Embryo emergent: elucidating the cell biology of development. The Santa Cruz Conference on Developmental Biology 2000.
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Animals↗
Biomedical subjects
M Halpern
Publications and source records attributed to M Halpern.
Organization of the ophidian amygdala: chemosensory pathways to the hypothalamus.
Although recent studies in squamate reptiles have importantly clarified how chemical information is processed in the reptilian brain, how the amygdala relays chemosensory inputs to the hypothalamus to influence chemically guided behaviors is still poorly documented. To identify these chemosensory pathways, the amygdalo-hypothalamic projections, intra-amygdaloid circuitry and afferents from the lateral cortex (LC) to the amygdala were investigated by injecting conjugated dextran-amines into the hypothalamus, amygdala, and LC of garter snakes. The amygdala was divided into olfactory recipient (ventral anterior and external amygdalae), vomeronasal recipient (nucleus sphericus, NS, and medial amygdala, MA), and nonchemosensory (e.g., posterior dorsal ventricular ridge, PDVR, and dorsolateral amygdaloid nucleus, DLA) subdivisions. Rostroventral (LCrv) and dorsocaudal subdivisions of the LC were distinguished. In addition to receiving afferents from the main olfactory bulb, the olfactory amygdala receives afferents from NS and projects to the NS, PDVR, and dorsal hypothalamus. The NS has only a minor projection to the lateral hypothalamus, whereas the MA, which receives afferents from the LCrv and NS, has projections to the ventromedial hypothalamic (VMH) and lateral posterior hypothalamic nuclei. Among the nonchemosensory amygdaloid structures, the PDVR receives afferents from the LCrv and the olfactory amygdala and projects to the VMH, whereas DLA receives afferents from the LCrv and NS, and projects to the periventricular hypothalamus. These results substantially clarify the olfactory and vomeronasal tertiary connections and demonstrate that parts of the nonchemosensory amygdala play a major role in relaying chemosensory information to the hypothalamus.
Amygdala↗
Differential centrifugal afferents to the anterior and posterior accessory olfactory bulb.
The centrifugal afferents to the anterior and posterior divisions of the accessory olfactory bulb (AOB) were investigated after iontophoretic injections of dextranamines. Injections affecting the anterior and posterior or just the posterior division of the AOB gave rise to retrogradely labeled cells in the bed nuclei of the accessory olfactory tract and stria terminalis and in the anterodorsal medial and posteromedial cortical (PMCo) amygdaloid nuclei. Injections restricted to the anterior division of the AOB yielded similar results, although no cells were observed in the PMCo. These results demonstrate differential centrifugal inputs to the anterior and posterior divisions of the AOB, probably to the granular layer, and provide further support for the hypothesis of a functionally segregated vomeronasal system.
Afferent Pathways↗
Signal transduction in the vomeronasal organ of garter snakes: ligand-receptor binding-mediated protein phosphorylation.
The vomeronasal (VN) system of garter snakes plays an important role in several species-typical behaviors, such as prey recognition and responding to courtship pheromones. We (X.C. Jiang et al., J. Biol. Chem. 265 (1990) 8736-8744 and Y. Luo et al., J. Biol. Chem. 269 (1994) 16867-16877) have demonstrated previously that in the snake VN sensory epithelium, the chemoattractant ES20, a 20-kDa glycoprotein derived from electric shock-induced earthworm secretion, binds to its receptor which is coupled to PTX-sensitive G-proteins. Such binding results in elevated levels of IP3. We now report that ES20-receptor binding regulates the phosphorylation of two membrane-bound proteins with molecular masses of 42- and 44-kDa (p42/44) in both intact and cell-free preparations of the VN sensory epithelium. ES20 and DAG regulate the phosphorylation of p42/44 in a similar manner. ES20-receptor binding-mediated phosphorylation of p42/44 is rapid and transient, reaching a peak value within 40 seconds and decaying thereafter. Phosphorylation of p42/44 appears to be regulated by the countervailing actions of a specific membrane-bound protein kinase and a protein phosphatase. The phosphorylation of these membrane-bound proteins significantly reduces the activity of G-proteins as evidenced by a decrease in GTPase activity, but has little effect on ligand-receptor binding. These findings suggest that p42/44 play a role in modulating the signal transduction induced by ES20 in the vomeronasal system.
Animals↗
Expression of the apoptosis-related oncogenes bcl-2, bax, and p53 in Merkel cell carcinoma: can they predict treatment response and clinical outcome?
Chemotherapy and radiation therapy act predominantly through the induction of apoptosis in malignancies. Merkel cell carcinoma, an aggressive malignancy with prominent apoptosis, has proved to be sensitive to both modes to a certain degree. We used immunohistochemical methods to examine 25 Merkel cell carcinomas and 8 of their lymph node metastases to assess the status of the antiapoptotic gene bcl-2 and 2 proapoptotic genes, wild-type p53 and bax. All tumors showed prominent bax immunopositivity; 76% were positive for bcl-2, and only 28% were positive for p53, the latter presumably reflecting mutated p53. No statistically significant relationship was found between tumor immunopositivity and therapy response or survival. The widespread bax immunopositivity and the apparently low rate of p53 mutations, as suggested by the low rate of p53 immunopositivity, may be related to the presence of prominent apoptosis in Merkel cell carcinoma. The finding of bcl-2 immunopositivity in 76% of the tumors suggests that some of the tumor cells may be resistant to apoptosis-inducing agents.
Adult↗
Differential projections from the anterior and posterior divisions of the accessory olfactory bulb to the medial amygdala in the opossum, Monodelphis domestica.
The vomeronasal sensory epithelium of mammals contains apical and basal cell populations expressing different G proteins and putative pheromone receptors, which project, respectively, to the anterior and posterior divisions of the accessory olfactory bulb (AOB). In order to analyse whether these segregated pathways are preserved in the connections between the AOB and the amygdala, conjugated dextran-amines were iontophoretically injected into the anterior and posterior divisions of the AOB. We found that efferent projections from both divisions essentially overlap throughout the vomeronasal recipient amygdala. In the medial amygdaloid complex, both divisions project to lamina 1A of layer 1 of the anterodorsal, anteroventral, posterodorsal and posteroventral nuclei. The posterior division alone, however, projects to lamina 1B and layers 2 and 3 of the anterodorsal, anteroventral and posteroventral nuclei. These results constitute a link between molecular, anatomical and functional approaches on the study of the vomeronasal system. Molecular and functional studies support that the two segregated pathways between the vomeronasal organ and the AOB are functionally different. Similarly, the anatomical approaches to the further connections of this system indicate that the medial amygdala possesses ventral and dorsal divisions that are hodologically and functionally different. The present results demonstrate a differential projection from the posterior AOB to the ventral division of the medial amygdala. These findings indicate that the segregated pathways of the vomeronasal system continue to the level of the amygdala, and they provide some clues about the functional implications.
Amygdala↗
Chloramine and copper sulfate as control agents of planktonic larvae of Chironomus luridus in water supply systems.
The latest approach to control of midge larvae in drinking-water supplies is suppression of the planktonic 1st-stage larvae, by using 2 disinfectants, chloramine and copper sulfate. The median lethal concentration for 24-h exposure of the 1st-stage larvae of Chironomus luridus to chloramine and copper sulfate individually was 0.51 and 0.38 mg/liter, respectively. The increase of copper sulfate to 0.5 mg of copper per liter to water containing chloramine (0.5 mg/liter) created a synergistic reaction that resulted in 96% (+/-8% SD) mortality of the planktonic larvae. This treatment may serve as an effective control of 1st-stage larvae in municipal drinking-water supplies.
Animals↗
Development of NADPH-diaphorase expression in chemosensory systems of the opossum, Monodelphis domestica.
Using NADPH-diaphorase histochemistry, the present study describes development of olfactory and vomeronasal systems in postnatal opossums, Monodelphis domestica. NADPH-dependent staining is absent at and around the time of birth. By 2 weeks of age and through adulthood, intense staining is seen along the luminal surface of the olfactory epithelium (OE) and of the vomeronasal sensory epithelium (VNE), as well as in Bowman's glands of the OE. Staining of the adult VNE is not homogeneous; it is restricted to the superficial 2/3 of the epithelium. At 2 weeks of age, staining in the brain is seen only at the surface of the ventricles and in blood vessels. At 1 month of age and through adulthood, staining of varying intensity is seen in individual olfactory bulb glomeruli, although the incoming olfactory axons are relatively unstained. Interestingly, whereas at 30 days of age, staining of the accessory olfactory bulb (AOB) glomeruli is uniform, 2 weeks later and in the adult, NADPH staining is concentrated in the rostral half, with little or no staining observed in the posterior portion. Darkly stained periglomerular cells are seen throughout the extent of the differentially-stained glomerular layer. From 30 days of age and through adulthood, intense NADPH staining is also observed in the islands of Calleja, as well as in cells of the dorsal cortex, often associated with the path of the rostral migratory stream.
Aging↗
Neurogenesis and migration of receptor neurons in the vomeronasal sensory epithelium in the opossum, Monodelphis domestica.
The sensory epithelium of the vomeronasal organ (VNO) contains primary chemosensory receptor neurons that project to the accessory olfactory bulb (AOB). In the present study, neurogenesis and cell migration in the sensory epithelium of the VNO were analyzed in opossums (Monodelphis domestica) by using bromodeoxyuridine (BrdU) labeling. 1) In the VNO of normal adult opossums, BrdU labeled a small number of cells localized in the basal region of the sensory epithelium. After 1 or 2 weeks of survival, the labeled cells appeared in the receptor cell layers and became receptor neurons, as indicated by coexpression of the G proteins G(i alpha2) or G(o alpha). 2) In the VNO in which the receptor neurons had been destroyed by removing the AOB, the number of BrdU-labeled cells in the reconstituting sensory epithelium was greatly increased compared with that in the intact VNO. The labeled cells were also located in the basal region of the sensory epithelium. 3) In the developing VNO (at postnatal day 10), more cells in the basal region of the sensory epithelium were labeled than in the adult VNO, indicating rapid cell proliferation; and there appeared to be more labeled cells in the basal region near the margins of the sensory epithelium where it meets the nonsensory epithelium. These observations demonstrate that, in the opossum VNO, there is a population of proliferating cells in the basal region close to the basal lamina in the sensory epithelium. The newly generated neurons in the basal region migrate vertically into the receptor cell layer.
Animals↗
Chemosignal transduction in the vomeronasal organ of garter snakes: cloning of a gene encoding adenylate cyclase from the vomeronasal organ of garter snakes.
We previously reported that ES20-receptor binding activates phosphoinositide (PI) turnover, resulting in an increase in inositol-1,4,5-trisphosphate, which in turn mobilizes intracellularly stored calcium in the vomeronasal (VN) sensory epithelium of garter snakes. We also found that the activity of adenylate cyclase (AC) in the VN organ is very sensitive to Ca2+ but insensitive to calmodulin regulation. A 250-bp fragment of adenylate cyclase type VI (AC-VI) was obtained from brain cDNA of garter snake by RT-PCR with degenerate primers. The 250-bp fragments were amplified, cloned, and sequenced. Both Northern blot and RNase protection assays revealed that the vomeronasal organ (VNO) and brain contained more abundance of AC type VI than the main olfactory epithelium. A 3.8-kb cDNA was then cloned from the vomeronasal cDNA library of garter snakes and sequenced. The 5' cDNA was obtained by means of 5' RACE PCR and sequenced. We have successfully cloned a 5200-nucleotide cDNA from VNO of garter snakes containing an open reading frame++ encoding 1150 amino acids of AC-VI protein. The vomeronasal AC is termed AC(VN) . AC(VN) shows a high degree of homology with type VI AC of rat, mouse, or human. In situ hybridization with digoxigenin-labeled cRNA demonstrated that AC(VN) mRNA was abundant in the sensory epithelium but not in the nonsensory epithelium of the mushroom body of the vomeronasal organ of garter snakes.
Adenylyl Cyclases↗
Effects of bacterial DNA on cytokine production by (NZB/NZW)F1 mice.
Microbial DNA has multiple immune effects including the capacity to induce polyclonal B cell activation and cytokine production in normal mice. We recently described the accelerated induction of anti-DNA Abs in NZB/NZW mice immunized with Escherichia coli (EC) dsDNA; paradoxically these mice developed less renal disease than unimmunized mice or mice immunized with calf thymus DNA. We postulated that alterations in cytokine production induced by bacterial DNA may play a key role in renal protection. To determine the effect of bacterial DNA on cytokine production in NZB/NZW mice, we measured the serum cytokine levels, cell culture supernatant cytokine levels, and number of cytokine-producing splenocytes in NZB/NZW mice injected with EC DNA, calf thymus DNA, or an immune active oligonucleotide. There was a 10- to 25-fold increase in the number of cells secreting IFN-gamma compared with IL-4 in mice immunized with EC DNA. IL-12-secreting cells were also increased by bacterial DNA immunization. In parallel with the increase in IFN-gamma secreting cells, there was a significant rise in serum IFN-gamma levels in mice receiving EC DNA. These results indicate that EC DNA modulates systemic cytokine levels in NZB/NZW mice, selectively increasing IL-12 and IFN-gamma while decreasing IL-4 production. The cytokine response of NZB/NZW mice to bacterial DNA may be of significance in disease pathogenesis and relevant to the treatment of lupus-like disease.
Animals↗
Segregated pathways in the vomeronasal system.
The vomeronasal system of mammals is chemoarchitecturally dichotomous. Two populations of receptor cells have been identified in the vomeronasal sensory epithelium based on the family of receptor proteins they express on their membranes. These two receptor cell populations express different G-proteins: the more basal population expresses Goalpha and the more apical population expresses Gialpha2. The Goalpha-expressing receptor cells project their axons to the posterior accessory olfactory bulb (AOB) whereas the Gialpha2-expressing cells project their axons to the anterior AOB. In all mammals studied to date, the anterior AOB is Gialpha2-positive and the posterior AOB is Goalpha-positive. These two parts of the AOB are also chemoarchitecturally heterogeneous with respect to their carbohydrate content as revealed both with lectin binding and immunoreactivity to monoclonal antibodies raised against carbohydrate moieties. However, species differences have been observed with respect to lectin binding, as with NADPH-diaphorase reactions and OMP immunoreactivity. Recent studies indicate that there are physiological and behavioral correlates to the dichotomy within the vomeronasal system.
Animals↗
Analysis of ras gene mutational spectra in epidermal papillomas from K6/ODC transgenic mice.
In standard mouse strains, a high proportion (more than 90%) of epidermal tumors produced by initiation with 7,12-dimethylbenz[a]anthracene and promotion with a variety of chemical agents contain an activating mutation in codon 61 (A182-->T) of the c-Ha-ras gene. We analyzed the ras mutational spectra in 69 tumors induced by DMBA in a unique transgenic model, the K6/ODC mouse. In this model, low-dose DMBA treatment is sufficient per se for tumor induction, so tumor promotion with chemical agents is not required. In contrast to previous studies in standard mouse strains, our study showed that less than 50% of epidermal tumors from K6/ODC mice contained an activating codon 61 c-Ha-ras mutation (A182-->T). This result was obtained in mice initiated either as newborns (when the transgene is not expressed) or as adults (when the transgene is fully expressed). Analysis of other codon hot-spots and other ras genes revealed the presence of three codon 12 and 20 codon 61 (A182-->T) mutations in the c-Ki-ras gene in the 36 tumors that did not have c-Ha-ras mutations. We concluded that promotion in this model, by means of constitutive ornithine decarboxylase expression, causes the clonal expansion of a population of initiated cells not promoted by chemical agents.
9,10-Dimethyl-1,2-benzanthracene↗
Leiomyoma of the urinary bladder.
Leiomyoma of the urinary bladder is a rarity but should be considered in the differential diagnosis of intramural neoplasm. We report a case illustrating clinical and pathological features in particular the immunohistochemistry. Etiology and differential diagnosis are discussed.
Diagnosis, Differential↗
Heterogeneity in the accessory olfactory system.
The mammalian accessory olfactory bulb (AOB) is chemoarchitecturally heterogeneous in that it stains differentially with a number of markers; the receptor cells that project to the AOB are similarly heterogeneous. What is the significance of this heterogeneity? We have found that the AOB of the gray, short-tailed opossum, Monodelphis domestica, stains differentially with a number of 'markers': antibodies to olfactory marker protein (OMP) and the alpha subunit of the G protein Gi2, the lectin of Vicia villosa and NADPH-diaphorase. These markers stain the rostral AOB more strongly than the caudal AOB whereas, the G protein subunit G(o) alpha is located predominantly in the posterior subdivision of the AOB. This heterogeneity in the chemoarchitecture of the AOB may reflect a fundamental organizational dichotomy within the vomeronasal system that corresponds to a functional dichotomy. The vomeronasal sensory epithelium also exhibits a chemoarchitectural heterogeneity: receptor cells in the basal third are G(o) alpha-immunoreactive whereas the cells in the middle third are Gi2 alpha-immunoreactive. Tracing studies using WGA-HRP demonstrate that the neurons in the middle third of the vomeronasal sensory epithelium project their axons to the anterior AOB whereas those in the basal third appear to project to the posterior AOB.
Animals↗
The characteristics of the electrovomeronasogram: its loss following vomeronasal axotomy in the garter snake.
Electrovomeronasogram (EVG) recordings were made from adult garter snakes, Thamnophis sirtalis. Stimulation of vomeronasal epithelium with a stimulus prepared from prey, earthworm electric shock secretion (ESS), evoked EVG response in a dose-dependent manner. The magnitude of the EVG response to ESS was remarkably larger than n-amyl acetate and glutamate, which elicited insignificant responses, supporting the idea that the vomeronasal system is differentially sensitive to liquid delivery of biologically significant chemical stimuli. Fourteen days following vomeronasal axotomy, the magnitudes of the EVG responses of animals which received bilateral axotomy without cauterization or with cauterization was -0.19+/-0.07 mV or -0.05+/-0.02 mV respectively, compared with the normal EVG response of -0.41+/-0.10 mV. The epithelia of animals which received bilateral axotomy without cauterization exhibited remarkable degeneration of the bipolar neurons. Maximal depletion of bipolar neurons occurred in the epithelia denervated with cauterization, though the difference between cell densities in vomeronasal neuron layers in these epithelia was not statistically significant. The present results clearly indicate that the fewer neurons the epithelium contains, the smaller EVG response it generates, suggesting that the receptor neurons are the primary origin of EVG responses.
Animals↗
Shock chloramination: potential treatment for Chironomidae (Diptera) larvae nuisance abatement in water supply systems.
In the early 1990s, infestations of midge larvae (Chironomidae, Chironomus sp.) were discovered in the potable water system of Tel Aviv, Israel. Control measures, such as draining and cleaning tanks, spraying water into the tank's air space, and electrocution traps of midge adults, were either inadequate or ineffective. In this system, monochloramine concentrations of up to 0.75 mg/liter are used routinely as a secondary disinfectant. This chemical was tested in the laboratory as a toxicant of midge larvae. The mortality of 4th instar midge larvae after short exposure to high chloramine concentrations (LC50 values of 32 mg/liter for 75 min) suggested the efficacy of instituting a Shock Chloramination treatment program. Tanks were partially drained until they contained only 20 cm of water and were then temporarily disconnected. Chloramine was added to this water to produce a concentration of approximately 70 mg/liter for 1-2 h. Subsequently, all dead chironomids were flushed out, and the tank was refilled to attain the operational volume of water. A 2nd identical treatment of water in the tank was suggested 7 d later to kill midges from reproductive adults and egg-masses that survived the 1st treatment. This treatment program was tested in commercial covered tanks and gave complete control of these pests for 6-10 wk. These results suggest that this treatment program may effectively prevent midge outbreaks in Israel's drinking water supply system during the height of the summer.
Animals↗
Health care CBA and CEA from 1991 to 1996: an updated bibliography.
OBJECTIVES: This article updates, through 1996, a previously published bibliography of health-care cost-benefit and cost-effectiveness analysis, which described the literature from 1979 to 1990. METHODS: A systematic search of MEDLARS databases was conducted for all articles falling under the medical subject headings "cost-benefit analysis" (which includes cost-effectiveness analysis) and "costs and cost analysis," as well as any article with the term "cost" in the title or abstract. All titles and abstracts were scanned to determine whether articles pertained to personal health services and whether both costs and consequences were assessed. If both criteria were met, the article was included in the bibliography. RESULTS: This search resulted in 3,539 eligible cost-benefit/cost-effectiveness analysis publications from 1991 through 1996. Publications were subdivided into two major categories: reports of studies and "other" publications, including reviews, descriptions of methodology, letters, and editorials. Reports of studies and "other" publications were classified into approximately 250 different topic areas. Studies were further classified by parameters such as study type, publication vehicle, and medical function. This article describes the results of this classification and describes trends during 1991 to 1996 as compared with 1979 to 1990. OVERVIEW OF CONTENTS: The entire bibliography is reproduced in Appendix A. The classification of study reports and "other" publications into topic areas is presented in Appendix B, with numbered references to all bibliography entries. Detailed tables of findings are presented in Appendix C, and the results are illustrated graphically in Appendix D.
Bibliographies as Topic↗