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Biomedical subjects

A K Salm

Publications and source records attributed to A K Salm.

At least 19 recordsLinked to original sources

Morphological plasticity in the neural circuitry responsible for seasonal breeding in the ewe.

An increase in the response of GnRH neurons to estrogen negative feedback is responsible for seasonal anestrus in the ewe, but the underlying neural mechanisms remain largely unknown. Neural plasticity may play an important role because the density of synaptic input to GnRH neurons changes with seasons. Moreover, the transition from breeding to anestrous season requires thyroid hormones, which are also required for neuronal development. In the first experiment, we examined whether the decrease in synapses on GnRH neurons is critical for the transition to anestrus by comparing synaptic input in thyroidectomized and thyroid-intact controls, using electron microscopic analysis. Thyroidectomized ewes remained in the breeding season, but the number of synaptic contacts on their GnRH cells was not different from those in thyroid-intact ewes that were anestrus. The next experiment tested whether there was a seasonal change in morphology of the A15 dopaminergic neurons that mediate estrogen negative feedback during anestrus by analyzing synapsin-positive close contacts onto A15 neurons with confocal microscopy. There was a 2-fold increase in these close contacts onto dendrites of A15 neurons in anestrus and a corresponding increase in the length of A15 dendrites at this time of year. The increase in dendritic length was blocked by thyroidectomy, but this procedure did not significantly affect synaptic input to A15 neurons. These results provide initial evidence that the seasonal change in synapses on GnRH neurons is not sufficient for the transition into anestrus but that plasticity of the A15 dopaminergic neurons mediating estrogen negative feedback may contribute to this seasonal alteration.

Anestrus↗

Low-frequency hearing loss in prenatally stressed rats.

We investigated the possibility that hearing thresholds are altered in prenatally stressed rats raised in a normal auditory environment. Pregnant dams were assigned randomly to prenatally stressed and control groups. Half of the dams were subjected to the mild stressors of handling, exposure to a novel cage and saline injection at random times during lights-on daily. The hearing thresholds of young adult male offspring were assessed by recording auditory-evoked brainstem responses to 0.5, 1, 2, 4, 8, 16, 32 and 64 kHz pure tones. The resultant audiograms showed that prenatally stressed offspring had significantly higher hearing thresholds than control animals at 1, 2 and 4 kHz (t-tests, P<0.05). The threshold shifts caused by prenatal stress averaged 7.7 dB across frequencies. We conclude that prenatal stress causes low-frequency hearing loss, possibly due to increased vulnerability to noise-induced hearing loss, accelerated cochlear degeneration and/or disrupted cochlear development.

Acoustic Stimulation↗

Perinatal corticosteroid effect on amygdala and hippocampus volume during brain development in the rat model.

Exposure of the fetus to corticosteroid during brain development has been suggested to cause permanent change in brain structure and has been associated with long term cognitive, behavioral and emotional impairment. We evaluated the effect of perinatal corticosteroid, at a dose similar to that which human fetuses are exposed, on cerebral cortex, corpus collosum, hippocampus, dentate gyrus and amygdala in a rat model. Rat pups were given betamethasone at day 1 (P1). Brain sections from the rat pups at postnatal day 45 (P45) were then analyzed. No differences were noted in the volumes of cerebral cortex, corpus collosum, hippocampus, dentate gyrus, or three nuclei of the amygdala compared to the control and sham groups. We concluded that a single course of betamethasone, at a comparable dose to that which the human fetus is exposed in clinical practice, had no effect on these regional brain volumes at this stage of development.

Amygdala↗

Prenatal stress affects the developmental trajectory of the rat amygdala.

The amygdala plays a critical role in generating the emotion of fear, and alterations in amygdala fear processing are thought to underlie the acquisition and maintenance of anxiety disorders. The prenatally stressed (PS) rat displays hormonal, behavioral and brain anatomical similarities to anxious humans and is useful to study the neurobiological underpinnings of pathological anxiety. We studied PS and control male rats at postnatal days 7 (P7), P25, P45 and P60. Using unbiased stereological analyses we examined the volumes, anterior-posterior lengths and total numbers of neurons and glia of the basolateral (BL), central (Ce) and lateral (La) amygdalar nuclei. We found prenatal stress-associated differences in the developmental trajectories of each nucleus. These were apparent in some measures as early as P7, most extensive at P25 and resolved by P45, at least as seen by Nissl staining. These changes were not a result of differential brain growth. This early divergence in developmental trajectories seen here may be the harbinger of PS rat amygdalas that ultimately function very differently in adulthood.

Amygdala↗

Early emergence of increased fearful behavior in prenatally stressed rats.

We have been studying the mildly prenatally stressed (PS) rat as a potentially useful animal model of anxiety disorders. Previously we have demonstrated that there are anatomical and biochemical alterations in the amygdalas of adult PS offspring and that these offspring show increased fearful behaviors. However, human data indicate that anxiety disorders often present first in early childhood and then persist throughout adolescence and adulthood. To determine if PS rats also model this characteristic of human anxiety disorders, here we asked whether behavioral indices of increased fear would be detectable at an early age. We tested the hypotheses that young PS rats would show increased behavioral fearfulness in response to an acute stressor and that this would increase with age. A mild prenatal stressor, consisting of removal of the dam from the home cage and administration of a subcutaneous injection of 0.1 ml of 0.9% saline daily, was administered during the last week of pregnancy. Offspring were tested in the defensive-withdrawal apparatus before and after exposure to restraint stress at 25, 45 and 60 days of age. PS animals showed increased defensive-withdrawal behavior following the stressor and were more fearful following restraint when compared to controls (CON). This was significant at P45 and increased to P60. Hence, fearful behaviors in PS rats emerge prior to sexual maturation and increase in magnitude thereafter, further validating our model as a means to investigate the underpinnings of anxiety disorders.

Animals↗

Divergent glial fibrillary acidic protein and its mRNA in the activated supraoptic nucleus.

Previous studies have shown decreased immunoreactive glial fibrillary acidic protein (GFAP) in the supraoptic nucleus (SON) when magnocellular neuroendocrine cells (MNCs) are activated by lactation or dehydration. This is thought to underlie structural plasticity of glial processes that occurs during these times. Here, we investigated how this apparent reduction in protein relates to GFAP mRNA expression in the dehydrated rat as visualized by in situ hybridization. Densitometry of silver grains in the SON revealed low levels of mRNA expression in control, 2-day dehydrated and 21-day rehydrated (R21) animals. Conversely, the SON from 7-day dehydrated (D7) subjects displayed significantly more silver grains. Thus, the pattern of GFAP mRNA expression is the inverse of what we previously observed for GFAP immunoreactivity in tissue sections of the SON. No differences in mRNA levels due to hydration state were seen in the lateral hypothalamic area, suggesting that increases in GFAP mRNA at D7 were specifically related to MNC activation. These data indicate a divergence in GFAP mRNA and protein expression in the SON.

Animals↗

Lateral amygdaloid nucleus expansion in adult rats is associated with exposure to prenatal stress.

Anxiety disorders in humans have been associated with chronic activation of the hypothalamic-pituitary-adrenal axis and changes in the volume of the amygdala. Interest in the etiology of anxiety disorders has led us and others to investigate the effects of prenatal stress on the brain development of adult male rat offspring. Prenatally stressed rats represent a promising animal model for anxiety disorders in that they have already been characterized as having both upregulated corticotropin-releasing factor (CRF) brain biochemistry and altered, more fearful, behaviors. Consistent with this, there is now evidence that prenatal stress also has an impact on the development of CRFergic neurons in the hypothalamic paraventricular nucleus and neurogenesis in the hippocampus. At this time, little information about the impact of prenatal stress on amygdala anatomy has been presented. Here we asked whether prenatal stress also has an impact on the development of the amygdala, because this structure plays a direct role in the emotions of anxiety and fear. Stereological measures of well-defined subregions of amydgdaloid nuclei revealed significantly expanded dimensions of the lateral nucleus in prenatally stressed offspring, due, in part, to more neurons and glia. These data may have direct import for the effect of adverse early life experiences and the etiology of anxiety disorders in humans. They also imply that early experiences may not be "grown out of" with development; in fact, the opposite might be true-adverse early life experiences may set developmental events into motion in the brain that last a lifetime.

Aging↗

Glial limitans elasticity subjacent to the supraoptic nucleus.

Two previous studies from our laboratory have indicated that the ventral glial limitans subjacent to the hypothalamic supraoptic nucleus (SON-VGL) undergoes a reversible thinning upon chronic activation of the magnocellular neuroendocrine cells (MNCs) of the supraoptic nucleus (SON). Numerous other studies have shown that MNC somata hypertrophy with activation. One aim of the current study was to understand better how SON-VGL thinning occurs. A second aim was to quantify overall changes of the MNC somata region due to cellular hypertrophy to compare relative changes in dimensions. Here, we undertook a light microscopic stereological investigation of the SON and the subjacent SON-VGL of Nissl stained material under basal and activated conditions. Astrocyte numbers in the underlying SON-VGL remained stable across hydration state as did the overall volume of the SON-VGL and dendritic zone reference area. How these data are consistent with our earlier observations of SON-VGL thinning was resolved by the finding of a highly significant, 30% increase in the mediolateral dimension of the SON-VGL in dehydrated rats. These observations fit well with previous work from our laboratory that demonstrates a reorientation of SON-VGL astrocytes, from vertical to horizontal, which occurs in the activated SON-VGL. We found a significant, approximately 54%, increase in the overall volume of the MNC region of the SON. No significant rostrocaudal lengthening of the SON was detected, although a trend was evident. All the observed changes reversed with rehydration. These data indicate that elasticity of the SON-VGL acts to accommodate the volume expansion of the MNCs and enables the SON-VGL to continue as an interface between the underlying cerebrospinal fluid in the subarachnoid space and the expanded SON above.

Adaptation, Physiological↗

Increased morphological diversity of microglia in the activated hypothalamic supraoptic nucleus.

Microglia are the immune cells of the CNS. In the normal adult mammalian brain, the majority of these cells is quiescent and exhibits a ramified morphology. Microglia are perhaps best known for their swift transformation to an activated ameboid morphology in response to pathological insults. Here we have observed the responsiveness of these cells to events surrounding the normal activation of neurosecretory neurons in the hypothalamic supraoptic nucleus (SON), a well studied model of structural plasticity in the CNS. Neurons in the SON were activated by substituting 2% saline for drinking water. Brain sections were collected from four experimental groups [controls (C), 2 d-dehydrated (2D), 7 d-dehydrated (D7), and 7 d-dehydrated/21 d-rehydrated animals (R21)] and stained with Isolectin-B4-HRP to visualize microglial cells. Based on morphological criteria, we quantified ramified, hypertrophied, and ameboid microglia using unbiased stereological techniques. Statistical analyses showed significant increases in the number of hypertrophied microglia in the D2 and D7 groups. Moreover, there was a significant increase in the number of ameboid microglia in the D7 group. No changes were seen across conditions in the number of ramified cells, nor did we observe any significant phenotypic changes in a control area of the cingulate gyrus. Hence, increased morphological diversity of microglia was found specifically in the SON and was reversible with the cessation of stimulation. These results indicate that phenotypic plasticity of microglia may be a feature of the normal structural remodeling that accompanies neuronal activation in addition to the activation that accompanies brain pathology.

Animals↗

Mild prenatal stress in rats is associated with enhanced conditioned fear.

We tested the hypothesis that prenatal stress would enhance conditioned fear in adult rats. Pregnant Sprague-Dawley rats were stressed by exposure to a novel environment and subcutaneous injection of saline (0.1 ml 0.9% NaCl) at random times daily from Days 14 to 21 of pregnancy. When compared to adult control (CON) male rats from unmanipulated pregnancies, adult prenatally stressed (PS) male rats showed increased freezing behavior in response to acute footshock as well as increased freezing behavior the next day in the same context, without shock delivery. In another experiment, the gestational stressor was examined for elevations in corticosterone and ACTH. At gestational days (G)15, G17, G19 and G21, maternal and fetal plasma was collected. Analysis showed elevations in corticosterone and ACTH in the PS dams when compared to the CON dams. Additionally, increased corticosterone was found in the PS fetuses when compared to the CON fetuses. Finally, some CON and PS litters were examined for alterations in length of gestation, number of pups born, bodyweight on postnatal day (P)1 and anogenital distance on P1 and differences were not found. In conclusion, our data demonstrate that a mild stressor during gestation, sufficient to raise plasma corticosterone and ACTH, is associated with enhanced conditioned fear during adulthood.

Adrenocorticotropic Hormone↗

Mechanisms of glial retraction in the hypothalamo-neurohypophysial system of the rat.

It has been known for more than twenty years that changes in glial coverage of magnocellular neurones in the hypothalamo-neurohypophysial system accompany activation of those neurones. This led to the so-called 'glial retraction hypothesis.' However, until recently, little has been established as to how this structural plasticity of astrocytes develops. This paper will explore a number of hypotheses and supporting data concerning these changes.

Animals↗

The surface density of glial fibrillary acidic protein immunopositive astrocytic processes in the rat supraoptic nucleus is reversibly altered by dehydration and rehydration.

Ultrastructural and immunohistochemical studies of the supraoptic nucleus (SON) have provided evidence that retraction and extension of astrocytic processes from between magnocellular neuroendocrine cells (MNCs) likely plays a role in the release of oxytocin, and/or vasopressin, that accompanies parturition, lactation and dehydration. The present study estimates the surface density (Sv) of glial fibrillary acidic protein (GFAP)-immunoreactivity, predominantly in astrocytic processes, in the SON of normally hydrated, dehydrated and rehydrated rats. The Sv of GFAP processes in dehydrated rats was significantly reduced compared with control levels. Rehydration returned Sv to control levels. The reversible reduction in Sv indicates that the previously observed reduction in optical density is due to a rearrangement of astrocyte processes in the SON which occurs at the same time as the selective functional activation of MNCs.

Animals↗

Dehydration-associated changes in the ventral glial limitans subjacent to the supraoptic nucleus include a reduction in the extent of the basal lamina but not astrocytic process shrinkage.

In these studies we have investigated factors that might account for two previous observations of the ventral glial limitans subjacent to the supraoptic nucleus (SON-VGL) of dehydrated rats: (1) a reversible reduction in the thickness of the SON-VGL, and (2) a reversible reorientation of VGL astrocytes. Since components of the basal lamina influence both cell viability and polarity, we used electron microscopic sterology to determine the volume fraction of basal lamina in the SON-VGL. We further made extensive measurements of astrocytic process thickness to determine if cellular shrinkage is a factor in the thinning of the SON-VGL. While we found no evidence for changes in the thickness of astrocytic processes, there was a significant and reversible reduction in the extent of the basal lamina. These data suggest that the thinning of the VGL is due to complex biochemical events and is not merely an epiphenomenon of dehydration.

Animals↗

Dehydration and rehydration selectively and reversibly alter glial fibrillary acidic protein immunoreactivity in the rat supraoptic nucleus and subjacent glial limitans.

Ultrastructural studies of the supraoptic nucleus (SON) of the hypothalamus suggest that an active retraction and extension of astrocytic processes (structural plasticity) from between magnocellular neuroendocrine neurons plays a role in the release of oxytocin, vasopressin, or both peptides that accompanies parturition, lactation, and dehydration. In support of this, Salm et al. (1985) previously demonstrated a lactation-associated reduction in immunoreactive glial fibrillary acidic protein (GFAP), an astrocyte-specific cytoskeletal constituent. To determine if similar changes occur in response to dehydration, and if they are reversible, the present study examined GFAP-immunoreactivity (IR) in the SON under various hydration states. Rats were dehydrated for 7 days by substitution of drinking water with 2% saline (n = 3), or dehydrated for 7 days followed by 7 days of rehydration (n = 3). A control group (n = 3) with free access to tap water was used for comparisons. The optical density of GFAP-IR was obtained from the SON, globus pallidus, and lateral hypothalamic regions. The areas of the ventral glial limitans subjacent to the SON (SON-VGL) and of linearly equivalent segments of glial limitans more distant from the SON were also determined. Dehydration resulted in a significant reduction in GFAP-IR in the SON compared to control and rehydrated levels. We also found that the area of the SON-VGL was significantly larger than that of linearly equivalent segments of glial limitans elsewhere and that it was significantly reduced in dehydrated rats, returning to control levels with rehydration. GFAP-IR and glial limitans thickness in regions unrelated to body fluid homeostasis lateral to the SON, overlying to dorsal cortex, and subjacent to the optic chiasm were not significantly changed by hydration state. These results are similar to the changes of GFAP-IR reported for lactating rats and provide further evidence for a role of structural plasticity of astrocytes in events surrounding the selective functional activation of local neurons.

Animals↗

Plasticity of astrocytes of the ventral glial limitans subjacent to the supraoptic nucleus.

We present evidence of gross morphological changes in astrocytes of the ventral glial limitans (VGL) associated with a well-known model of central nervous system (CNS) plasticity: the hypothalamic supraoptic nucleus (SON). Activity of SON magnocellular neuroendocrine cells (MNCs) was stimulated in experimental rats by substitution of 2% saline for drinking water for 2 or 9 days. Light microscopic measures revealed that a significant decrease in VGL thickness, by 34%, occurred with 9 days of stimulation. Astrocyte nuclei of 9-day dehydrated animals were also found to be 39% closer to the pial surface when compared with controls. Electron microscopy revealed a reorientation of individual astrocytes from a direction perpendicular (vertical) to the pial surface, to one parallel (horizontal) to this region. Vertically oriented astrocytes were found to be greater in the control group, by 49%, when compared with the 9-day dehydrated group, where cells were predominantly horizontal in orientation. Vertically oriented cells were further analyzed as to the direction of their vertical projections. Control, 2-day dehydrated and 9-day rehydrated animals, had more vertical cells which were oriented toward the pial surface when compared with 9-day dehydrated animals, where the relatively few vertically oriented astrocytes were significantly more likely to project toward the dendritic zone. In animals allowed to rehydrate for 9 days following a period of dehydration, these changes returned toward control levels. We conclude that astrocytes in vivo are capable of reversible gross morphological changes over a relatively short time.

Animals↗

Differential expression of tenascin by astrocytes associated with the supraoptic nucleus (SON) of hydrated and dehydrated adult rats.

The present study evaluated the expression of tenascin by astrocytes in the supraoptic nucleus and associated ventral glial limitans (SON-VGL) under conditions that induce reversible changes in neuronal organization (dehydration and rehydration). Immunostaining of astroglia cultured from rat neonatal SON-VGL confirmed that these cells are capable of both expressing and secreting tenascin. Observations of immunostained tissue sections from adult rats revealed tenascin immunoreactivity primarily in the VGL and dendritic zone, subjacent to SON neuronal somata. Comparison of immunostained tissues from hydrated and dehydrated animals showed an apparent decrease in the intensity of immunostaining with dehydration. Subsequent Western blots of similar tissues confirmed the presence of the 210-220-kDa tenascin protein in the SON-VGL. SON-VGL tissues from control, dehydrated, and rehydrated rats were then studied by using SDS-PAGE and quantitative gel densitometry. A consistent decrease in tenascin concentration was observed by 6 days of dehydration that, with rehydration, reversed back toward or beyond control levels. Together, these observations indicate that SON-VGL astrocytes variably express tenascin and that this protein may play a role in adult SON plasticity.

Animals↗

Astroglia proliferate in response to oxytocin and vasopressin.

The effects of the peptides oxytocin and vasopressin on the proliferation of cultured cortical and hypothalamic astroglia were assessed by two corroborative methods. Both hemocytometer cell counts, and immunocytochemistry for bromodeoxyuridine (BrdU) incorporation and glial fibrillary acidic protein (GFAP) expression indicate that oxytocin increases the rate of proliferation of both cortical and hypothalamic astroglia. While vasopressin also had an effect on cortical cells, no conclusive evidence for vasopressin affecting proliferation of hypothalamic astroglia was found.

Animals↗

Dehydration-induced proliferation of identified pituicytes in fully adult rats.

The mitotic activity astrocytes in the adult central nervous system (CNS) is of interest due to their roles in gliosis and tumorigenesis, and their potential in aiding recovery of function following injury or disease. The posterior pituitary offers a potentially powerful model to study proliferation in vivo, since its resident astrocytes, called pituicytes, have been reported to divide concurrently with hormone release from the neurosecretory terminals there. our aim in this study was to confirm and characterize this proliferative response during dehydration and rehydration in fully adult animals using contemporary techniques. Adult male rats were given 2% saline in substitution for water for 0-9 days. Proliferation of pituicytes was quantified in tissue sections triple-labeled with the proliferation marker, 5-bromodeoxyuridine (BrdU), the astrocyte marker glial fibrillary acidic protein (GFAP), and the DNA marker 4,6,diamidino-2-phenylindole, HCL(DAPI). A robust proliferative response began within three days of dehydration and continued at a constant rate thereafter. In animals allowed to rehydrate, this response continued. After 9 days of dehydration, approximately 35% of pituicytes had participated in mitosis. While cell density remained constant across conditions, a reversible increase in posterior pituitary area was seen, suggesting that some cell death also occurs simultaneously. A significant proportion of non-pituicytes also underwent similar changes. These results indicate that pituicytes in the adult posterior pituitary retain characteristics necessary for reentering the cell cycle in response to local factors present during neurosecretory activity. We hypothesize that this proliferative response is directly related to the morphological changes previously reported for these cells under activating conditions.

Animals↗