Ronald Goldstein - Academia.edu (original) (raw)

Papers by Ronald Goldstein

Journal of Virology, 2011

We also demonstrated, for the first time, VZV infection of axons and retrograde transport from ax... more We also demonstrated, for the first time, VZV infection of axons and retrograde transport from axons to neuronal cell bodies using compartmented microfluidic chambers. The use of hESC-derived human neurons in conjunction with fluorescently tagged VZV shows great promise for the study of VZV neuronal infection and axonal transport and has potential for the establishment of a model for VZV latency in human neurons.

Journal of NeuroVirology, 2012

Retrograde axonal transport of the neurotropic alphaherpesvirus Varicella zoster virus (VZV) from... more Retrograde axonal transport of the neurotropic alphaherpesvirus Varicella zoster virus (VZV) from vesicles at the skin results in sensory neuron infection and establishment of latency. Reactivation from latency leads to painful herpes zoster. The lack of a suitable animal model of these processes for the highly human-restricted VZV has resulted in a dearth of knowledge regarding the axonal transport of VZV. We recently demonstrated VZV infection of distal axons, leading to subsequent capsid transport to the neuronal somata, and replication and release of infectious virus using a new model based on neurons derived from human embryonic stem cells (hESC). In the present study, we perform a kinetic analysis of the retrograde transport of green fluorescent protein-tagged ORF23 in VZV capsids using hESC-derived neurons compartmentalized microfluidic chambers and time-lapse video microscopy. The motion of the VZV was discontinuous, showing net retrograde movement with numerous short pauses and reversals in direction. Velocities measured were higher 1 h after infection than 6 h after infection, while run lengths were similar at both time points. The hESC-derived neuron model was also used to show that reduced neuronal spread by a VZV loss-of-function mutant for ORF7 is not due to the prevention of axonal infection and transport of the virus to the neuronal somata. hESC-derived neurons are, therefore, a powerful model for studying axonal transport of VZV and molecular characteristics of neuronal infection.

The ontogenesis ofneurophysin-containing neurons in the mouse hypothalamus. PEPTIDES 1: Suppl. !,... more The ontogenesis ofneurophysin-containing neurons in the mouse hypothalamus. PEPTIDES 1: Suppl. !, 27-44, 1980.--Several aspects of the ontogeny of the neurophysincontaining neurons of the mouse hypothalamus were studied. The events investigated included: (1) the onset of appearance of neurophysin (NP) immunoreactivity in supraoptic (SON), paraventricular (PVN) and suprachiasmatic (SCN) neurons;

Vaccine, 2007

Transcutaneous immunization aims at taking advantage of the skin's immune system for the purpose ... more Transcutaneous immunization aims at taking advantage of the skin's immune system for the purpose of immunoprotection. In the present study we evaluated the potential of topical delivery of a recombinant melanoma protein, HR-gp100, derived from a shorter sequence of the native gp100 gene. The protein was applied on the skin, with and without the addition of two forms of heat labile enterotoxin (nLT and LTB). HR-gp100 fused to Haptide, a cell penetrating 20mer peptide (HR-gp100H) was also tested. Topical HR-gp100 and HR-gp100H application on the ears of mice elicited the production of specific antibodies, and transcutaneous delivery to intact human skin induced dosedependent LC activation. nLT and LTB also activated LC, but did not further increase the activation induced by HR-gp100. These results show that HR-gp100, an antigenic tumor-derived protein, activates the immune system following transcutaneous delivery, as shown by both Langerhans cell activation and induction of antibody production.

Stem Cells, 2005

have been produced from hESCs. To date, however, there have been no definitive reports of generat... more have been produced from hESCs. To date, however, there have been no definitive reports of generation of peripheral neurons from hESCs. We used a modification of the method of Sasai and colleagues for mouse and primate embryonic stem cells to elicit neuronal differentiation from hESCs. When hESCs are cocultured with the mouse stromal line PA6 for 3 weeks, neurons are induced that coexpress (a) peripherin and Brn3a, and (b) peripherin and tyrosine hydroxylase, combinations characteristic of peripheral sensory and sympathetic neurons, respectively. In vivo, peripheral sensory and sympathetic neurons develop from the neural crest (NC). Analysis of expression of mRNAs identified in other species as NC markers reveals that the PA6 cells induce NC-like cells before neuronal differentiation takes place. Several NC markers, including SNAIL, dHAND, and Sox9, are increased at 1 week of coculture relative to naive cells. Furthermore, the expression of several NC marker genes known to be downregulated upon in vivo differentiation of NC derivatives, was observed to be present at lower levels at 3 weeks of PA6-hESC coculture than at 1 week. Our report is the first on the expression of molecular markers of NC-like cells in primates, in general, and in humans, specifically. Our results suggest that this system can be used for studying molecular and cellular events in the almost inaccessible human NC, as well as for producing normal human peripheral neurons for developing therapies for diseases such as familial dysautonomia. Stem Cells 2005;23:923-930

Journal of Neurobiology, 1989

We have modified the formaldehyde-glutaraldehyde (FaGlu) histofluorescence method of Furness, Cos... more We have modified the formaldehyde-glutaraldehyde (FaGlu) histofluorescence method of Furness, Costa, and Blessing (1977a) and Furness, Costa, and Wilson (1977b) to examine wholemounts and sections of both juvenile and adult ganglia as well as peripheral tissues of Aplysia californica. FaGlu fluorescence is the result of a reaction between formaldehyde and tissue catecholamines to produce water-insoluble (fixed) fluorophores. In serially sectioned cerebral ganglia, 70-80 positive neurons were observed (many in clusters of 10-20 cells), many more than were found using the glyoxylic acid technique. Catecholamine-containing varicosities were densely packed in localized portions of the neuropil of all central ganglia. Exclusive localization in the neuropil of presumed dopamine release sites is similar to that previously found for the neuropeptide SCP but differs from the widespread ramification of varicose neurites containing 5-HT, FMRFamide, and ELH. The FaGlu technique also enabled us to study the ultrastructure of catecholamine-containing neurons. In contrast to the larger vesicles found in serotonergic and histaminergic neurons, these dopaminergic neurons contain 70 nm dense-cored vesicles.

Brain Research, 2000

Estrogen receptors (ER) are widely distributed in the central nervous system (CNS). Recent studie... more Estrogen receptors (ER) are widely distributed in the central nervous system (CNS). Recent studies, to date in rat only, have shown that ER are also expressed in neurons of the dorsal root ganglia (DRG) where they appear to have functional roles. However, no data yet exists about estrogen receptors in the embryonic DRG. In the present study, immunocytochemical staining for

Developmental Biology, 1996

The dorsal root ganglia (DRG, spinal ganglia) are a metameric series of structures that develop f... more The dorsal root ganglia (DRG, spinal ganglia) are a metameric series of structures that develop from neural crest cells within the dorsal somitic mesoderm. A striking element of patterning within this meristic series is the disappearance of the five or six most rostral DRG early in the embryonic development of birds and mammals. The transient DRG have been named “Froriep's

Developmental Biology, 1995

We have recently shown that there is very early variation in dorsal root ganglia (DRG) size based... more We have recently shown that there is very early variation in dorsal root ganglia (DRG) size based on their axial position. From the time of their condensation at Stage (St.) 20 (Embryonic Day 3), before the onset of apoptosis, the ganglia in brachial segments 14 and 15 are more than 80% larger on the average than those in cervical 5

Developmental Brain Research, 2000

The Hu and Islet-1 proteins are early markers of neuronal differentiation in the avian embryo. We... more The Hu and Islet-1 proteins are early markers of neuronal differentiation in the avian embryo. We here examine which of these markers is expressed first in avian dorsal root ganglia (DRG). Recently we showed that neural tube and DRG cells express Islet-1 after leaving the cell cycle, while sympathetic ganglion cells express Islet-1 while still dividing. Others have shown that

Developmental Biology, 1999

Classic studies have shown that the presomitic mesoderm is already committed to a specific morpho... more Classic studies have shown that the presomitic mesoderm is already committed to a specific morphological fate, for example, the ability to generate a rib. Hox gene expression in the paraxial mesoderm has also been shown to be fixed early and not susceptible to modulation by an ectopic environment. This is in contrast to the plasticity of Hox expression in neuroectodermal

Developmental Brain Research, 1999

The Islet-1 antigen is an early marker of differentiation of neural tube cells, and is expressed ... more The Islet-1 antigen is an early marker of differentiation of neural tube cells, and is expressed in many other embryonic cells as well. It had been reported that Islet-1 is expressed only in post-mitotic sympathetic neuroblasts in vitro, unlike other differentiation markers. We have double-labeled St. 23 chick embryos for bromodeoxyuridine (BrDU) and Islet-1 and found that neural tube and

PLoS pathogens, 2015

Varicella zoster virus (VZV) latency in sensory and autonomic neurons has remained enigmatic and ... more Varicella zoster virus (VZV) latency in sensory and autonomic neurons has remained enigmatic and difficult to study, and experimental reactivation has not yet been achieved. We have previously shown that human embryonic stem cell (hESC)-derived neurons are permissive to a productive and spreading VZV infection. We now demonstrate that hESC-derived neurons can also host a persistent non-productive infection lasting for weeks which can subsequently be reactivated by multiple experimental stimuli. Quiescent infections were established by exposing neurons to low titer cell-free VZV either by using acyclovir or by infection of axons in compartmented microfluidic chambers without acyclovir. VZV DNA and low levels of viral transcription were detectable by qPCR for up to seven weeks. Quiescently-infected human neuronal cultures were induced to undergo renewed viral gene and protein expression by growth factor removal or by inhibition of PI3-Kinase activity. Strikingly, incubation of culture...

Stem Cell Reviews and Reports, 2010

Xenografting of normal and transformed mammalian tissues and cells to chick embryos has been perf... more Xenografting of normal and transformed mammalian tissues and cells to chick embryos has been performed for almost 100 years. Embryonic stem cells, derived more than 25 years ago from murine, and more than 10 years ago from human blastocysts, have transformed many fields of biological research. There is a growing body of studies combining these two widely-used experimental systems. This review surveys those reports in which murine or human embryonic stem cells, or differentiated derivatives of these pluripotent stem cells, were transplanted to embryonated chick eggs. Many of these studies have utilized the unique characteristics of both experimental models to obtain answers to developmental questions that are difficult or impossible to approach with xenografting to adult rodents or tissue culture-only techniques.

Leukemia Research, 2009

Xenografting of human blood malignancies to immunodeficient SCID mice is a powerful research tool... more Xenografting of human blood malignancies to immunodeficient SCID mice is a powerful research tool. We evaluate here whether the immunodeficient turkey embryo can also serve as a xenograft host for human blood malignancies. Human leukemia, lymphoma and myeloma lines engrafted robustly into medullary and extramedullary tissues of turkey embryos as detected by PCR, FACS and histology in 8-10 days. Four of eleven patient AML samples also engrafted the bone marrow. Grafts of two lines responded to chemotherapy with doxorubicin. The turkey embryo therefore has the potential to be a complementary xenograft model for the study of human blood malignancies.

Journal of Comparative Physiology A-neuroethology Sensory Neural and Behavioral Physiology, 1991

1.The escape behavior of the cockroach, Periplaneta americana, is known to be modulated under var... more 1.The escape behavior of the cockroach, Periplaneta americana, is known to be modulated under various behavioral conditions (Camhi and Volman 1978; Camhi and Nolen 1981; Camhi 1988). Some of these modulatory effects occur in the last abdominal ganglion (Daley and Delcomyn 1981a, b; Libersat et al. 1989) and others in the thoracic ganglia (Camhi 1988). Neuromodulator substances are known to

Food-induced arousal in Aplysia is characterized by a progressive increase in the speed and stren... more Food-induced arousal in Aplysia is characterized by a progressive increase in the speed and strength of biting responses elicited by a seaweed stimulus. Data from semi- intact and dissected preparations suggest that the identi- fied, serotonergic, metacerebral cells (MCCs) of the cerebral ganglion contribute to food-induced arousal by enhancing the strength of buccal muscle contractions, and by modu- lating the

PloS one, 2015

Varicella Zoster Virus (VZV), the alphaherpesvirus that causes varicella upon primary infection a... more Varicella Zoster Virus (VZV), the alphaherpesvirus that causes varicella upon primary infection and Herpes zoster (shingles) following reactivation in latently infected neurons, is known to be fusogenic. It forms polynuclear syncytia in culture, in varicella skin lesions and in infected fetal human ganglia xenografted to mice. After axonal infection using VZV expressing green fluorescent protein (GFP) in compartmentalized microfluidic cultures there is diffuse filling of axons with GFP as well as punctate fluorescence corresponding to capsids. Use of viruses with fluorescent fusions to VZV proteins reveals that both proteins encoded by VZV genes and those of the infecting cell are transferred in bulk from infecting non-neuronal cells to axons. Similar transfer of protein to axons was observed following cell associated HSV1 infection. Fluorescence recovery after photobleaching (FRAP) experiments provide evidence that this transfer is by diffusion of proteins from the infecting cells ...

$Research paper thumbnail of Laboratory preparation of Varicella-Zoster Virus: concentration of virus-containing supernatant, use of a debris fraction and magnetofection for consistent cell-free VZV infections$

Journal of virological methods, 2014

The research laboratory generation of free Varicella-Zoster Virus (VZV) from cultured yields resu... more The research laboratory generation of free Varicella-Zoster Virus (VZV) from cultured yields results relatively low titers, with the result that most study of VZV infection utilizes cell-associated infection. However, important aspects of VZV-cell interaction, such as the entry mechanism and superinfection exclusion have not yet been studied in detail, in part due to the difficulty in obtaining a high titer cell free virus. Here, a method to generate relatively high-titer cell-free VZV, based on a combination of previously published techniques and subsequent concentration is described. VZV-infected cells are disrupted, sonicated and clarified by centrifugation. The cell-free virus in the supernatant is then concentrated to yield up to 10(5)PFU/ml. The cell debris pellet, which contains up to 10(6)PFU/ml can also be used for non cell-associated infection. Magnetic nanoparticles available commercially can be used to further enhance infection by cell-free-VZV. The tools described here ...

Methods in molecular biology (Clifton, N.J.), 2010

Peripheral somatic sensory neurons (PSNs) are responsible for the critical function of transmitti... more Peripheral somatic sensory neurons (PSNs) are responsible for the critical function of transmitting multiple modalities of information from the outside world, including heat, touch, and pain, as well as the position of muscles required for coordinated voluntary movement to the central nervous system. Many peripheral neuropathies exist, including hereditary neurodegeneration in Familial Dysautonomia, infections of PSNs by viruses such as Varicella zoster and damage to PSNs and/or their process resulting from other disease conditions such as diabetes. Understanding of the etiology of these diseases and development of treatments is hampered by the lack of normal and healthy human PSNs for study, which are only available from abortuses or rare surgical procedures.Human embryonic stem cells (hESCs) are an ideal source of cells for generating normal PSNs for study of disease and drug development, since they can be grown virtually indefinitely in tissue culture and have the potential to fo...