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Biomedical subjects

A J Silverman

Publications and source records attributed to A J Silverman.

At least 91 records · Page 5Linked to original sources
PubMed1987

Localization of a peptide sequence contained in the precursor to gonadotropin releasing hormone (GnRH).

The precursor protein that contains the sequence for the neurohormone gonadotropin releasing hormone (GnRH) also contains an additional fragment (amino acids 14-26, designated pHGnRH14-26) that can release luteinizing hormone (LH) and follicle stimulating hormone (FSH) in vitro. An immunocytochemical study was carried out to determine if this sequence could be found in its processed form in hypothalamic nerve terminals. In rats, ewes and rhesus monkeys pHGnRH14-26 was demonstrated in both neuronal cell bodies and in axon terminals. In mice, immunoreactivity was present in terminals only. No reaction was found in the hamster hypothalamus. Double label immunocytochemical studies for GnRH and pHGnRH14-26 showed that both sequences could be found in the rat septal-preoptic-hypothalamic continuum in some but not all neuronal cell bodies. Control experiments strongly suggest that the 14-26 immunoreactivity represents the fragment after it has been cleaved from the precursor protein and that this peptide could be available for release into the hypophysial portal system.

Amino Acid Sequence↗
PubMed1987

Female sexual behavior in hypogonadal mice with GnRH-containing brain grafts.

Hypogonadal female mice respond to GnRH-containing fetal preoptic area (POA) implants in the third ventricle with vaginal opening and persistent vaginal estrus, ovarian, and uterine development and increased gonadotropin secretion. When these females are mated with normal males, reflex ovulation results in pregnancy. In the present study, POA implants derived from neonatal pups, whether male or female, were also capable of supporting reproductive development in the hypogonadal female mice. Evaluation of female sexual behavior in the mice with grafts showed that these mice responded to normal males with comparable levels of lordosis as are seen in normal female mice on the proestrous days of their cycles.

Animals↗
PubMed1987

Implantation of fetal preoptic area into the lateral ventricle of adult hypogonadal mutant mice: the pattern of gonadotropin-releasing hormone axonal outgrowth into the host brain.

Transplantation of fetal preoptic area tissue containing gonadotropin-releasing hormone neurons into the third ventricle of male hypogonadal mice resulted in an elevation of pituitary gonadotropin levels and correction of hypogonadism. This reversal of the neuroendocrine deficit was correlated with innervation of the median eminence by gonadotropin-releasing hormone axons. The specificity of fiber outgrowth suggested that local neuromodulatory factors might guide these axons to the nearby median eminence. To test this hypothesis, 14 adult hypogonadal males received unilateral fetal preoptic area grafts into the lateral ventricle, a site distant from the median eminence. After four months, healthy grafts containing numerous gonadotropin-releasing hormone neurons were seen in 9 hosts. However, none of these grafts corrected the hypogonadism of the host and there was no gonadotropin-releasing hormone innervation of the median eminence in any of these animals, thus demonstrating that the presence of gonadotropin-releasing hormone neurons in the ventricular space is itself not sufficient to stimulate the pituitary-gonadal axis. Instead, gonadotropin-releasing hormone axons coursed in the host fimbria, fornix, corpus callosum, and stria terminalis. These fibers could be traced into the anterior hippocampal area, medial and lateral septum, and the anterior hypothalamus. The distribution of these fibers included a number of regions which receive gonadotropin-releasing hormone fiber input in the normal mouse. These findings show that gonadotropin-releasing hormone neurons transplanted into the lateral ventricle can survive and extend processes into the host brain, often projecting to sites of normal gonadotropin-releasing hormone innervation. Their success in contacting these sites suggests that gonadotropin-releasing hormone fiber outgrowth may be influenced by regionally specified trophic and/or guidance factors.

Animals↗
PubMed1987

The effects of the age of intracerebroventricular grafts of normal preoptic area tissue upon pituitary and gonadal function in hypogonadal (HPG) mice.

Hypogonadal mice are deficient in the hypothalamic gonadotrophic hormone releasing hormone and as a consequence postnatal testicular development does not occur. Grafting preoptic area tissue from normal mice directly into the hypogonadal third ventricle dramatically reverses the hypogonadism; however, the age of the grafted preoptic area tissue is crucial to the survival and function of the graft. Grafting embryonic tissue (E16-18) resulted in 69% of the hypogonadal mice increasing testis weight some sevenfold within 30 days (5.6 to 35 mg). Postnatal day 1 (P1) tissue grafts elicited a similar rise in testis weight in 77% of recipients, whereas P5 tissue was only successful in 22% of cases. In this experimental group, however, testis weight also increased sevenfold compared with hypogonadal untreated mice. Stimulation of testicular growth in the E16-18 and P1 experimental groups was accompanied by an increase in pituitary gonadotrophic hormone content. P10 tissue did not stimulate testis growth nor was pituitary gonadotrophic hormone control elevated and the majority of grafts failed to survive over the 30 day period of the experiment. The present study has shown that the age of grafted tissue is critical in the restoration of physiological function in hypogonadal mice, and that gonadotrophic hormone releasing hormone neurons from E16-18, P1 and P5 preoptic area grafts that survive the 30 day period of the experiment and whose axons reach the median eminence portal vessels are equipotent in stimulating pituitary gonadotrophin synthesis and secretion.

Age Factors↗
PubMed1987

Localization of luteinizing hormone-releasing hormone (LHRH) neurons that project to the median eminence.

The neuropeptide, luteinizing hormone-releasing hormone (LHRH), is released from nerve terminals in the median eminence and carried via the hypophysial portal system to the anterior pituitary, where it stimulates the release of gonadotropins. LHRH-containing neurons are located in many different regions of the rodent brain, including olfactory, septal, preoptic, and hypothalamic structures. Since those LHRH neurons that project to the median eminence form the final common pathway for the regulation of the pituitary/gonadal axis, we wished to determine which of these cell groups are afferent to this structure. A retrograde tracer, the lectin wheat germ agglutinin (WGA), was placed directly on the exposed surface of median eminence. Following survival times of 8-13 hr, brains were prepared for the dual immunocytochemical detection of WGA and LHRH. Approximately 50% of the LHRH neurons from the level of the septal nuclei caudalward were found to contain WGA immunoreactivity and therefore to project to the median eminence. The remaining single-labeled LHRH neurons were intermingled with the double-labeled cells. The 2 populations were not distinguishable from each other on either cytological or cytoarchitectonic criteria. Those LHRH neurons that were not retrogradely labeled following an injection of tracer into the median eminence are presumed to project to other regions of the central nervous system. We conclude that the LHRH neurons that are directly involved in the regulation of reproductive function are very heterogeneous, widely scattered in telencephalic and diencephalic regions.

Animals↗
PubMed1986

Immunocytochemical localization of luteinizing hormone-releasing hormone (LHRH) pathways in the sheep brain during anestrus and the mid-luteal phase of the estrous cycle.

The luteinizing hormone-releasing hormone (LHRH) system of the sheep brain was examined by light microscopic immunocytochemistry with thick, unembedded sections. We compared the distribution and morphology of LHRH cells and their fibers in intact and ovariectomized anestrous ewes, and in breeding season ewes during the mid-luteal phase of their estrous cycle. In all animals, a majority of LHRH neurons were found in the medial preoptic area adjacent to the organum vasculosum of the lamina terminalis. These cells formed a continuum rostrally with immunoreactive neurons in the diagonal band of Broca and medial septum and caudally with cells in the ventrolateral anterior hypothalamus and lateral hypothalamus. Relatively few cells (1-2%) were seen in the arcuate nucleus or its vicinity. Preoptic LHRH neurons project to the tubero-infundibular sulcus of the median eminence by at least two routes: a major ventrolateral projection above the optic tract in the anterior and lateral hypothalamus, and a less prominent periventricular pathway along the third ventricle. LHRH fibers were also observed in a number of extrahypothalamic regions, including the medial amygdala and the accessory olfactory bulb. Immunoreactive LHRH neurons in the sheep exhibited a complex light microscopic morphology unlike that seen in LHRH cells of any other species to date. LHRH cells with extensive, branching processes were frequently found in clusters with close somatic appositions between neighboring cells. Multiple thin protuberances emanated from the soma of many immunoreactive neurons. Immunoreactive fibers with beaded varicosities often were intimately associated with both cell bodies and their dendritic processes. Morphometric analyses revealed that preoptic LHRH neurons in three of four mid-luteal phase ewes had a shorter total dendritic length than those neurons in either intact or ovariectomized anestrous ewes, but this difference between breeding season and anestrous ewes was not statistically significant. Evidence for possible seasonal and/or steroid-induced alterations in the morphology of LHRH neurons must be documented by further studies, including immunocytochemical observations at an ultrastructural level.

Animals↗
PubMed1986

Hypothalamic localization of multiunit electrical activity associated with pulsatile LH release in the rhesus monkey.

Abrupt increases in electrical activity can be recorded from the medial basal hypothalamus of the rhesus monkey and these bursts are correlated with a pulse of LH release from the anterior pituitary. In this study we have localized the tips of the electrodes from which such electrical activity can be recorded and have attempted to correlate the placement with the presence of immunoreactive LHRH neurons and axons. Electrodes associated with bursting activity were found as far rostral as the suprachiasmatic nucleus and as far caudal as the premammillary nuclei. The majority were found in the medial basal hypothalamus including the region of the arcuate nucleus, the retrochiasmatic zone and the dorsal aspect of the median eminence. The tips of the electrodes were associated with either LHRH neurons or axon bundles. Negative electrodes (those not associated with bursting electrical activity) which did not exit through the ventral surface of the brain nor enter the ventricle were found in the same regions of the hypothalamus as positive electrodes. Although it is tempting to associate the bursting electrical activity with the LHRH neurosecretory system, such a correlation cannot yet be made.

Animals↗
PubMed1986

Comparative distribution of vasopressin and oxytocin neurons in the rat brain using a double-label procedure.

The distribution of vasopressin (VP) and oxytocin (OT) neurons in the rat supraoptic (SON), paraventricular (PVN), and accessory magnocellular (AMN) nuclei was studied by localizing both peptides on the same section with a double immunocytochemical staining procedure employing specific monoclonal antibodies (MAB). This procedure allows us to visualize the distribution of one cell type relative to the other. In the rostral SON, VP cells lie dorsal and medial to the OT cells. Near the mid-point of the nucleus along its rostral-caudal length, there is a transition zone in which the two cell types are mixed. Proceeding caudalward, the relative locations of OT and VP cells are exchanged so that most of VP cells are located in the ventral and medial sector of the nucleus, whereas the OT cells are situated dorsal and lateral. However, there is no absolute segregation of the two types of cells anywhere in the nucleus. In the anterior part of the PVN a rostral group (rPVN) of cells composed of a medial portion and a lateral wing can be recognized. Nearly all of the cells in the rPVN are oxytocin-containing. The rPVN is separated from the next group, the middle PVN (mPVN), by a cell poor zone of about 100-150 micron. The mPVN contains both OT and VP neurons. As one proceeds caudally, the OT cells extend in the rostrocaudal direction from an anterior and ventromedial location, forming a shell around a core of VP neurons. In the most caudal PVN (cPVN), a triangular cell group characterized by fusiform cells with long-beaded processes can be distinguished from the more rounded cells of the remaining PVN. Many fusiform cells in the cPVN appear to send their axons to the posterior perifornical nucleus and the nucleus of the medial forebrain bundle. Other fusiform cells of the cPVN are oriented in a rostral-caudal plane and are situated more medially in this subdivision. The dendrites of these cells project into the mPVN while their posterior processes, most of which also appear to be dendrites, project caudally along a medial route.

Animals↗
PubMed1986

Ultrastructure of gonadotropin-releasing hormone neuronal structures derived from normal fetal preoptic area and transplanted into hypogonadal mutant (hpg) mice.

The hpg mutant mouse lacks the neurohormone gonadotropin-releasing hormone (GnRH) and hence has a reproductive deficit. This deficit can be corrected by placement of normal fetal preoptic area into the third ventricle (see Krieger et al., 1985). We have now used ultrastructural immunocytochemistry to investigate the morphology of GnRH neurons in such intraventricular grafts, the routes that their axons take as they exit into the host, and the neurosecretory terminations that they make in the host median eminence. The GnRH cells in the transplant were similar in morphology to that reported for such cells in the preoptic area of other rodents. There was a large central nucleus, frequently indented and containing 1 or 2 nucleoli. The thin rim of cytoplasm was filled with rough endoplasmic reticulum, Golgi stacks, and mitochondria. Both dendritic and axonal profiles were identified, and a modest synaptic input to the former was found. Between the host and the implant a complex multilayered ependymal zone developed, and it was through this region that GnRH axons exited into the host arcuate nucleus and median eminence, usually surrounded by ependymal or glial elements. Within the median eminence, GnRH terminals were in close association with fenestrated blood vessels forming a normal neurosecretory terminus.

Animals↗
PubMed1985

The noradrenergic innervation of vasopressin neurons in the paraventricular nucleus of the hypothalamus: an ultrastructural study using radioautography and immunocytochemistry.

Immunocytochemical and radioautographic procedures were combined at the ultrastructural level to study the noradrenergic synaptic input to vasopressin neurons in selected portions of the paraventricular nucleus of the hypothalamus (PVN) of the rat. Radioactive norepinephrine (NE) was infused into the lateral ventricle or applied topically to the region of the PVN. After appropriate survival times, brain tissues were processed for ultrastructural immunocytochemical demonstration of vasopressin using a monoclonal antibody. [3H]NE varicosities were detected by electron microscopic radioautography. In the periventricular zone of the PVN, radioactive varicosities were numerous accounting for 20-30% of all nerve terminals in this zones. These NE terminals primarily innervated dendritic processes of non-vasopressinergic neurons. Although an occasional axosomatic synapse was observed, input to vasopressin positive neurons was exclusively to their dendrites. In the lateral magnocellular sub-nucleus of the PVN (designed pvl2), noradrenergic terminals were fewer in number accounting for only 1-2% of the total. These terminals were found predominately but not exclusively making axodendritic synapses onto non-vasopressin processes. In both regions, many of the radiolabeled terminals had well-defined membrane appositions with their post-synaptic partners which included a synaptic cleft and post-synaptic density of varying thickness. In both the periventricular zone and the lateral magnocellular regions, noradrenergic varicosities were seen in close proximity to numerous blood vessels.

Animals↗
PubMed1985

A combined electron microscopic HRP and immunocytochemical study of the limbic projections to rat hypothalamic nuclei containing vasopressin and oxytocin neurons.

Light microscopic studies in our laboratory have indicated that the lateral septum, amygdala, and ventral subiculum project in a perinuclear fashion to the paraventricular (PVN), supraoptic (SON), and suprachiasmatic (SCN) nuclei (Oldfield et al., '82; Silverman and Oldfield, '84). In the present paper a combined anterograde HRP and immunocytochemical procedure has been used to determine the connectivity between these limbic efferents and peptide-containing processes emanating from the above mentioned hypothalamic nuclei. Synaptic associations were found to exist between efferents from (1) the septum and both vasopressin (VP)- and oxytocin (OX)-positive dendrites derived from cells in the PVN and SON, (2) the septum and VP dendrites dorsal to the SCN, (3) the ventral subiculum and both VP and OX dendrites arising from the PVN and SON, and (iv) the amygdala and VP dendrites from the PVN. These observations help clarify an apparent discrepancy between electrophysiological data, in which limbic efferents have been shown to influence the activity of VP and OX neurons in the PVN and SON, and anatomical evidence which indicates only a perinuclear innervation from these sites not encroaching on the hypothalamic nuclei themselves. In each case the synaptic connections are made on dendrites external to the nucleus: those lateral and ventrolateral to the PVN, dorsal to the SON, and dorsal or dorsolateral to the SCN.

Animals↗
PubMed1985

Implantation of normal fetal preoptic area into hypogonadal mutant mice: temporal relationships of the growth of gonadotropin-releasing hormone neurons and the development of the pituitary/testicular axis.

Central nervous system tissue which included the preoptic area (an area rich in gonadotropin-releasing hormone neurons) was taken from normal 17-day fetal mice and transplanted into the infundibular recess of the third ventricle of the hypothalamus of 90-day male mutant hypogonadal mouse hosts that are unable to synthesize the neurohormone, gonadotropin-releasing hormone. The growth and development of gonadotropin-releasing hormone neurons and fibers in the donor and host tissue as well as recovery of the pituitary-testicular axis were followed from 10 to 120 days post-implantation. Testicular growth was evident in 94% of the hypogonadal animals within 30 days post-implantation, continued for 90 days but showed no further increase during the remainder of the experiment. Increases in seminal vesicle weight, an index of testosterone secretion, were measurable at 30 days and continued through to the end of the experiment. Pituitary concentrations of gonadotropins were doubled at 30 days over that seen in the control mutant mouse and were maintained thereafter at normal or supranormal concentrations. In contrast plasma levels of gonadotropins, although above baseline at 30 days, never reached normal circulating levels. Nevertheless, it appeared that the concentration of luteinizing hormone achieved was sufficient to initiate and maintain testicular growth and testosterone secretion for the entire duration of the experiment. Immunocytochemical analysis of brain tissue was used to determine the presence and numbers of gonadotropin-releasing hormone neurons in the transplant and the distribution of their fibers in the donor and host tissue. The numbers of immunoreactive gonadotropin-releasing hormone neurons present at the time of sacrifice ranged from 3 to 140. Fiber outgrowth from the donor cells into the host was noted as early as 10 days post-implantation and the density of outgrowth continued to increase over the course of the experiment. Positive fibers tended to accumulate over the tuberoinfundibular sulci as they do in normal animals. In those instances where the transplant was placed a long distance from the median eminence, the gonadotropin-releasing hormone axons grew on the internal surface of the third ventricle until they reached these specific exit zones. These studies indicate that in the mutant hypogonadal mouse, central nervous system transplants from normal fetal mice can maintain the function of the pituitary-gonadal axis for periods of up to 120 days post-implantation. Outgrowth of the neurosecretory fibers begins very soon after implantation and the axons tend to follow pathways seen in normal tissue.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗
PubMed1985

A light microscopic HRP study of limbic projections to the vasopressin-containing nuclear groups of the hypothalamus.

The anterograde HRP technique has been used to define the efferent projections from the lateral septum, amygdala and ventral subiculum to the anterior hypothalamus (AH) with particular attention to the paraventricular nucleus (PVN). Each limbic region was found to project to the PVN in a perinuclear fashion leaving the nucleus itself virtually devoid of HRP-labeled terminals. This projection pattern was also characteristic of the limbic innervation of the supraoptic (SON) and suprachiasmatic (SCN) nuclei. HRP injections into limbic sites has also enabled the description of both efferent and afferent projections to the remainder of the diencephalon. These results extend the observations made previously on the projections from limbic structures to the diencephalon in the rat, particularly in regard to the distinctive relationship of efferents to the PVN and SON, the major components of the hypothalamo-hypophyseal neurosecretory system. At the light microscopic level it cannot be stated with certainty whether or not such limbic afferents synapse with the dendrites of cells in the PVN, SON or SCN which extend beyond the cellular boundaries of each nucleus.

Amygdala↗
PubMed1985

Synaptic interactions of luteinizing hormone-releasing hormone (LHRH) neurons in the guinea pig preoptic area.

Previous studies from many laboratories have failed to demonstrate a significant synaptic input to luteinizing hormone-releasing hormone (LHRH) neurons in the rodent or primate hypothalamus/preoptic area. Having now developed immunocytochemical procedures that result in excellent ultrastructural preservation as well as in retention of antigenicity (Silverman AJ: J Comp Neurol 227:452, 1984), we have reinvestigated the question of the organization of the synaptic arrangements of LHRH neurons in the medial preoptic area of the guinea pig. Afferent inputs to these LHRH neurons include several varieties of axo-somatic and axo-dendritic synapses. Presynaptic terminals contain either round clear vesicles or a mixture of round and flattened vesicles. Most of these terminals, especially when serial sections are examined, contain dense-core granules. Well-defined synaptic clefts are evident and postsynaptic densities can be identified for asymmetrical connections. However, the presence of reaction product in the postsynaptic structure makes it difficult to categorize symmetrical terminals. In addition to these classical inputs, LHRH neurons also enter into complex heterodox synaptic relationships with their neighbors, including somato-dendritic and dendro-dendritic synapses in which the LHRH neuron can be either the pre- or postsynaptic element. These results suggest that complex synaptic relationships might account for the multiple levels of regulation of neurohormone release.

Animals↗
PubMed1984

Mating and pregnancy can occur in genetically hypogonadal mice with preoptic area brain grafts.

Adult female hypogonadal mice, in whom hypogonadism is secondary to a genetic deficiency in hypothalamic gonadotropin-releasing hormone (GnRH), are infertile. Mating, pregnancy, and delivery of healthy litters were achieved after transplantation of normal fetal preoptic area tissue, a major site of GnRH-containing cell bodies, into the third ventricle of adult female hypogonadal mice. Immunocytochemistry revealed GnRH-containing neurons in the grafts and GnRH-containing processes extending to the lateral median eminence of the host brains.

Animals↗
PubMed1984

Luteinizing hormone releasing hormone containing synapses in the diagonal band and preoptic area of the guinea pig.

Light microscopic immunocytochemical processing of the region of the ventral diagonal band of Broca (vertical limb) and the medial preoptic area of the guinea pig for LHRH revealed complex axonal bundles. The axons were of several different shapes and widths and showed varicosities of varying form. When such tissue was embedded and processed for electron microscopic localization of immunoreaction product, numerous LHRH-positive synapses were seen. The synapses were made by large varicosities. They were all axodendritic and showed a well-defined synaptic cleft, as well as a clustering of immunoreactive large granules and immunonegative small, clear vesicles. Postsynaptic densities were either very small or absent. These data show that LHRH is present in direct synapses within the CNS and is, therefore, in a position to be released and act as a neurotransmitter/neuromodulator as well as a neurohormone.

Animals↗