Young Adult Rabbits Research Articles

The renal hemodynamic effects of Aspirin in newborn and young adult rabbits.

A group of neonatal (n=10) and 12-week-old (n=12) anesthetized, ventilated New Zealand white rabbits received an acute i.v. dose (40 mg/kg body weight) of acetylsalicylic acid (ASA, Aspirin). In the neonatal animals, i.v. ASA caused within 20 min a significant (P<0.01) fall in renal blood flow and glomerular filtration rate (GFR), with an equally significant (P<0.01) increase in filtration fraction and renal vascular resistance. The latter indicates greatly augmented renal vasconstriction or more precisely reduction in intrarenal vasodilatation by inhibition of vasodilatory prostaglandin (PG) synthesis. Urine volume decreased. The 12-week-old young adult animals responded in a similar, but significantly attenuated fashion. These experiments demonstrate that inhibition of PG synthesis in neonatal animals causes very rapid, reversible vasoconstriction, with a reduction in GFR. In addition, this study confirms previous observations that the renal hemodynamic response to the inhibition of PG synthesis is far more pronounced in neonatal animals than in (young) adult rabbits.

Co-involvement of mitochondria and endoplasmic reticulum in regulation of apoptosis: changes in cytochrome c, Bcl-2 and Bax in the hippocampus of aluminum-treated rabbits

Neurodegenerative diseases, including Alzheimer’s disease, are characterized by a progressive and selective loss of neurons. Apoptosis under mitochondrial control has been implicated in this neuronal death process, involving the release of cytochrome c into the cytoplasm and initiation of the apoptosis cascade. However, a growing body of evidence suggests an active role for the endoplasmic reticulum in regulating apoptosis, either independent of mitochondrial, or in concert with mitochondrial-initiated pathways. Members of the Bcl-2 family of proteins have been shown to either inhibit apoptosis, as is the case with Bcl-2, or to promote it, in the case of Bax. Investigations in our laboratory have focused on neuronal injury resulting from the intracisternal administration of aluminum maltolate to New Zealand white rabbits, an animal system relevant to a study of human disease in that it reflects many of the histological and biochemical changes associated with Alzheimer’s disease. Here we report that treatment of young adult rabbits with aluminum maltolate induces both cytochrome c translocation into brain cytosol, and caspase-3 activation. Furthermore, as assessed by Western blot analysis, these effects are accompanied by a decrease in Bcl-2 and an increase in Bax reactivity in the endoplasmic reticulum.

Effect of endogenous beta-hydroxybutyrate on glucose metabolism in the diabetic rabbit brain: a (13)C-magnetic resonance spectroscopy study of [U-(13)C]glucose metabolites.

The neurological consequences of diabetes mellitus have recently been receiving greater attention in both clinical and experimental settings. The deleterious effect of hyperglycemia and altered oxidative substrate availability on the diabetic brain is the subject of many studies. The aim of the present study was to examine the effect of the altered metabolic environment, namely, hyperglycemia and hyperketonemia, on glucose metabolism in the diabetic brain. More specifically, we examined the effect of diabetes on the glucose flux via the pyruvate dehydrogenase (PDH) and pyruvate carboxylase (PC) pathways and subsequent metabolism in the tricarboxylic acid cycles in neurons and glia. To this end, [U-(13)C]glucose was infused into the circulation of alloxan-induced diabetic young adult rabbits, and the [(13)C]glucose metabolites were subsequently studied in brain extracts by (13)C-NMR. Significantly elevated brain glucose levels were found. In the hyperketonemic rabbits, elevated cerebral levels of beta-hydroxybutyrate (beta-HBA) were found. Alterations in the labeling patterns of glutamine in the hyperketonemic group lead to the conclusion that the elevated beta-HBA levels inhibit glucose metabolism, mostly in glia. This results in accumulation of glucose in the diabetic brain. In addition, altered levels of glutamine, glutamate, and GABA were also attributed to the effect of beta-HBA on brain metabolism. The possible role of these metabolic perturbations in causing neurological damage remains to be investigated.

Experimental assessment of the revascularization of acellular human dermis for soft-tissue augmentation.

The purpose of this experimental study was to determine whether acellular human dermis was capable of complete revascularization in a subcutaneous implantation site with various placement geometries. In young adult rabbit ears, four different sheet and rolled configurations were placed and harvested after 3, 7, 14, and 28 days with silicone rubber microangiographic injections followed by histologic analysis. Revascularization of single-layer acellular human dermis occurred rapidly and was essentially complete by 14 days after surgery. No differences were observed in the ingrowth of vessels regardless of how the basement membrane was oriented. In rolled configurations, vascular ingrowth throughout the implant was slower and had not completely penetrated the grafts by 28 days after surgery at study completion. Vessel ingrowth occurred through the implant surfaces contacting the surrounding soft tissue and along the open seam of the roll. No differences were seen whether the basement membrane was oriented on either the inside or the outside of the roll. Acellular human dermis is capable of significant revascularization of its compact collagen composition in the early postoperative period. In thicker geometries, the rate and completeness of vessel ingrowth are predictably slower. Whether complete revascularization of multilayered or rolled grafts is achieved cannot be determined from this study.

Bone response to polymers based on poly-lactic acid and having different degradation times.

Authors studied two degradable and resorbable polymers derived from lactic acid: poly-L-Lactic acid (PLLA), with a relatively long time of degradation (longer than 6 months, PL10 Purac NL); poly-DL-Lactic acid (PDLLA), with a relatively short time of degradation (shorter than 6 months, PDL Purac NL). The animal species was the young adult New Zealand White rabbit. The in-vivo study was performed by implantation of small cylinders of 10 x 3 mm in size (length x diameter) in the distal metaepiphysis of the femur; 34 cylinders have been implanted. Retrievals of PLLA specimens took place at 3, 6, 9, 12 and 24 months; for PDLLA specimens at 1, 2, 4 months. Polarized light microscopy of undecalcified tissue sections was performed. The analysis for PLLA and PDLLA has shown a favorable response of bone tissue: alterations in the bone repair, growth and remodeling have not been observed. PLLA is persistent at the times studied; there is never a tight apposition between bone and PLLA implant and an intervening fibrous layer has often been observed. PDLLA is not persistent at the times studied and it degrades quite fast; bone repair of the empty implantation's hole occurs by bony growth from the endosteal trabeculae. The newly formed bone covers the hole's walls with an elongation parallel to them. For both polymers, whether the degradation is fast or slow, the material's substitution by newly formed bone never starts from the walls of the implantation hole. Only after the complete disappearance of the polymeric material newly formed bone begins to fill the hole.

Exogenous vasopressin influences intraocular pressure via the V1 receptors

Purpose. To compare central, peripheral, and ocular effects of exogenously given vasopressin on intraocular pressure (IOP) and to identify the related receptor mechanisms of action in rabbits. Methods. Young adult New Zealand albino rabbits were entrained under a daily 12-hour light and 12-hour dark cycle. In the early light period, bolus injections of vasopressin or desmopressin (a specific V 2 receptor agonist) were given either to the central nervous system (CNS) through an implanted cannula to the 3 rd ventricle or to the systemic circulation via the ear vein in conscious rabbits. Changes in IOP and pupil size were monitored for up to 6 hours and dose-response curves were generated. Effects of centrally and peripherally given vasopressin on IOP were further examined following pretreatments with a selective V 1 receptor antagonist administered into the 3 rd ventricle and into the ear vein, respectively. In order to clarify whether or not exogenously given vasopressin can alter IOP by mechanisms inside the eye, vasopressin was injected into the anterior chamber or the vitreous chamber unilaterally in conscious rabbits. Changes in IOP and pupil size were monitored. After an anterior chamber or intravitreal injection of the V 1 receptor antagonist, changes in IOP and pupil size due to an intravenous injection of vasopressin were determined to study the involvement of the related receptor mechanism. Results. A dose-dependent elevation of IOP appeared after injections of vasopressin into the 3 rd ventricle. There was no pupillary change. This IOP elevation was blocked by the pretreatment with the V 1 receptor antagonist. Following intravenous injections of vasopressin, significant reductions of IOP and pupil size occurred. These reductions were blocked by the pretreatment with the V 1 receptor antagonist. Intracerebroventricular or intravenous injection of desmopressin had no effect on IOP or pupil size. Injection of vasopressin into the anterior chamber or the vitreous chamber caused significant reductions of IOP and pupil size. Pretreatment with the V 1 receptor antagonist into the anterior chamber or the vitreous chamber prevented the reductions of IOP and pupil size following an intravenous injection of vasopressin. Conclusions. Intracerebroventricular and intravenous injections of vasopressin cause opposite effects on IOP. The central effect of vasopressin on IOP and the peripheral effects of vasopressin on IOP and pupil size are due to stimulations of the V 1 receptors. Reductions of IOP and pupil size following intravenous injections of vasopressin are at least partially due to stimulations of the V 1 receptors inside the eye.

Effect of Acute Insulin-Induced Hypoglycemia on Fetal versus Adult Brain Fuel Utilization, Assessed by 13C MRS Isotopomer Analysis of [U-13C]Glucose Metabolites

Tight glycemic control during diabetic pregnancy has been shown to significantly reduce the occurrence of congenital malformations and other effects of maternal diabetes on the offspring. However, intensive insulin therapy often causes recurring acute maternal hypoglycemia, which has been found to be harmful to the developing fetus, although the mechanisms involved are not clear. The aim of our work was to study the effect of acute insulin-induced maternal hypoglycemia on glucose metabolism in the fetal brain. To this end, near-term pregnant New Zealand rabbits were rendered hypoglycemic, and [U-¹³C]glucose was infused into maternal circulation. The metabolic fate of the ¹³C-labeled glucose was then studied in fetal brain extracts by ¹³C NMR isotopomer analysis, together with conventional biochemical assays of glucose and lactate levels in both plasma and brain. For comparison [U-¹³C]glucose was also administered to insulin-induced hypoglycemic young adult rabbits. Our results showed that while plasma glucose levels were significantly reduced (∼70%) relative to controls, no changes in cerebral glucose levels could be detected. Lactate levels were found to be significantly decreased in hypoglycemic fetal plasma and brain. No differences in lactate levels between control and hypoglycemic young rabbit plasma and brain were observed. These differences were attributed to the utilization of lactate as an energy substrate in the fetal brain, but not in the adult brain. Higher relative ¹³C enrichments of most glucose metabolites, except lactate, in the hypoglycemic fetal and young rabbit brains, observed by ¹³C NMR, stem from reduced endogenous plasma glucose pools, thereby diluting the labeled glucose to a lower extent. The relative glucose (or glucose-derived lactate) flux via the pyruvate carboxylase and pyruvate dehydrogenase pathways (PC/PDH ratio) was not altered under hypoglycemic conditions in the fetal brain for both glutamine and glutamate, but significantly increased in the adult brain for both glutamine and glutamate. The presented data indicate the ability of the fetal brain to maintain energy metabolism during acute hypoglycemia, via lactate utilization. The increase in the adult PC/PDH ratio was suggested by us to stem from increased PC activity, in order to replenish TCA cycle intermediates.

Collagen-embedded platelet-derived growth factor DNA plasmid promotes wound healing in a dermal ulcer model.

Gene therapy has shown limited efficacy for treating congenital diseases, partly due to temporary gene expression and host immune responses. Such results suggest that gene therapy is ideal for chronic wound treatment where limited duration of target gene expression is required. This study tested the wound healing effects of topically applied platelet-derived growth factor (PDGF)-A or -B chain DNA plasmids embedded within a collagen lattice. Four 6-mm dermal ulcer wounds were created in the ears of young adult New Zealand White rabbits made ischemic by division of the central and rostral arteries. Wounds were treated with lyophilized collagen containing PDGF-B DNA (1.0-3.0 mg), PDGF-A DNA (1.0 mg), irrelevant DNA (1.0 mg), or collagen alone. Wounds were dressed and harvested after 10 days for measurement of granulation tissue formation, epithelialization, and wound closure. Results were evaluated with a paired two-tailed Student t test, with P values < 0.05 considered significant. PDGF-B DNA increased new granulation tissue (NGT) formation up to 52% and epithelialization 34% compared with controls. Wound closure was increased up to threefold. At 1.0 mg DNA, PDGF-A and PDGF-B stimulated similar responses. No difference in NGT or epithelialization was seen between control groups. PDGF DNA gene therapy is effective at accelerating wound healing in ischemic dermal ulcers and provides a viable alternative to peptide growth factor therapy.

Nicotinic modulation in an animal model of a form of associative learning impaired in Alzheimer’s disease

Eyeblink classical conditioning is a widely used associative learning paradigm that has striking behavioral and neurobiological parallels between humans and other mammals. Eyeblink conditioning is impaired in older organisms, and patients with Alzheimer’s disease (AD) are impaired beyond the normal aging deficit. The cholinergic system is of demonstrated involvement in eyeblink conditioning. Blockade of nicotinic cholinergic receptors with mecamylamine prolonged acquisition of conditioned responses (CRs) in young adult rabbits, and the nicotinic agonist, GTS-21 ameliorated conditioning deficits in older rabbits. Galantamine induces allosteric modulation of nicotinic cholinergic receptors to increase acetylcholine release as well as acting as an acetylcholinesterase inhibitor. Galantamine doses of 0.0, 1.0, 2.0, 3.0, and 4.0 mg/kg were tested in ten daily sessions in 40 retired breeder rabbits (mean age=29 months) in the 750 ms delay conditioning paradigm. A dose of 3 mg/kg galantamine was effective in improving conditioning in older rabbits, enabling them to achieve learning criterion rapidly and to produce a very high percentage of CRs. Control tests of rabbits in explicitly unpaired conditions demonstrated that non-associative factors could not account for the results. The efficacy of galantamine in a learning paradigm that shows severe impairment in AD indicates that the drug may be effective as a cognition-enhancer in AD.

Increased Excitability of Aged Rabbit CA1 Neurons after Trace Eyeblink Conditioning

Cellular properties of CA1 neurons were studied in hippocampal slices 24 hr after acquisition of trace eyeblink conditioning in young adult and aging rabbits. Aging rabbits required significantly more trials than young rabbits to reach a behavioral criterion of 60% conditioned responses in an 80 trial session. Intracellular recordings revealed that CA1 neurons from aging control rabbits had significantly larger, longer lasting postburst afterhyperpolarizations (AHPs) and greater spike frequency adaptation (accommodation) relative to those from young adult control rabbits. After learning, both young and aging CA1 neurons exhibited increased postsynaptic excitability compared with their respective age-matched control rabbits (naive and rabbits that failed to learn). Thus, after learning, CA1 neurons from both age groups had reduced postburst AHPs and reduced accommodation. No learning-related differences were seen in resting membrane potential, membrane time constant, neuron input resistance, or action potential characteristics. Furthermore, comparisons between CA1 neurons from trace-conditioned aging and trace-conditioned young adult rabbits revealed no statistically significant differences in postburst AHPs or accommodation, indicating that similar levels of postsynaptic excitability were attained during successful acquisition of trace eyeblink conditioning, regardless of rabbit age. These data represent the first in vitro demonstration of learning-related excitability changes in aging rabbit CA1 neurons and provide additional evidence for involvement of changes in postsynaptic excitability of CA1 neurons in both aging and learning.

Investigation of sexual dimorphism in the rabbit masseter muscle showing different effects of androgen deprivation in adult and young adult animals

To evaluate the role played by androgens in the development and maintenance of sex differences in the proportion of muscle fibres of different phenotypes, the effects of castration in adult (>6 months old) and in young adult (2–3 months old) male rabbits was examined. Immunohistochemical methods were used to evaluate the proportion of muscle fibres containing different myosin heavy-chain isoforms in 10 different neuromuscular compartments of the masseter. In young adult animals of both sexes, the proportion of fibres of different phenotypes in different compartments was not significantly different from that of normal adult females. In animals castrated as young adults, the development of adult male phenotype proportions was completely blocked in most compartments. In animals castrated as adults, proportions were not significantly different from those of the intact males. For most masseter compartments, androgens produced permanent changes in muscle fibre phenotype during a critical period of postnatal development. However, in the posterior deep compartment, androgen deprivation in young adults had no effect on phenotype proportions, but castration of adults resulted in a striking increase in the proportion of fibres containing the IIa myosin heavy-chain isoform.

Circadian variations of prostaglandins in the rabbit aqueous humor.

Forty-eight young adult New Zealand albino rabbits were entrained to a daily 12-hr light and 12-hr dark cycle. Under a constant dark environment, rabbits were sacrificed at 4-hr intervals, beginning at 2 hr before the accustomed lights-on period. Eight rabbits were used for each of the 6 time points within 24 hr. Aqueous humor and vitreous humor samples were collected. Samples from one eye of each rabbit were assayed for prostaglandins D2, E2, and F2alpha by enzyme immunoassay. Total protein concentrations in the samples were also determined. Concentrations of prostaglandins D2 and F2alpha in the aqueous humor were found to vary in a consistent pattern. The prostaglandin F2alpha concentration increased in the early subjective light period, remained high in this period, and decreased in the subjective dark period. The prostaglandin D2 concentration was elevated only in the early subjective light period. No significant change of aqueous humor prostaglandin E2 level was found. Total protein concentration in the aqueous humor varied in a similar pattern as the aqueous humor prostaglandin F2alpha, indicating a close association between the permeability of protein across the blood-aqueous barrier and the endogenous prostaglandin F2alpha level. In the vitreous humor, concentrations of all three prostaglandin subtypes remained unchanged, as did the total protein concentration. However, vitreous humor concentrations of prostaglandin D2 and F2alpha were significantly higher than their concentrations in the aqueous humor.

Remodeling of hippocampal synapses after hippocampus-dependent associative learning

The aim of this study was to determine whether hippocampus-dependent associative learning involves changes in the number and/or structure of hippocampal synapses. A behavioral paradigm of trace eyeblink conditioning was used. Young adult rabbits were given daily 80 trial sessions to a criterion of 80% conditioned responses in a session. During each trial, the conditioned (tone) and unconditioned (corneal airpuff) stimuli were presented with a stimulus-free or trace interval of 500 msec. Control rabbits were pseudoconditioned by equal numbers of random presentations of the same stimuli. Brain tissue was taken for morphological analyses 24 hours after the last session. Synapses were examined in the stratum radiatum of hippocampal subfield CA1. Unbiased stereological methods were used to obtain estimates of the total number of synapses in this layer as well as the area of the postsynaptic density. The data showed that the total numbers of all synaptic contacts and various morphological subtypes of synapses did not change in conditioned animals. The area of the postsynaptic density, however, was significantly increased after conditioning in axospinous nonperforated synapses. This structural alteration may reflect an addition of signal transduction proteins (such as receptors and ion channels) and the transformation of postsynaptically silent synapses into functional ones. The findings of the present study indicate that cellular mechanisms of hippocampus-dependent associative learning include the remodeling of existing hippocampal synapses. Further studies examining various time points along the learning curve are necessary to clarify the issue of whether these mechanisms also involve the formation of additional synaptic contacts.

Remodeling of hippocampal synapses after hippocampus-dependent associative learning

The aim of this study was to determine whether hippocampus-dependent associative learning involves changes in the number and/or structure of hippocampal synapses. A behavioral paradigm of trace eyeblink conditioning was used. Young adult rabbits were given daily 80 trial sessions to a criterion of 80% conditioned responses in a session. During each trial, the conditioned (tone) and unconditioned (corneal airpuff) stimuli were presented with a stimulus-free or trace interval of 500 msec. Control rabbits were pseudoconditioned by equal numbers of random presentations of the same stimuli. Brain tissue was taken for morphological analyses 24 hours after the last session. Synapses were examined in the stratum radiatum of hippocampal subfield CA1. Unbiased stereological methods were used to obtain estimates of the total number of synapses in this layer as well as the area of the postsynaptic density. The data showed that the total numbers of all synaptic contacts and various morphological subtypes of synapses did not change in conditioned animals. The area of the postsynaptic density, however, was significantly increased after conditioning in axospinous nonperforated synapses. This structural alteration may reflect an addition of signal transduction proteins (such as receptors and ion channels) and the transformation of postsynaptically silent synapses into functional ones. The findings of the present study indicate that cellular mechanisms of hippocampus-dependent associative learning include the remodeling of existing hippocampal synapses. Further studies examining various time points along the learning curve are necessary to clarify the issue of whether these mechanisms also involve the formation of additional synaptic contacts. J. Comp. Neurol. 417:49–59, 2000. ©2000 Wiley-Liss, Inc.

A morphological analysis of the motor neuron degeneration and microglial reaction in acute and chronic in vivo aluminum chloride neurotoxicity

The monthly intracisternal inoculation of aluminum chloride (AlCl 3) to young adult New Zealand white rabbits induces motor neuron degeneration marked by intraneuronal neurofilamentous aggregates similar to that observed in amyotrophic lateral sclerosis (ALS). However, in contrast to ALS, this process occurs in the experimental paradigm in the absence of a glial response. In addition, whereas ALS is a fatal disorder, the cessation of aluminum exposure leads to both clinical and neuropathological recovery. Because microglia can influence neuronal regeneration, we have examined the effect of both acute and chronic aluminum exposure on microglial activation in vivo. We have studied microglial morphology in young adult New Zealand white rabbits receiving either single (1000 μg) or repeated sublethal (100 μg monthly) intracisternal inoculums of AlCl 3. In addition, rabbits receiving 1000 μg AlCl 3 inoculums were studied following an unilateral sciatic axotomy 48 h prior to the AlCl 3 exposure. Our studies demonstrate that microglial activation in vivo is inhibited by AlCl 3 exposure, and that a correlation exists between the extent of microglia suppression and the potential for recovery. This suggests that microglial activation is an important determinant of neuronal injury.

Acceleration of wound healing with topically applied deoxyribonucleosides.

We hypothesized that a topical mixture of purified deoxyribonucleosides would accelerate wound healing in an open wound model. Full-thickness 6-mm wounds were made on the ears of young adult rabbits. In some experiments, 2 of the 3 arteries in each ear were divided to induce wound ischemia. An equiweight mixture containing all 4 of the major deoxyribonucleosides (deoxyadenosine, deoxycytidine, deoxyguanosine, and thymidine), designated PN105, or other subgroups of deoxyribonucleosides, or vehicle (saline) was applied to wounds on 1 ear every 2 days, with the other ear serving as a control. Wound tissue was processed for histological examination 7 days after the initial wounding. Granulation tissue formation and epithelialization were measured in histological cross sections of wounds. Treatment of wounds with PN105 resulted in a 191% increase in total new granulation tissue (P<.05) and a higher incidence of complete wound reepithelialization (67% vs 37%; P<.05) when compared with controls, and a similar increase under ischemic conditions on day 7. Wound ischemia markedly impairs healing; PN 105 treatment resulted in a 242% increase in the amount of new granulation tissue formed by day 7 in ischemic wounds, relative to the appropriate controls (P<.05). All 4 of the major deoxyribonucleosides were required for optimum activity; mixtures with 3 or 2 were less active or inactive. Topically applied deoxyribonucleosides reproducibly accelerate wound healing in normal and ischemic wounds, and to a magnitude equivalent to that of recombinant growth factors such as platelet-derived growth factor, previously studied in this model. In view of their safety, availability, and efficacy, deoxyribonucleosides hold considerable promise for improving healing of chronic wounds.

Effect of Castration on Rabbits Housed in Littermate Pairs

Conventional solitary caging for research rabbits precludes many postural and social behaviors. Several studies that have investigated enrichment strategies for rabbit does have found intact males usually too aggressive for paired or group housing. This study investigated the possibilities that male siblings will remain compatible if never separated from birth and that castration will improve compatibility among young sibling male rabbits. As a side benefit of pair housing for all rabbits, the larger cages (modified dog cages) allowed rest and other postures not possible in conventional single cages. In addition, there was no evidence of the stereotypies observed among individually housed rabbits. Castrated, post-weanling, male, Flemish Giant x Giant Chinchilla rabbit littermates demonstrated a significantly higher level of living together compatibly than did the intact controls. All rabbits chose to rest in close proximity or contact, although this preference was stronger in castrated males. During puberty, intact males displayed more serious aggression. We conclude that young adult littermate rabbits can live compatibly as pairs and that castration improves compatibility among males. Paired rabbits exhibit many postures and behaviors that are not possible in the small, solitary cages currently listed as minimum recommendations.

Activity of hippocampal pyramidal neurons during trace eyeblink conditioning.

The responsiveness of hippocampal pyramidal neurons was examined in young adult rabbits which were trained in a hippocampally dependent long interval trace eyeblink conditioning task. The majority of recorded cells were putative pyramidal neurons from area CA1. Our analysis indicates that 57% of 93 neurons had a statistically significant change in firing rate during some portion of the trial. The response patterns were quite heterogeneous, and inhibition was frequently observed. Inhibition was especially prominent in a post-trial period. Only a small percentage of the putative pyramidal cells exhibited robust changes in activity with prominent temporal modeling of the conditioned eyeblink response as previously described for rabbits trained with delay conditioning procedures (Berger et al. [1983] J Neurophysiol 50:1197-1219). These results were also similar for the CA3 pyramidal neurons that were recorded (N = 19). Few CA1 neurons had significant responses during unpaired presentations of tone and airpuff, and neural activity that was present was concentrated in the period commencing from airpuff onset. Preliminary data were also recorded from putative pyramidal neurons during hippocampally independent delay eyeblink conditioning, where a longer tone overlapped and coterminated with the airpuff. In this task five of 13 neurons (CA1 and CA3) were active during the trial period, another four were responsive only during the post-trial period, and other simultaneously recorded neurons were unresponsive. These data suggest a relatively more varied pattern of pyramidal neuron engagement during hippocampally dependent trace eyeblink conditioning than was apparent in previous multiple- and single-neuron analyses of activity during eyeblink conditioning.

Age-related decrease in the overall extent of perikaryal projections in rabbit spinal ganglion neurons

The overall extent of the perikaryal projections of sensory neurons from spinal ganglia of young adult and aged rabbits was estimated by a stereological method using the electron microscope. The extent of perikaryal projections was significantly smaller in the aged animals. This age-related decrease did not seem to depend on factors intrinsic to the neuron, but on the absence of a satellite cell covering over extensive portions of the nerve cell body surface. This decrease may influence the organization of the subplasmalemmal cytoskeleton, metabolic exchange between the nerve cell body and its environment and perhaps also neuronal metabolism.

Radioactive fucose as a tool for studying glycoprotein secretion.

The efficiency and reliability of radioactive fucose as a specific label for newly synthesized glycoproteins were investigated. Young adult male rabbits were injected intravitreally with [3H]-fucose, [3H]-galactose. [3H]-mannose, N-acetyl-[3H]-glucosamine or N-acetyl-[3H]-mannosamine, and killed 40 h after injection. In another series of experiments rabbits were injected with either [3H]-fucose or several tritiated amino acids and the specific activity of the vitreous proteins was determined. Vitreous samples were also processed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and histological sections of retina, ciliary body and lens (the eye components around the vitreous body) were processed for radioautography. The specific activity (counts per minute per microgram of protein) of the glycoproteins labeled with [3H]-fucose was always much higher than that of the proteins labeled with any of the other monosaccharides or any of the amino acids. There was a good correlation between the specific activity of the proteins labeled by any of the above precursors and the density of the vitreous protein bands detected by fluorography. This was also true for the silver grain density on the radioautographs of the histological sections of retina, ciliary body and lens. The contribution of radioautography (after [3H]-fucose administration) to the elucidation of the biogenesis of lysosomal and membrane glycoproteins and to the determination of the intracellular process of protein secretion was reviewed. Radioactive fucose is the precursor of choice for studying glycoprotein secretion because it is specific, efficient and practical for this purpose.