-terminal 60 amino acid region, the POU-homeodomain. High-affinity site-specific DNA binding by POU domain transcription factors requires both the POU-specific and the POU-homeodomain. Resolution of the crystal structures of Oct-1 and Pit-1 POU domains bound to DNA as a monomer and homodimer, respectively, confirmed several of the in vitro findings regarding interactions of this bipartite DNA binding domain with DNA and has provided important information regarding the flexibility and versatility of POU domain proteins. Overall the crystal structure of a monomer of the Oct-1 POU domain bound to the octamer element was similar to that predicted by the NMR solution structures of the POU-specific domain and the POU-homeodomain in isolation, with the POU-specific domain consists of four alpha helices, with the second and third helices forming a structure similar to the helix-turn-helix motif of the lambda and 434 repressors; several of the DNA base contacts are also conserved. A homodimer of the Pit-1 POU domain was crystallized bound to a Pit-1 dimer DNA element that is closely related to a site in the proximal promoter of the prolactin gene. The structure of the Pit-1 POU domain on DNA is very similar to that of Oct-1, and the Pit-1 POU-homeodomain/DNA structure is strikingly similar to that of other homeodomains, including the Oct-1 POU-homeodomain. The DNA contacts made by the Pit-1 POU-specific domain are also similar to those of Oct-1 and conserved with many made by the prokaryotic repressors. In the Oct-1 crystal, the POU-specific domain recognizes a GCAT half-site, while the corresponding sequence recognized by the Pit-1 POU-specific domain, GTAT, is on the opposing strand. As a result, the orientation of the Pit-1 POU-specific domain relative to the POU-homeodomain is flipped, as compared to the Oct-1 crystal structure, indicating the remarkable flexibility of the POU-specific domain in adapting to variations in sequence within the site. Also in contrast to the Oct-1 monomer structure is the observation that the POU-specific and POU-homeodomain of each Pit-1 molecule make major groove contacts on the same face of the DNA, consistent with the constraints imposed by its 15 amino acid linker. As a result, the Pit-1 POU domain homodimer essentially surrounds its DNA binding site. In the Pit-1 POU domain homodimer the dimerization interface is formed between the C-terminal end of helix 3 of the POU-homeodomain of one Pit-1 molecule and the N-terminus of helix 1 and the loop between helices 3 and 4 of the POU-specific domain of the other Pit-1 molecule. In contrast to other homeodomain crystal structures, the C-terminus of helix 3 in the Pit-1 POU-homeo-domain has an extended structure. (ABSTRACT TRUNCATED)

Animals↗

Transient serotonin-immunoreactive neurons coincide with a critical period of neural development in coho salmon (Oncorhynchus kisutch).

In coho salmon (Oncorhynchus kisutch), smolt transformation has been shown to be associated with sequential surges of neurotransmitters in the brain. In order to determine if the surge of serotonin (5-HT) is correlated with structural changes, we have used immunocytochemistry to observe changes in the serotonin immunoreactivity before, during and after the 5-HT surge. The following stages were studied: 12-month-old freshwater presmolts, 17-month-old freshwater presmolts, 18-month-old saltwater smolts, 19-month-old saltwater postsmolt, 24-month-old postsmolt, and adult spawners. In the 17-month-old samples, but not at any other stage, we found a set of transient (serotonin-immunoreactive) 5-HT-immunoreactive neurons in the lateral preoptic area, as well as a discrete population of 5-HT-immunoreactive neurons in the lateral part of the dorsal right habenular nucleus. In addition, a higher density of serotonergic fibers was found in the telencephalon at this stage compared to the following two stages. Since the transient 5-HT-immunoreactive structures presented here do not appear simultaneously with the 5-HT total brain concentration surge, we conclude that they are unlikely to be the source of the 5-HT surge, but are probably related to other developmental changes in the brain associated with smolt transformation.

Animals↗

Mutational analysis of the Caenorhabditis elegans ankyrin gene unc-44 demonstrates that the large spliceoform is critical for neural development.

In Caenorhabditis elegans, unc-44 mutations affect axonal outgrowth and guidance, leading to locomotory defects. The wild-type unc-44 gene encodes a family of ankyrin proteins, which, in addition to the conventional ankyrins, includes a novel ankyrin isoform with an extended C-terminal domain, referred to AO13 ankyrin. Six spontaneous unc-44 mutations and their reversions were analyzed in order to localize regions critical for gene function. The q331::Tc1 and rh1013::Tc1 mutations were mapped to the portion of the gene encoding the conventional ankyrins, mn339 had an uncharacterized 2-kb insertion in the serine/threonine/glutamic acid/proline-rich (STEP) repeat block 5, st200::Tc5(variant) and rh1042::Tc1 were localized near the C-terminus, and mn259 resulted from two Tc1 insertions, one in STEP block 6 and the other near the C-terminus. Tc1 excisions in several revertants resulted either in the restoration of the wild-type sequence, or were associated with small in-frame deletions or insertions. Reversion of mn339 resulted in the net excision of 2463 bp of genomic DNA, including the region encoding parts of STEP blocks 5 and 6 and the intervening hydrophobic region. Interestingly, additional Tc1 insertions at a 5' exon/intron boundary were found in revertants of st200 and rh1042. Reversion of the st200::Tc5 mutation resulted in excision of the Tc5 element, and the insertion of two copies of Tc1 at different sites. The wild-type unc-44 gene produces multiple transcripts - shorter RNAs determined to be approximately 1, 3.2, 5, 6, and 7 kb long, and two large transcripts estimated to be 22 and 26 kb in length. The largest transcripts were affected by all unc-44 mutations and are proposed to be essential for axonal outgrowth and guidance.

Alternative Splicing↗

Neural development in children: a neurophysiological study.

In adult human subjects torque motor imposed angular displacements of the upper limb joints result in two major reflex EMG components as identified by their latency. The shorter latency (SL) component is probably the spinal stretch reflex and the longer latency (LL) reflex component may involve supraspinal structures. Tendon jerk, arising via the spinal reflex pathway, is present in children at birth. In this study the presence and the development of the LL component in wrist flexors was investigated in children between the ages of 2 and 13 years. Kinaesthetic reaction times were tested simultaneously. In young children, the duration of the LL component was much longer than that in adults. The duration decreased slowly from 2 to 6 years, following which a relatively abrupt decrease between the ages of 6 and 8 years took place. After 8 years of age responses looked more adult-like. Kinaesthetic reaction times attained adult-like values after 10 years of age. Various possibilities underlying these observations are discussed.

Adolescent↗

Beta adrenergic modulation of cardiac rhythm in a rat model of altered sympathetic neural development.

We previously have shown that treatment of neonatal rats (days 1-10) with Nerve Growth Factor (NGF) or its antibody (Ab) modifies alpha-adrenergic receptor-effector coupling, such that innervated hearts at day 10 show high levels of a 41 kDa GTP regulatory protein (G protein) that is a substrate for pertussis toxin and that links the alpha 1-receptor to the Na/K pump. This receptor-effector pathway results in alpha adrenergic-induced decreases in automaticity. In contrast, non-innervated hearts at day 10 show lower levels of the pertussis toxin sensitive G-protein and increases in automaticity induced by alpha-agonist. We now report the effects of administration of NGF, Ab or placebo on beta-adrenergic receptor-effector coupling in neonatal rats. Rats were administered NGF, Ab or placebo on days 1-10 of life. On day 10, the beta-receptor number and affinity and the stimulatory G-protein, Gs, were equivalent across groups. Moreover, the ventricular automatic response to beta-adrenergic receptor stimulation was equivalent across groups suggesting there was no change in receptor-effector coupling as a result of the difference in innervation. These results on beta-adrenergic receptor-effector coupling considered in light of our prior studies on alpha-adrenergic coupling suggest that the development of sympathetic innervation is more a determinant of alpha than beta adrenergic modulation of ventricular rhythm.

Adrenergic beta-Agonists↗