scholarly journals Morphological and biochemical studies on the development of cholinergic properties in cultured sympathetic neurons. II. Dependence on postnatal age.

1980 ◽  
Vol 84 (3) ◽  
pp. 692-704 ◽  
Author(s):  
M I Johnson ◽  
C D Ross ◽  
R P Bunge
Keyword(s):  
Synaptic Vesicle ◽  
Neuronal Development ◽  
Specific Activity ◽  
Sympathetic Neurons ◽  
Age Groups ◽  
Adult Rats ◽  
Old Rats ◽  
Cultured Sympathetic Neurons ◽  
Neurotransmitter Plasticity

Superior cervical ganglion (SCG) neurons taken from perinatal rats and dissociated in culture develop cholinergic properties. This report examines this "plasticity" of neurotransmitter function with regard to its dependence on the stage of neuronal development. Explants of SCG from rats ranging in age from 2 d to adult were cultured, and the number of neurons surviving after 6 wk in culture was evaluated. The activities of choline acetyltransferase (ChAc) and DOPA decarboxylase (DDC) were assayed for each age group over time in culture, and the cytochemistry of the synaptic vesicle population was studied after norepinephrine loading and KMnO4 fixation. The specific activity of ChAc in all explants fell during the first 3--4 d in culture (secondary to degeneration of presynaptic terminals), with an increase during the next 30 d in explants from all age groups except in those from the 22-d-old and adult rats. The highest activity found after 1 mo in culture was in explants from 2-d-old rats (62.5 mmol per kg dry wt per h); the lowest was in explants from adults (1.3 nmol per kg dry wt per h). After 1 mo in vitro, there were no significant differences in DDC activity among explants from animals of any age (similar to approximately 220 mmol per kg dry wt per h). Co-culture of the SCG explants with heart muscle increased even further the ChAc activity in explants from 2-d-old rats but not in explants from 16-d-old and 6.5-wk-old animals. The cytochemistry of the synaptic vesicle population in 1-mo-old cultures correlated well with the ChAc activity; when the ChAc activity was high, the proportion of synaptic vesicles with clear centers was 71--88%. In explants from adult animals, only 12% of the vesicles contained clear centers. From these data we conclude that the maturity of the SCG neuron influences the degree to which it is able to adjust its neurotransmitter mechanisms. That the axons of this neuron are interacting with target tissues during the time that neurotransmitter plasticity is retained suggests that interaction with the target may play a role in the determination of transmitter type.

scholarly journals In vitro analysis of the origin and maintenance of O-2Aadult progenitor cells.

1992 ◽  
Vol 116 (1) ◽  
pp. 167-176 ◽  
Author(s):  
D Wren ◽  
G Wolswijk ◽  
M Noble
Keyword(s):  
Adult Life ◽  
Cell Types ◽  
Adult Rats ◽  
Optic Nerves ◽  
Limited Life ◽  
Old Rats ◽  
Vitro Analysis

We have been studying the differing characteristics of oligodendrocyte-type-2 astrocyte (O-2A) progenitors isolated from optic nerves of perinatal and adult rats. These two cell types display striking differences in their in vitro phenotypes. In addition, the O-2Aperinatal progenitor population appears to have a limited life-span in vivo, while O-2Aadult progenitors appear to be maintained throughout life. O-2Aperinatal progenitors seem to have largely disappeared from the optic nerve by 1 mo after birth, and are not detectable in cultures derived from optic nerves of adult rats. In contrast, O-2Aadult progenitors can first be isolated from optic nerves of 7-d-old rats and are still present in optic nerves of 1-yr-old rats. These observations raise two questions: (a) From what source do O-2Aadult progenitors originate; and (b) how is the O-2Aadult progenitor population maintained in the nerve throughout life? We now provide in vitro evidence indicating that O-2Aadult progenitors are derived directly from a subpopulation of O-2Aperinatal progenitors. We also provide evidence indicating that O-2Aadult progenitors are capable of prolonged self renewal in vitro. In addition, our data suggests that the in vitro generation of oligodendrocytes from O-2Aadult progenitors occurs primarily through asymmetric division and differentiation, in contrast with the self-extinguishing pattern of symmetric division and differentiation displayed by O-2Aperinatal progenitors in vitro. We suggest that O-2Aadult progenitors express at least some properties of stem cells and thus may be able to support the generation of both differentiated progeny cells as well as their own continued replenishment throughout adult life.

Relation between dietary-induced increase of intestinal lactase activity and lactose digestion and absorption in adult rats

1984 ◽  
Vol 247 (6) ◽  
pp. G729-G735
Author(s):  
J. Leichter ◽  
T. Goda ◽  
S. D. Bhandari ◽  
S. Bustamante ◽  
O. Koldovsky
Keyword(s):  
Glucose Absorption ◽  
Diet Group ◽  
Lactase Activity ◽  
Weight Changes ◽  
Adult Rats ◽  
High Starch ◽  
Old Rats ◽  
Starch Diet

To study the relation between dietary-induced increase of intestinal lactase activity and lactose absorption, 11-wk-old rats were fed either a high-starch (70 cal%), low-fat (7 cal%) diet or a low-starch (5 cal%), high-fat (73 cal%) diet for 7 days. Food intake and body weight changes were similar in the two dietary groups. In the first experiment, lactose absorption was studied in vivo after oral administration of 600 mg lactose (10% solution in water with added [3H]PEG) to rats fasted for 16 h. Groups of rats were killed at time 0 and at 1-h intervals for the next 3 h. Lactase activity and lactose absorption were significantly higher (P less than 0.01) in the high-starch group than in the low-starch group. In the subsequent experiment, 9-wk-old rats were fed the two isocaloric diets for 3 days. By use of the everted sac technique, we have demonstrated a significantly higher absorption of monosaccharides from lactose in the high-starch diet group; also, glucose transport was higher in the high-starch diet-fed animals. When Tris, an inhibitor of lactase, was added into the mucosal fluid, absorption of lactose was abolished and no effect was seen on glucose absorption (in vivo and in vitro). In both experiments, significant linear regression was established between lactase activity and lactose absorption. Our results thus show that the increase in lactase activity, induced by feeding a high-starch diet to adult rats, is accompanied by an increased capacity to hydrolyze lactose and absorb the constituent monosaccharides.

Longitudinal force and stress of rat's esophagus: age-related changes.

1977 ◽  
Vol 232 (6) ◽  
pp. E580
Author(s):  
M P Zabinski ◽  
P Biancani
Keyword(s):  
Age Groups ◽  
Longitudinal Direction ◽  
Longitudinal Force ◽  
Active Stress ◽  
Length Relationship ◽  
Age Related ◽  
Old Rats ◽  
The Difference

Longitudinal force-length relationship of the rat esophagus was studied in vitro in three age groups: 1 mo, 3 mo, and 12 mo. The length of maximum force development (MFD) occurs at 1.4-1.5 times the in vivo length for all age groups. The active force developed at MFD increases markedly with age. The difference in the active forces in the 3-mo and 12-mo age groups is due to differences in cross section because the active stress of the esophagus in the longitudinal direction is approximately equal for the two age groups. The active stress in the 1-mo-old rats is lower than in the 3-mo-old rats, suggesting an increased contractility of the esophagus with age in this period of development.

scholarly journals Ciliary neurotrophic factor induces cholinergic differentiation of rat sympathetic neurons in culture.

1989 ◽  
Vol 108 (5) ◽  
pp. 1807-1816 ◽  
Author(s):  
S Saadat ◽  
M Sendtner ◽  
H Rohrer
Keyword(s):  
Neurotrophic Factor ◽  
Specific Activity ◽  
Sympathetic Neurons ◽  
Ciliary Neurotrophic Factor ◽  
Activity Levels ◽  
Term Survival ◽  
Selective Survival ◽  
Enzyme Molecules

Ciliary neurotrophic factor (CNTF) influences the levels of choline acetyltransferase (ChAT) and tyrosine hydroxylase (TH) in cultures of dissociated sympathetic neurons from newborn rats. In the presence of CNTF both the total and specific activity of ChAT was increased 7 d after culture by 15- and 18-fold, respectively, as compared to cultures kept in the absence of CNTF. Between 3 and 21 d in culture in the presence of CNTF the total ChAT activity increased by a factor of greater than 100. Immunotitration demonstrated that the elevated ChAT levels were due to an increased number of enzyme molecules. In contrast to the increase in ChAT levels, the total and specific activity levels of TH were decreased by 42 and 36%, respectively, after 7 d in culture. Half-maximal effects for both ChAT increase and TH decrease were obtained at CNTF concentrations of approximately 0.6 ng and maximal levels were reached at 1 ng of CNTF per milliliter of medium. The effect of CNTF on TH and ChAT levels were seen in serum-containing medium as well as in serum-free medium. CNTF was shown to have only a small effect on the long-term survival of rat sympathetic neurons. We therefore concluded that the effects of CNTF on ChAT and TH are not due to selective survival of cells that acquire cholinergic traits in vitro, but are rather due to the induction of cholinergic differentiation of noradrenergic sympathetic neurons.

Inhibition of Leydig cell activity in vivo and in vitro in hypothyroid rats

1995 ◽  
Vol 144 (2) ◽  
pp. 293-300 ◽  
Author(s):  
F F Antony ◽  
M M Aruldhas ◽  
R C R Udhayakumar ◽  
R R M Maran ◽  
P Govindarajulu
Keyword(s):  
Body Weight ◽  
Leydig Cell ◽  
Specific Activity ◽  
Cell Activity ◽  
Adult Rats ◽  
Hydroxysteroid Dehydrogenases ◽  
Serum Lh ◽  
Prepubertal Age

Abstract Leydig cell steroidogenic activity under basal and stimulated conditions was studied in hypothyroid rats. Hypothyroidism was induced at a prepubertal age (30 days postpartum) by surgical thyroidectomy, and l-thyroxine (T4) supplementation (6 μg/100 g body weight/day for 30 days) to hypothyroid rats was begun after 30 days. Hypothyroidism for 60 days reduced serum LH and FSH without affecting prolactin. Serum and intratesticular testosterone and the specific activity of Leydig cell 3β- and 17β-hydroxysteroid dehydrogenases diminished in hypothyroid rats. The stimulatory effect of LH on Leydig cell steroidogenic activity and cAMP was also adversely affected in hypothyroid rats. All these changes were reversed by T4 supplementation. The present results suggest that prepubertal hypothyroidism suppresses both basal and stimulated Leydig cell activity in adult rats. Journal of Endocrinology (1995) 144, 293–300

scholarly journals Superfluous Role of Mammalian Septins 3 and 5 in Neuronal Development and Synaptic Transmission

2008 ◽  
Vol 28 (23) ◽  
pp. 7012-7029 ◽  
Author(s):  
Christopher W. Tsang ◽  
Michael Fedchyshyn ◽  
John Harrison ◽  
Hong Xie ◽  
Jing Xue ◽  
...  
Keyword(s):  
Synaptic Transmission ◽  
Synaptic Vesicle ◽  
Hippocampal Neurons ◽  
Neuronal Development ◽  
Synapse Formation ◽  
Vesicle Traffic ◽  
Axon Development ◽  
Central Synapses ◽  
Synaptic Vesicle Fusion

ABSTRACT The septin family of GTPases, first identified for their roles in cell division, are also expressed in postmitotic tissues. SEPT3 (G-septin) and SEPT5 (CDCrel-1) are highly expressed in neurons, enriched in presynaptic terminals, and associated with synaptic vesicles. These characteristics suggest that SEPT3 or SEPT5 might be important for synapse formation, maturation, or synaptic vesicle traffic. Since Sept5 −/− mice do not show any overt neurological phenotypes, we generated Sept3 −/− and Sept3 −/− Sept5 −/− mice and found that SEPT3 and SEPT5 are not essential for development, fertility, or viability. Changes in the expression of septins were noted in the absence of SEPT3, SEPT5, and both septins. SEPT5 association with other septins in brain tissue was unaffected by the removal of SEPT3. No abnormalities were observed in the gross morphology and synapses of the hippocampus. Similarly, axon development and synapse formation were unaffected in vitro. In cultured hippocampal neurons, the size of the recycling synaptic vesicle pool was unaltered in the absence of SEPT3. Furthermore, synaptic transmission at two different central synapses was not significantly affected in Sept3 −/− Sept5 −/− mice. These results indicate that SEPT3 and SEPT5 are dispensable for neuronal development as well as for synaptic vesicle fusion and recycling.

scholarly journals Pseudorabies Virus Us9 Directs Axonal Sorting of Viral Capsids

2007 ◽  
Vol 81 (20) ◽  
pp. 11363-11371 ◽  
Author(s):  
M. G. Lyman ◽  
B. Feierbach ◽  
D. Curanovic ◽  
M. Bisher ◽  
L. W. Enquist
Keyword(s):  
Pseudorabies Virus ◽  
Fluorescent Protein ◽  
Sympathetic Neurons ◽  
Anterograde Transport ◽  
Viral Capsids ◽  
Green Fluorescent ◽  
Cultured Sympathetic Neurons ◽  
Retrograde Spread ◽  
Axonal Sorting

ABSTRACT Pseudorabies virus (PRV) mutants lacking the Us9 gene cannot spread from presynaptic to postsynaptic neurons in the rat visual system, although retrograde spread remains unaffected. We sought to recapitulate these findings in vitro using the isolator chamber system developed in our lab for analysis of the transneuronal spread of infection. The wild-type PRV Becker strain spreads efficiently to postsynaptic neurons in vitro, whereas the Us9-null strain does not. As determined by indirect immunofluorescence, the axons of Us9-null infected neurons do not contain the glycoproteins gB and gE, suggesting that their axonal sorting is dependent on Us9. Importantly, we failed to detect viral capsids in the axons of Us9-null infected neurons. We confirmed this observation by using three different techniques: by direct fluorescence of green fluorescent protein-tagged capsids; by transmission electron microscopy; and by live-cell imaging in cultured, sympathetic neurons. This finding has broad impact on two competing models for how virus particles are trafficked inside axons during anterograde transport and redefines a role for Us9 in viral sorting and transport.

AGE-RELATED CHANGES IN THE METABOLISM OF [3H]TESTOSTERONE IN VITRO BY THE EPIDIDYMIS OF THE IMMATURE RAT

1981 ◽  
Vol 91 (1) ◽  
pp. 43-51 ◽  
Author(s):  
R. G. FOLDESY ◽  
J. H. LEATHEM
Keyword(s):  
Adult Rats ◽  
Testosterone Metabolism ◽  
Pubertal Maturation ◽  
Immature Rat ◽  
Age Related ◽  
Rat Epididymis ◽  
Old Rats ◽  
Prepubertal Rats ◽  
Cauda Epididymidis

We examined the production in vitro of 5α-reduced metabolites from testosterone by the rat epididymis during pubertal maturation. Minced caput and cauda epididymides from 30-, 45-, and 55-day-old rats were incubated with [3H]testosterone for 2 h. Analysis of the radioactive metabolites revealed both similarities and differences in the metabolic patterns compared to those reported for adult rats. As in adults, 5α-dihydrotestosterone was the most abundant metabolite produced by both epididymal segments at all three ages, and it was formed in larger quantities in the caput epididymidis than in the cauda. However, [3H]testosterone metabolism by the epididymis of the immature rat was characterized by a lower formation of 5α-androstane-3α,17β-diol and higher production of 5α-androstane-3,17-dione than in adults. Production of these two metabolites by the caput region increased and decreased respectively, toward adult levels, with increasing age. In addition, the amount of [3H]testosterone metabolized was higher with tissues from prepubertal rats (30 days of age) than with those from rats 55 days of age. These data suggest that testosterone metabolism in the caput begins to change to that of the adult during the period of pubertal maturation but apparently not until later in the cauda epididymidis.

scholarly journals Gamma-tubulin distribution in the neuron: implications for the origins of neuritic microtubules.

1992 ◽  
Vol 119 (1) ◽  
pp. 171-178 ◽  
Author(s):  
P W Baas ◽  
H C Joshi
Keyword(s):  
Immunoelectron Microscopy ◽  
Neuronal Development ◽  
Sympathetic Neurons ◽  
Critical Role ◽  
Explant Cultures ◽  
Gamma Tubulin ◽  
Cultured Sympathetic Neurons ◽  
High Degree ◽  
Tubulin Content

Axons and dendrites contain dense microtubule (MT) assays that are not attached to a traditional MT nucleating structure such as the centrosome. Nevertheless, the MTs within these neurites are highly organized with respect to their polarity, and consist of a regular 13-protofilament lattice, the two known characteristics of MTs nucleated at the centrosome. These observations suggest either that axonal and dendritic MTs arise at the centrosome, or that they are nucleated locally, following a redistribution of MT nucleating material from the centrosome during neuronal development. To begin distinguishing between these possibilities, we have determined the distribution of gamma-tubulin within cultured sympathetic neurons. gamma-tubulin, a newly discovered protein which is specifically localized to the pericentriolar region of nonneuronal cells (Zheng, Y., M. K. Jung, and B. R. Oakley. 1991. Cell. 65:817-823; Stearns, T., L. Evans, and M. Kirschner. 1991. Cell. 65:825-836), has been shown to play a critical role in MT nucleation in vivo (Joshi, H. C., M. J. Palacios, L. McNamara, and D. W. Cleveland. 1992. Nature (Lond.). 356:80-83). Because the gamma-tubulin content of individual cells is extremely low, we relied principally on the high degree of resolution and sensitivity afforded by immunoelectron microscopy. Our studies reveal that, like the situation in nonneuronal cells, gamma-tubulin is restricted to the pericentriolar region of the neuron. Furthermore, serial reconstruction analyses indicate that the minus ends of MTs in both axons and dendrites are free of gamma-tubulin immunoreactivity. The absence of gamma-tubulin from the axon was confirmed by immunoblot analyses of pure axonal fractions obtained from explant cultures. The observation that gamma-tubulin is restricted to the pericentriolar region of the neuron provides compelling support for the notion that MTs destined for axons and dendrites are nucleated at the centrosome, and subsequently released for translocation into these neurites.

Sensitivity of lungs of aging Fischer 344 rats to ozone: assessment by bronchoalveolar lavage

1996 ◽  
Vol 271 (4) ◽  
pp. L555-L565 ◽  
Author(s):  
R. Vincent ◽  
D. Vu ◽  
G. Hatch ◽  
R. Poon ◽  
K. Dreher ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Biological Effects ◽  
Age Groups ◽  
Lavage Fluid ◽  
Lung Lavage ◽  
Ozone Exposure ◽  
Fischer 344 Rats ◽  
Adult Rats ◽  
Fischer 344 ◽  
Old Rats

Biological effects indicators in bronchoalveolar lavage fluid were studied in Fischer 344 rats of different ages after exposure to 0.4-0.8 ppm ozone for periods of 2-6 h on a single day or on 4 consecutive days. The magnitude of alveolar protein transudation induced by ozone was not different between age groups, but the interindividual variability of protein changes was higher in senescent (24-mo-old) rats. By comparison to juvenile (2-mo-old) and adult (9-mo-old) rats, senescent animals had higher increases of interleukin-6 (up to 10-fold higher) and N-acetyl-beta-D-glucosaminidase (NAGA; 2-fold higher) in lung lavage after ozone. Ascorbic acid was lower in lungs of senescent rats (one-half of juvenile values), and acute ozone exposure brought a further decrease in lung ascorbate. Whereas alveolar protein transudation was attenuated after ozone exposure on 4 days, persistent elevation of NAGA in senescent rats suggested only partial adaptation. Injection of endotoxin did not modify the patterns of effects. Incorporation of 18O-ozone into macrophages and surfactant was not different between age groups, indicating that the magnified biological responses in senescent rats were not dominated by differences in internal dose of ozone. The results indicate that senescent rats respond differently than juvenile and adult rats to lung injury.

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