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Biomedical subjects

V H Denenberg

Publications and source records attributed to V H Denenberg.

At least 55 records · Page 3Linked to original sources

Effects of embryo transfer and cortical ectopias upon the behavior of BXSB-Yaa and BXSB-Yaa + mice.

The BXSB-Yaa and BXSB-Yaa + inbred strains of mice differ primarily with respect to the Y chromosome, although there is evidence that they differ on several autosomal genes as well. Each strain has ectopic collections of neurons in neocortical layer I (ectopias), with a higher occurrence in males (58%) than females (42%). Conventionally reared mice from these strains were compared to mice that were transferred, as 8-cell embryos, into the uteri of non-autoimmune recipients, who gave birth to and reared the offspring. The transfer procedure did not change the incidence of ectopias in either sex. There were, however, major differences in behavior. Compared to conventionally reared controls, embryo transfer mice had greater behavioral asymmetry, poorer performance in a black-white discrimination, poorer Morris maze learning, better Lashley maze learning, and better performance in a two-way shuttlebox. Within the transfer groups, females differed as much as males, confirming our prior findings and supporting our thesis that the two strains differ on several autosomal genes in addition to the Y chromosome. These findings show that the intra-uterine environment can powerfully and selectively affect later behavior. When ectopic and non-ectopic mice were compared, BXSB-Yaa mice with neocortical ectopias were better able to learn the Morris spatial maze than non-ectopic controls; this was true whether the mice were conventionally reared or embryo transferred. In contrast, BXSB-Yaa + ectopic mice did not differ from their controls if conventionally reared, but were much worse than controls if embryo transferred.

Animals↗

A computer-aided procedure for measuring Hebb-Williams maze performance.

We describe a shortened procedure for testing mice over a 5-day interval on a swimming version of the Hebb-Williams maze. The mice are given 1 day of adaptation training, and are tested over the next 4 days on Hebb-Williams problems 1, 6, 12, and 5, in that order. As an animal swims through one of the maze problems, the computer screen shows the maze pattern, and an observer traces the path taken with a computer mouse. The computer program, Observe Software, stores the path sequence, determines the error score for that trial, and sends the information to a spreadsheet where it is available for statistical analyses.

Animals↗

Lack of activational influence of ovarian hormones on the size of the female rat's corpus callosum.

The sex difference in the midsagittal area of the adult rat corpus callosum (CC) has been shown to be mediated, in part, by gonadal steroids in early development, with the sensitive period of hormone action in the female extending at least up to postnatal day 25. Given this prolonged sensitivity, the current study attempted to delineate organizational vs. activational influences of gonadal hormones on the female rat CC. In Experiment 1, callosal size was examined across the estrous cycle at 52 and 90 days of age. In Experiment 2, females were ovariectomized at 78 days and CC parameters assessed at 110 days. Last, in Experiment 3, females were ovariectomized at 78 days and sacrificed at 110 days; in addition, sham females were sacrificed during proestrus or estrus. Neither stage of estrous cycle nor adult ovariectomy affected midsagittal CC size. These results provide evidence for organizational effects of ovarian steroids on the female callosum, with the sensitive period of hormone action ending sometime between days 25 and 78.

Animals↗

Sex differences in the distribution of axon types within the genu of the rat corpus callosum.

Neuroanatomical sex differences have been documented in the rat neocortex, including dimorphism of its predominant commissure, the corpus callosum (CC). In particular, CC sex differences have been reported in the ultrastructure of the posterior callosal region, the splenium. Since the CC is a heterogeneous fiber tract with its axons arising from distinct cortical areas and passing through restricted regions along its length, it became of interest to ascertain whether cellular sexual dimorphisms may also be present in another division of the CC. To test this hypothesis, electron microscopy was used to examine axon composition in adult male and female rats in the anterior portion, the genu. The number and size of axons, the thickness of the myelin sheath, and the area within the genu occupied by these constituents, were quantified. Results showed a significant sex difference in the ratio of unmyelinated to myelinated axons, with females having a larger proportion of unmyelinated fibers. This effect was present for both (1) the number of axons, and (2) the area taken up by axonal fibers. No differences were found in the size of either axon type, or for myelin thickness. Comparison of these results with those from the splenium and possible mechanisms underlying this dimorphism are discussed.

Analysis of Variance↗

Behavioral consequences of neonatal injury of the neocortex.

Several strains of autoimmune mice spontaneously develop molecular layer ectopias that are similar in appearance to those seen in humans and are caused by disturbances in neocortical neuronal migration. These mice also exhibit behavioral anomalies, some of which correlate with ectopias, others with the immunological disorder. In this study, we induced neocortical ectopias (via puncture wounds) and microgyria (via freezing lesions) in the neocortex of 1-day-old (newborn) mice without immune disorders in an attempt to further disentangle the effects of autoimmunity and of cortical malformation on behavior. In addition, we wished to compare the behavioral effects of small ectopias to larger microgyric lesions. DBA mice were assigned at birth to receive either a puncture wound or freezing lesion of either the left or right hemisphere. An independent group was subjected to sham surgery. In adulthood, these mice were given a battery of tests designed to measure lateralization and learning capacity. Lesioned mice (irrespective of hemisphere or type of damage) performed poorly when compared to sham-operated animals in discrimination learning, in a spatial Morris Maze Match-to-Sample task, and in a Lashley Type III maze. In shuttlebox avoidance conditioning, where immunological disorder has been shown to compromise behavioral performance in autoimmune mice, there was no difference between lesioned and sham animals. These results (1) support the dissociation between the effects of developmental neocortical anomalies and autoimmune disease on behavior (2) reveal similarities between spontaneous and induced neocortical malformations and (3) fail to support a difference in behavioral effects between ectopias and microgyria.

Animals↗

Analysis of two measures of paw preference in a large population of inbred mice.

Two measures of lateralized forelimb usage, the Collins paw preference test and the Lateral Paw Preference test (LPP), were tested in 693 mice of 29 inbred strains and F1 crosses, and 2 embryo transfer groups. These strains included NZB, SM, and the NXSM recombinant inbred (RI) strains; RF and the NXRF RI strains; BXSB and the Y-consomic BXSB-Yaa+; DBA/2 and F1s of DBA/2 and BXSB and BXSB-Yaa; and NZB x NZW F1s. The findings indicated that (1) the Collins and LPP tests were independent in terms of direction of lateralization, (2) there were significant population biases of 60.96% rightward on the LPP test, and 54.39% leftward on the Collins test, (3) there were significant strain differences on measures of absolute asymmetry on both tests, (4) there were strain differences for direction of asymmetry on the Collins test, but not on LPP, (5) on the basis of the NXSM RI strains, 3 genetic loci contribute to strength of laterality on the LPP test, and (6) there was a strong correlation among strain means for strength of lateralization on the two tests. These results extend previous findings on the strength and direction of laterality, showing that two independent systems for direction of laterality may depend on the same mechanism for magnitude, and establish that multiple factors of handedness, previously identified in humans and other primates, also exist in mice.

Animals↗

Autoimmunity and avoidance learning in NXRF recombinant inbred strains.

Four recombinant inbred (RI) strains, generated from NZB and RF progenitors, were tested on a two-way shuttlebox and their sera were assayed for serological factors. The four RI strains differed with respect to number of null responses and escape time. The strain with the poorest performance displayed the greatest degree of autoimmunity. The avoidance measures were correlated against the autoimmune serological parameters for the four RI strains and the two progenitors. The number of avoidances was negatively correlated with anti-DNA antibody titer, while the number of null responses showed a positive association with immune complex level. Escape time had positive correlations with IgG, IgM-RF, and immune complexes. These findings are consistent with, and extend, our prior reports that avoidance learning is inversely related to the degree of autoimmunity.

Animals↗

Human corpus callosum: a stable mathematical model of regional neuroanatomy.

The functional utility of regional parameters of the corpus callosum (CC) derived from factor analysis has been demonstrated in the rat and the human. In this study, we have utilized factor analysis to compare CC structure between two independent groups of human subjects. Tracings of the CC were taken from the midsagittal MRIs of two human samples consisting of 103 healthy volunteers and 146 neurological patients. CCs were digitized and factor analyses were performed on 99 widths, area, axis length, and perimeter. The factor structures of these two data sets were highly congruent except that one factor analysis yielded a seven-factor solution, whereas the factor analysis of the other subject population gave six factors. Further study determined that the six-factor structure could be attributed to the presence of non-consistent right-handed males whose wider CCs in the isthmus region biased the correlations. When CC parameters of consistent and non-consistent right-handed subjects were standardized, a seven-factor structure resulted. Reproducibility of the factor structure indicates that there are shared neuroanatomical characteristics of CC morphology among two disparate populations of humans in spite of variability in CC size due to sex, age, hand preference, and hand consistency.

Adolescent↗

Ovarian estrogen acts to feminize the female rat's corpus callosum.

The rat corpus callosum (CC) is sexually dimorphic, with the male CC being larger. Ovariectomy (Ovx) on day 12 has been shown to eliminate this sex difference, with callosal values of Ovx females approaching those of male controls. This suggested that postnatal ovarian estrogen affects the size of the female CC. In the present experiment, one group of female rats received Ovx on day 12, and a second group received Ovx followed by chronic implantation of a silastic tube containing beta-estradiol on day 25. Unmanipulated males and sham females served as controls. Examination of the CC at 110 days confirmed our prior findings that males have larger callosa than females and that the Ovx group had increased CC's compared to sham controls. Our new finding was that estrogen treatment was capable of reversing the effects of Ovx. Ovx+estrogen-treated females had decreased CC size as compared to Ovx alone. Indeed, they also had smaller CC values than control females. These findings indicate that ovarian estrogen plays a role in determining CC morphology and that estrogen in the female acts to inhibit overall callosal growth as measured by changes in gross callosal size.

Analysis of Variance↗

Behavior, cortical ectopias, and autoimmunity in BXSB-Yaa and BXSB-Yaa+ mice.

The BXSB-Yaa recombinant inbred strain was created by crossing a male SB/Le with a female C57BL/6J. A Y chromosome factor derived from the SB/Le male, known as the autoimmune accelerator (Yaa), leads to an earlier onset and greater severity of autoimmune disease in males. In contrast, male BXSB mice, which lack the Yaa gene (called BXSB-Yaa+) because their Y chromosome is derived from the C57BL/6J, do not develop an autoimmune condition. To examine the influence of the Y chromosome on behavior, cortical ectopia incidence, and immune functioning, males and females of these two strains were compared. Significant strain differences (for both sexes) were found for behavioral measures including discrimination, spatial and avoidance learning, and activity. For immunological parameters, a sex difference was seen in the BXSB-Yaa (males more autoimmune), but not in the BXSB-Yaa+ strain. As expected, male BXSB-Yaas were more autoimmune than male BXSB-Yaa+s. However, there was also a strain difference for IgG in the females (BXSB-Yaa+ greater). No strain difference was found for the presence of ectopias. However, there was a sex difference across both strains, with males having a higher incidence. BXSB-Yaa and BXSB-Yaa+ mice have behavioral and immunological differences greater than would be predicted by their known genetic differences. The significant differences between the two female groups suggest that the two strains differ with respect to autosomal genes, in addition to the Y chromosome. The incidence of ectopias is independent of this genetic difference and is influenced by the subject's sex.

Animals↗

Reliability and validity of computer scoring of behavioral sleep-wake states in rats and rabbits.

Previous studies in human infants, rabbits, and rats have shown that states of sleep and wakefulness can be reliably identified from motility signals produced by respiration and body movement. Thoman has described a computer-scoring algorithm for automated scoring of behavioral states from motility signals in human infants. In the present studies, we report the use of the computer scoring program with motility signals obtained from electronic activity monitors. In the newborn rabbit, computer classification of the data into behavioral states agreed with visual scoring of the motility signals. In infant and adult rats, computer scoring across the entire age range was validated against direct behavioral observations, video-taped observations, and visual scoring of the motility signal. This procedure permits prolonged recordings of freely moving animals and eliminates the need for an observer to be present.

Aging↗

Multiple dimensions of handedness and the human corpus callosum.

Consistency of right-hand preference correlates with callosal isthmus size. Anterior corpus callosum (CC) regions vary as a function of sex and right- versus left-hand preference. We analyzed regional CC anatomy in a sample of right- and left-handed men and women on the basis of both degree ("Consistency") and direction ("Hand") of hand preference scores. Significant Hand x Consistency effects indicated that both are important factors in CC organization. These data also suggested that a single continuum of hand preference and regional CC size may not be applicable across all subjects.

Adolescent↗

Environmental enrichment, neocortical ectopias, and behavior in the autoimmune NZB mouse.

New Zealand Black (NZB) mice have severe autoimmune disease and approximately 40% have cortical ectopias in layer I of sensorimotor cortex. Because the ectopias are similar to those found in dyslexics, NZB mice have been used as an animal model for developmental learning disorders. In addition, these mice have been used as a model of learning deficits associated with autoimmune disease. To determine whether early intervention would affect learning processes in NZB mice, they were reared after weaning in standard cages or enriched environments. They were given a battery of behavioral tests to measure learning, laterality, and activity, after which they were sacrificed and their brains examined for cortical ectopias. The tests sorted into two behavioral sets. Ectopia-associated behaviors included black-white discrimination learning and the Morris spatial maze. As a group, the mice performed well on these tasks. Ectopic mice had poorer performance than non-ectopics on these measures, and environmental enrichment countered the effects of the ectopias. Autoimmune-associated behavior involved two-way avoidance learning in a shuttlebox. Mice were uniformely poor on this task, ectopias did not affect behavior, and environmental enrichment was without benefit. Evidence from this and other studies shows that poor shuttlebox performance is related to the presence of autoimmune disease. Thus, autoimmune disease and cortical ectopias each appear to affect a separate set of behavioral processes. Environmental enrichment is most effective for behavioral impairments mediated via cortical ectopias, but is much less effective, if at all, if autoimmunity is the primary mediator of the impairments.

Animals↗

A developmental study of sex and age interactions in the human corpus callosum.

Regional analysis of the corpus callosum (CC) has demonstrated that the development of this fiber tract is sexually dimorphic. Midsagittal CC tracings from magnetic resonance images (MRIs) of 73 pairs of age-matched (age 2-79 years) males and females were digitized using computer assisted software. Area, axis length, perimeter, and 99 widths were obtained. Widths were grouped into the following regions based upon prior factor analysis: widths 3-18 (W3-18), W22-39, W49-62, W65-74, W77-85, W89-94, and W95-99. Trend analyses were performed with Sex and Age (blocked into 10-year bins) as independent variables. Higher order Sex by Age interactions were significant in regions W3-18 and W22-39 and were marginal in W95-99. In all 3 regions, females did not attain maximum width until Age 41-50 whereas males had peaked at 20 years and declined thereafter. There were no significant interactions between Sex and Age in the remaining CC regions, nor were there any dramatic decreases once maximum width was reached in adulthood. These results suggest that sex differences may also exist in CC ultrastructure over the human lifespan.

Adolescent↗

Behavior, ectopias and immunity in BD/DB reciprocal crosses.

In a previous study, in which fertilized DBA ova were transferred into an autoimmune female, and NZB ova were transferred into a non-autoimmune female, we found that (1) the maternal environment affected the degree of autoimmunity, (2) the incidence of cortical ectopias was not affected by the maternal environment (3) DBA and NZB females had greater paw asymmetry if reared in an autoimmune uterus, and (4) avoidance learning scores were inversely related to degree of autoimmunity. In the present experiment, reciprocal crosses of DBA and BXSB mice were studied to confirm and extend the original findings. DB mice (DBA female x BXSB male) had greater immune activity than the BD animals, had poorer avoidance learning, but were better on black-white discrimination learning and the Lashley III maze. The BD mice had greater paw asymmetry. Only one of 38 animals had a cortical ectopia. The results lead to the following conclusions: (1) there is an inverse relationship between amount of immune activity and active avoidance learning; (2) some uterine factor in autoimmune mice causes females to have greater paw asymmetry; (3) cortical ectopias are under genetic control; and (4) the lesser immune activity of the BD mice suggests that they developed a suppressor system following early exposure to autoimmunity in the uterine/maternal environment.

Animals↗

Lashley maze learning deficits in NZB mice.

In a prior study we found excellent Lashley III maze learning in BXSB mice and poor learning in NZB mice, despite the fact that both strains are autoimmune and develop cortical ectopias. This prompted us to examine NZB Lashley maze performance in detail, including comparisons to other strains and attempts to improve performance by giving additional trials with or without additional intramaze visual cues. In conventional Lashley testing (10 trials), RF mice (non-autoimmune and nonectopic) and BXSBs performed well in the Lashley maze. They had high learning indices and few errors. NZB mice performed poorly, with low learning indices and many errors. Even with additional trials or additional trials plus intramaze cues, NZB performance remained poor. The number of backward and forward errors stayed high; learning indices were low. Since both BXSB and NZB mice develop autoimmune disorders and cortical ectopias, it is unlikely that differential Lashley performance is the result of the presence of these phenomena. NZB mice are known to have alterations in their hippocampal morphology, and this is a possible mediator of the Lashley deficit.

Animals↗

A behavior profile of the MRL/Mp lpr/lpr mouse and its association with hydrocephalus.

Since the NZB and BXSB autoimmune mouse strains have cortical ectopias and associated behavioral changes, mice from the MRL/l (MRL/Mp lpr/lpr) autoimmune strain were given a series of behavioral tests followed by anatomical analyses of their brains and immune assays of plasma. No cortical ectopias were found, but this strain was discovered to have a high incidence of hydrocephalus: 46% had severe dilatation of the lateral and third ventricles, 30% had moderate dilatation, and 24% had minimal or no enlargement of the ventricles (total N = 57). Those mice in the severe hydrocephalus group were poorer in a nonspatial discrimination learning task, had less paw asymmetry, and were less active in a swimming task. No association was found between degree of hydrocephalus and any immune parameter.

Animals↗

Neonatal ovariectomy and pituitary-adrenal responsiveness in the adult rat.

Litters of female rat pups were handled daily from birth to weaning. When 12 days old the pups were ovariectomized or received sham surgery. At 70 days sham and ovariectomized animals were decapitated immediately or placed within a novel environment for 10, 20, or 40 min before decapitation. Basal plasma levels of androstenedione in ovariectomized animals were approximately half that of intact sham controls, and stress caused only a small rise in androstenedione in this group. Androstenedione levels in ovariectomized animals were consistently at or below the lower sensitivity of the radioimmunoassay, indicating that circulating levels of androstenedione in the female rat are primarily of ovarian origin. Basal ACTH and corticosterone levels did not differ significantly between the two groups. However, stress-induced levels of ACTH and corticosterone were significantly depressed in ovariectomized subjects compared to controls following exposure to the novel environment. Furthermore, while ACTH levels started to return to baseline by 40 min in controls, this effect was not observed in ovariectomized animals. These findings demonstrate that prepubertal removal of ovarian humoral factors can have a long-term impact on the responsiveness of the hypothalamic-pituitary-adrenal axis to stress.

Adrenal Glands↗