Immunocytochemical investigation of luteinizing hormone-releasing hormone neurons in Syrian hamsters maintained under long or short days

Light-microscope immunocytochemistry was used to investigate the LHRH system of adult male Syrian hamsters. Half of the animals were transferred from long to short photoperiods (14L:10D to 6L:18D) for 10 wk, causing plasma gonadotropin levels and the testes to revert to a prepubertal condition. In spite of the marked differences in the reproductive axis between the two groups of hamsters, the number of immunopositive LHRH neurons observed in the preoptic-medial septal area and diagonal band of Broca was approximately 400 in both cases; of these, 87-91% were monopolar and 9-13% were bipolar, regardless of whether the brains were sectioned in a coronal or sagittal plane. These results, therefore, fail to support the hypothesis that photoperiodic changes in the number of LHRH neurons play a major role in controlling the seasonal regression and recrudescence of the reproductive system in the hamster. However, morphometric analysis of the perikarya using an IBAS 2000 automatic image analyzer revealed a photoperiod-related difference. Surprisingly, the perikarya of both monopolar and bipolar LHRH neurons were significantly larger in hamsters that had been maintained on short days, as opposed to long days. These findings, therefore, are in harmony with the view that the inhibitory effect of short days on the reproductive axis is mediated through a suppression of LHRH secretion, which in turn is reflected as an increase in the net content of LHRH within the brain.     

Changes in populations of LHRH-immunopositive cell bodies following gonadectomy

One day after castration of male rats, plasma LH rose and the number of LHRH immunopositive neuronal perikarya decreased. As plasma LH continued to rise six days and three weeks post-castration, the number of LHRH immunopositive neurons also increased. The largest population of LHRH immunopositive neurons was detected three weeks post-castration and the cell group that showed the greatest increase was in the rostral preoptic area. In females, the largest population of LHRH immunopositive neurons was observed one day post-ovariectomy; at this time plasma LH levels were not significantly elevated above diestrous levels. Six days post-ovariectomy, LH levels were elevated and the number of LHRH immunopositive cells decreased. As LH levels continued to rise three weeks post-ovariectomy, the population increased in size. In males, primarily LHRH cells of the rostral preoptic area increased in in number; in females, the cell groups that increased were scattered over the diagonal band of Broca, preoptic and anterior hypothalamic areas. Although LHRH neurons demonstrated these variations following gonadectomy, there was no evidence of alteration(s) in molecular processing of precursor hormone.     

Relationship between neuropeptide Y and luteinizing-hormone-releasing hormone immunoreactivities in the hypothalamus and preoptic region

The purpose of this study was to examine the anatomical relationships of perikarya and fibers containing neuropeptide Y (NPY) and luteinizing-hormone-releasing hormone (LHRH) in the hypothalamus and preoptic region of female rats. In view of our previous report of stimulatory effects of estrogen on LHRH and NPY levels in the median eminence, animals were bilaterally ovariectomized and subsequently implanted subcutaneously with capsules containing estradiol benzoate in oil or vehicle. Following intracerebroventricular injection of colchicine, rats were perfused with fixative and their brains sectioned and processed for immunohistochemical visualization of NPY and LHRH in the same section and in consecutive sections. Estrogen treatment had no discernible effect on the distribution or relationship of these peptides. NPY-immunoreactive fibers were intimately associated with LHRH-labeled primary dendrites and perikarya in the medial preoptic region and horizontal limb of the diagonal band of Broca. Fibers containing NPY or LHRH overlapped extensively in the lateral palisade region of the median eminence and also in the subependymal and internal zones. The external zone of the median eminence displayed relatively less overlap of these peptide systems. LHRH-immunoreactive axons coursed among NPY-labeled perikarya in the arcuate nucleus and appeared to contact these cells. These results suggest that NPY-containing axons may influence LHRH-positive neurons at the cell body and also at the site of axon termination in the median eminence. LHRH-containing axons appear to contact NPY-immunoreactive perikarya in the arcuate nucleus and may interact with terminals in the median eminence. This arrangement may provide a mechanism for communication between NPY and LHRH neurons and for the neuroendocrine coordination of hypothalamic NPY and LHRH secretion before ovulation.     

Neuroendocrine changes in male hamsters following photostimulation

Transfer of gonadally regressed male golden hamsters from a short (5 L:19 D) to a stimulatory (14 L:10 D) photoperiod elicits, within 24 hr, significant changes in hypothalamic dopamine, serotonin, and possibly norepinephrine metabolism. Hypothalamic LHRH content was significantly elevated in short-photoperiod animals, but within 24 hr of transfer to a 14:10 photoperiod, LHRH declined to levels not different from those in hamsters maintained continuously in a long photoperiod. Plasma FSH levels were also significantly elevated within 24 hr of transfer, but increases in plasma LH were somewhat slower. Chronic treatment with the tyrosine hydroxylase inhibitor, alpha-methyl tyrosine (alpha MPT), which inhibits catecholamine synthesis, blocked the effect of a stimulatory photoperiod on plasma FSH levels, while treatment of 5:19 hamsters with the catecholamine precursor, L-dopa, mimicked the effects of photostimulation on plasma FSH levels. Testicular weights were not affected by alpha MPT or L-dopa treatment for 1 week. From these data, it appears that endocrine events associated with photoperiod-induced testicular recrudescence are under the control of hypothalamic neurotransmitters.     

A comparison of neuropeptide immunocytochemistry in fluid-fixed and freeze-dried brains

Summary Immunocytochemical staining of luteinizing hormone-releasing hormone (LHRH), somatostatin, and neurophysin was compared in rat brains fixed with 1) formalin, 2) Bouin's solution, 3) freeze-dried (FD), or 4) freeze-dried + paraformaldehyde vapor perfused (FDV). The distribution of LHRH fibers was similar in all preparations; however, beads of granular reaction product often appeared finer and more numerous in the median eminence of FD- and FDV brains. Positively stained LHRH perikarya were not observed in any of the preparations. In contrast, somatostatin-immunoreactive perikarya were present in the fluid-fixed and FD brains, although few were observed in FDV brains. Somatostatin-immunoreactive fibers were present in all preparations, but appeared most numerous in the median eminence of FD brains. Staining of neurophysin-containing perikarya and fibers was similar in all preparations. These observations suggest that the FD brain can provide a suitable tissue substrate for immunocytochemistry, demonstrating staining comparable to or surpassing that of more conventional preparations. However, staining of antigens in FD brain was not uniformly successful and may depend on stereochemical characteristics of each antigen as well as properties of the primary antisera used in the staining procedure.     

Localization of luteinizing hormone-releasing hormone in the forebrain of the pig

The distribution of luteinizing hormone-releasing hormone (LHRH)-immunostained perikarya and processes was examined in the forebrains of six sexually mature female pigs by use of indirect biotin-avidin horseradish peroxidase immunocytochemistry. Two primary antisera (Drs. Y.F. Chen and V.D. Ramirez CRR11B73 and Miles-Yeda UZ-4) yielded positive staining. Adjacent sections treated either primary antiserum preabsorbed with LHRH or with normal rabbit serum substituted for primary antiserum lacked positive staining. The greatest proportion of LHRH-immunostained perikarya were found in the medial preoptic area adjacent to the organum vasculosum of the lamina terminalis. The LHRH-immunostained perikarya were also scattered rostrally in the diagonal band of Broca, and within the lateral hypothalamic area, paraventricular nucleus, periventricular zone, suprachiasmatic nucleus, and medial basal hypothalamus. LHRH-immunostained processes, which extended from the medial preoptic area, coursed either along the ventral surface to the median eminence or medially and ventrally along the third ventricular wall ventrally to the median eminence and caudally to the level of the mammillary bodies. Extrahypothalamic processes were located adjacent to the lateral ventricular floor and the third ventricle from the lateral septal area (stria terminalis) to the level of the habenular nucleus. LHRH-immunostained neurons were unipolar, bipolar, and multipolar. Close associations between individual LHRH-immunostained neurons were observed.     

Electron microscopic study of immunoreactive LHRH perikarya with special reference to neuronal regulation

Summary In early postnatal rats, immunoreactive LHRH perikarya in the preoptic area were studied by light and electron microscopy. Synaptic junctions were found between the immunoreactive perikaryon or its process, and the immunonegative nerve fibers. The significance of these synapses is discussed in relation to possible mechanisms by which the activities of LHRH neurons are regulated.     

Androgen and estradiol levels in plasma and amniotic fluid of late gestational male and female hamsters: uterine position effects

Using radioimmunoassay we have measured the plasma and amniotic fluid levels of androgen and estradiol in male and female hamster fetuses nearing parturition. On Days 14 and 15 of gestation (day of birth = Day 16), plasma levels of androgen are higher in males than females while estradiol levels are equal. Amniotic fluid levels of these hormones, while lower than plasma, reflect the difference in androgen and the similarity in estradiol between sexes. Uterine position analysis on Day 14 suggests that female siblings located caudally suppress amniotic fluid androgen and elevate estradiol levels of male siblings. Comparison of Day 18 gestation male and female rat amniotic fluid androgen to Day 14 hamsters reveals that male rats are bathed in high levels of androgen. Female rats have lower levels which are not different from those of male hamsters. Female hamsters are exposed to little androgen. Relevance to behavioral sexual differentiation and the display of adult behavior is discussed.     

Effects of hypothalamic neurohormones on prolactin release from pituitary allografts in the hamster

Recent reports indicate that luteinizing hormone-releasing hormone (LHRH) releases prolactin (PRL) under some circumstances. We examined the chronic effects of LHRH, growth hormone-releasing hormone (GHRH), and corticotrophin-releasing hormone (CRH) on the release of PRL, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) by pituitary allografts in hypophysectomized, orchidectomized hamsters. Entire pituitary glands removed from 7-week-old-male Golden Syrian hamsters were placed under the renal capsule of hypophysectomized, orchidectomized 12-week-old hamsters. Beginning 6 days postgrafting, hamsters were injected subcutaneously twice daily with 1 microgram LHRH, 4 micrograms GHRH, or 4 micrograms CRH in 100 microliter of vehicle for 16 days. Six hosts from each of the four groups were decapitated on Day 17, 16 hr after the last injection. Prolactin, LH, and FSH were measured in serum collected from the trunk blood. Treatment with LHRH significantly elevated serum PRL levels above those measured in the other three groups, which were all similar to one another. Serum LH levels in hosts treated with vehicle were elevated above those measured in the other three groups. Serum FSH levels in hosts treated with LHRH were greater than FSH levels in any of the other three groups. These results indicate that chronic treatment with LHRH can stimulate PRL and FSH release by ectopic pituitary cells in the hamster.     

The role of endogenous gonadotropin-releasing hormone in the control of luteinizing hormone and testosterone secretion in the juvenile male monkey, Macaca fascicularis

To determine what changes occur in the activity of gonadotropin-releasing hormone (GnRH) neurons during pubertal development in primate species we tested the hypotheses that there are morphologic differences between GnRH-containing neurons in juvenile versus adult monkeys, and the low activity of the reproductive axis is governed by hypothalamic GnRH release in monkeys prior to puberty. We removed the brains from 5 juvenile and 5 adult male monkeys (Macaca fascicularis) and blocked, sectioned, and prepared each hypothalamus for light microscopic immunocytochemistry for GnRH-containing cells. The distribution and number of GnRH-containing neurons were similar in adult and juvenile brains; however, GnRH-containing perikarya in adult brains were significantly larger in total cross-sectional area (200 +/- 12 vs. 169 +/- 8 micron 2, P less than 0.05) and in cross-sectional area of the cytoplasm (139 +/- 2 vs. 88 +/- 6 micron 2, P less than 0.05) than in juvenile brains. In another group of 10 juvenile male macaques, we administered an antiserum to GnRH (Fraser #94; 2 ml/kg, i.v.) and monitored the effects on plasma luteinizing hormone (LH) and testosterone concentrations. The percentage of plasma samples with detectable LH levels decreased significantly (from 26.67 +/- 8.3% to 5.3 +/- 3.4%, P less than 0.05) after GnRH antiserum administration; however, plasma testosterone concentrations (0.08 +/- 0.02 ng/ml) remained unchanged. We conclude that during pubertal maturation in primate species there is increased synthesis and release of GnRH from a population of GnRH neurons that are active prior to puberty.     

Comparison of the duration and power spectral changes of monopolar and bipolar M waves caused by alterations in muscle fibre conduction velocity

The muscle compound action potential (M wave) recorded under monopolar configuration reflects both the propagation of the action potentials along the muscle fibres and their extinction at the tendon. M waves recorded under a bipolar configuration contain less cross talk and noise than monopolar M waves, but they do not contain the entire informative content of the propagating potential. The objective of this study was to compare the effect of changes in muscle fibre conduction velocity (MFCV) on monopolar and bipolar M waves and how this effect depends on the distance between the recording electrodes and tendon. The study was based on a simulation approach and on an experimental investigation of the characteristics of surface M waves evoked in the vastus lateralis during 4-s step-wise isometric contractions in knee extension at 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90% MVC. The peak-to-peak duration (Durpp) and median frequency (Fmedian) of the M waves were calculated. For monopolar M waves, changes in Durpp and Fmedian produced by MFCV depended on the distance from the electrode to the tendon, whereas, for bipolar M waves, changes in Durpp and Fmedian were largely independent of the electrode-to-tendon distance. When the distance between the detection point and tendon lay between approximately 15 and 40 mm, changes in Durpp of bipolar M waves were more pronounced than those of distal monopolar M waves but less marked than those of proximal monopolar M waves, and the opposite occurred for Fmedian. Since, for bipolar M waves, changes in duration and power spectral features produced by alterations in MFCV are not influenced by the electrode-to-tendon distance, the bipolar electrode configuration is a preferable choice over monopolar arrangements to estimate changes in conduction velocity.     

Dynamic aspects of the LHRH system associated with ovulation in the little brown bat (Myotis lucifugus)

Adult female bats were collected from natural roosting sites in pre-ovulatory and post-ovulatory conditions. LHRH neurones of these animals were examined using light and electron microscopic immunocytochemistry, and LHRH tissue contents were measured by radioimmunoassay. Comparisons between the two groups of bats revealed that the number of LHRH perikarya detected immunocytochemically, as well as hypothalamic LHRH content, were significantly reduced in post-ovulatory animals. Distributions of immunoreactive perikarya were, however, strikingly similar in both groups. The reduction in immunoreactive cell number observed after ovulation was therefore not restricted to an anatomically defined subset of neurones, but was evident throughout the population. The projection of LHRH-immunoreactive fibres that extend into the pituitary neural lobe in this species also exhibited changes related to endocrine condition. Morphometric indices of fibre density in the neural lobe were significantly reduced in post-ovulatory bats, as was LHRH content of the lower infundibular stalk and neural lobe. Fine structural study of perikarya revealed complex anatomical interactions between LHRH-immunopositive elements, especially in post-ovulatory bats. These interactions included direct apposition of perikarya, as well as more elaborate networks involving various combinations of perikarya and large- and small-caliber processes. These changes in the LHRH system associated with ovulation suggest reduction of stored peptide within perikarya and depletion from terminals within the lower infundibular stem and neural lobe. Parallel reductions in hypothalamic and neural lobe LHRH content during the periovulatory period support the hypothesis that the neural lobe component of the system contributes to control of gonadotrophin secretion in this species. Finally, increased complexity of anatomical contact between components of the LHRH system may be related to activation of this cell population in spring.     

Comparison of NADPH diaphorase activity in the brains of hamsters infected with scrapie strains 139H or 263K or with normal hamster brain homogenate

Previous studies showed that the histopathological changes found in the brains of scrapie-infected animals included amyloid plaque formation, vacuolation, gliosis and neuronal and neurite degeneration. There were differences in the histopathological findings as a function of the scrapie strain-host combination. NADPH-diaphorase (NADPH-d) has been shown to be a selective histochemical marker for neurons containing nitric oxide (NO) synthase. Neuronal cell damage caused by NOS in brain has been reported to be associated with many neurodegenerative diseases. In this study, we used NADPH-d histostaining to investigate changes in the NOS system in brains of 139H- and 263K-infected hamsters and compared the results to normal hamster brain (NHB) injected animals. We observed that some of the NADPH-d histostaining neurons in the cortex of scrapie-infected hamsters appeared to be atrophic: the neurons were smaller and had fewer neurites. The NADPH-d histostaining intensity of neurons or astrocytes in septum, thalamus, hypothalamus and amygdala of 139H- and 263K-infected hamsters was greater than in control hamsters. Astrocytes in the thalamus, hypothalamus and lower part of the cortex (layers 4 to 6) in 263K-infected hamsters were more intensely stained for NADPH-d than in either 139H-infected hamsters or controls. Our results suggest that changes in NADPH-d system might play a role in the diversity of scrapie induced neurodegenerative changes.     

Hypothalamic LHRH and plasma LH levels after electrical stimulation of the deafferented medial basal hypothalamus

The medial basal hypothalamus (MBH) of female rats was surgically isolated on the morning of proestrus and luteinizing hormone (LH) levels in the blood were determined before and after electrochemical stimulation (ECS) of the disconnected arcuate-median eminence (ARC-ME) region either on the same afternoon (Group 1) or on the 5th postoperative day (Group 2). Animals of Groups 3 and 4 were stimulated and sampled for LH on the 5th or 10th postoperative day, respectively, these rats having been primed with 5 micrograms estradiol injected daily throughout the experimental period. ECS resulted in a significant rise of plasma LH level in Group 1 and caused full ovulation as evaluated by the presence of ova found the next morning in the Fallopian tubes. Rats of Groups 2-4 failed to show any changes in plasma LH, and no ovulation was observed following ECS. Immunohistochemical examination of the brains revealed that axons staining for the luteinizing hormone-releasing hormone (LHRH) in the ARC-ME region remained at control levels 3 days after deafferentation (e.g. Group 1), whereas a marked decrease or complete absence of these structures was observed 8 or 13 days following surgical isolation of the MBH (e.g. Groups 2-4). These studies strongly support the view that no LHRH synthesizing perikarya are located within the MBH of the rat.     

Nerve growth factor changes the relative levels of neuropeptides in developing sensory and sympathetic ganglia of the chick embryo

The effects of chronic nerve growth factor administration on the development of neuropeptides in the embryonic chick peripheral nervous system were quantitated by radioimmunoassays. Starting at embryonic Day 3.5, daily doses of 20 micrograms of nerve growth factor (NGF) increased the substance P content of lumbosacral spinal sensory ganglia at all ages studied (Days 10-14), while having no effect on substance P levels of thoracic sensory ganglia. In contrast, the contents of somatostatin were increased in both thoracic and lumbosacral ganglia, but only at comparatively late time points (Day 14). Nerve growth factor administration was also found to decrease the somatostatin contents of lumbosacral paravertebral sympathetic ganglia at early time points (Day 8) while increasing levels at later stages (Day 14), thus acting to accelerate the normally occurring developmental changes in level of this peptide. These changes were shown to be specific for somatostatin by demonstrating that NGF increased tyrosine hydroxylase levels in sympathetic neurons at Day 8, and had no effect on sympathetic vasoactive intestinal polypeptide levels at Day 14. It has been concluded that exogenous NGF does not simply act to increase or prolong the expression of neuron-specific phenotypes in the chick, but rather its action is time and location dependent to accelerate development.     

Steroid imprinting and modulation of sexual dimorphism in the luteinizing hormone-releasing hormone neuronal system

Summary 1. Sex differences in the control of gonadotropin secretion and reproductive functions are a distinct characteristic in all mammalian species, including humans. Ovulation and cyclicity are among the most distinct neuroendocrine markers of female brain differentiation, along with sex behavioral traits that are also evident in different species.2. The luteinizing hormone-releasing hormone (LHRH) neuronal system is the prime regulator of neuroendocrine events leading to ovulation and hormonal changes during the menstrual cycle and, as such, is the potential site where many of these sex differences may be expressed or, at the very least, integrated. However, until recently, no significant differences were seen in LHRH neurons between male and female brains, including cell number, pattern of distribution, and expression of message or peptide (LHRH) levels.3. Recently, we reported that galanin (GAL), a brain-gut peptide, is coexpressed in LHRH neurons and that this coexpression is sexually dimorphic. When GAL is used as a marker for this neuronal system, it is clear that estradiol as well as progesterone profoundly affects the message and expression of the peptide and that this regulation, at least in rodents, is neonatally predetermined by gonadal steroid imprinting.4. Changes in GAL expression and message can also be seen at puberty, during pregnancy and lactation, and in aging, all situations that affect the function of the LHRH neuronal system. Using an immortalized LHRH neuronal cell line (GT1) we have recently observed that these neurons express estrogen receptor (ER) and GAL and that estradiol can increase the expression of GAL, indicating functional activation of the endogenous ER.     

Effects of ethanol on rat hypothalamic luteinizing hormone releasing hormone. A study utilizing radioimmunoassay

To further understand the mechanism of action by which ethanol (ETOH) decreases plasma luteinizing hormone (LH) levels, the effects of multiple i.p. injections of EOH (1.0--1.5 g/kg) or saline on hypothalamic luteinizing hormone releasing hormone (LHRH) and plasma LH concentrations were evaluated in intact and castrate male rats. After injections, animals were decapitated, brains rapidly removed, and blocks containing the hypothalamus [with median eminence (ME)] were isolated. Hypothalami were subjected to acetic acid extraction and LHRH content quantitated via radioimmunoassay (RIA). Hypothalamic LHRH was found to be inversely correlated with plasma LH. In response to castration, both saline and ETOH-treated rats showed a decrease in hypothalamic LHRH content with a concomitant increase in plasma LH; however, the ETOH-treated animals retained significantly greater concentrations of LHRH and showed significantly lower plasma LH levels when compared to saline-treated controls. Likewise, ETOH-treated intact animals showed significant increases in LHRH content, with LH levels remaining significantly lower than the saline-treated intact controls. Thus, these data from both intact and castrate rats provide evidence to support the hypothesis that alcohol-induced decreases in LH levels are due to a diminished release rate of hypothalamic LHRH.     

Localization of avian LHRH-immunoreactive neurons in the hypothalamus of the domestic fowl,Gallus domesticus,and the Japanese quail,Coturnix coturnix

The localization of LHRH-containing perikarya and nerve fibers in the hypothalami of the domestic fowl and Japanese quail was investigated by means of the specific immunoperoxidase ABC method, using antisera against chicken LHRH-I ([Gln8]-LHRH), chicken GnRH-II ([His5-Trp7-Tyr8]-LHRH[2-10]) and mammalian LHRH ([Arg8]-LHRH). Chicken LHRH-I-immunoreactive perikarya were sparsely scattered in the nucleus preopticus periventricularis (POP), nucleus filiformis (FIL) and nucleus septalis medialis (SM), and in bilateral bands extending from these nuclei into the septal area in both species. A few reactive perikarya were also observed in the nucleus accumbens (Ac) and lobus parolfactorius (LPO). Numerous cLHRH-I-immunoreactive fibers were widely scattered in the preoptic, septal and tuberal areas, and were densely concentrated in the external layer of the median eminence and in organum vasculosum of the lamina terminalis (OVLT) in both species. Anti-mammalian LHRH serum cross-reacted weakly with perikarya and fibers immunoreactive to anti-cLHRH-I serum in normal chicken and quail. Anti-cGnRH-II[2-10] serum immunoreacted with magnocellular neurons distributed in the rostral end of the mesencephalon along the midline close to the nervus oculomotorius (N III). These perikarya were apparently different from cLHRH-I immunoreactive neurons. No immunoreactive cells and fibers against anti-cGnRH-II[2-10] were observed in the hypothalamus and median eminence of the chicken or quail. Anti-cGnRH-II[2-10] bound specifically with cGnRH-II.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of age and biotin status on postnatal development of plasma biotinidase activity in rats

Biotinidase activity was measured in plasmas of 1-, 7-, 14-, and 21-day-old rats from control dams and dams that had been fed a biotin-depleting diet from Day 15 of gestation. Biotinidase activity increased significantly in the plasma of rats from control and depleted mothers until Postnatal Day 14, after which there was a small but significant decline at Day 21. Differences between the mean activities of the two groups of pups on each sampling day were not significant and there were no significant differences in activity levels attributable to sex. Plasma albumin concentrations increased from birth until Day 21, and plasma biotinidase activity and albumin concentration were significantly correlated (r = +/- 0.43). We suggested that these two proteins may be controlled by a common mechanism in the early postnatal period, and that biotin deficiency does not affect the development of biotinidase activity. Because biotin-depleted neonatal pups show developmental changes in biotinidase activity similar to those of human newborns, and they can be produced reliably by depleting dams from Day 15 of gestation, they may be useful models for studying the developmental abnormalities associated with human biotinidase deficiency.     

Immunocytochemical localization of luteinizing hormone-releasing hormone within the olfactory bulb of pigs

LHRH was immunocytochemically localized within the olfactory bulb of prepubertal (n = 3), ovariectomized (n = 3), and hypophyseal-stalk-transected (HST) female pigs (n = 3). Perikarya of LHRH-immunoreactive neurons of all pigs were sparsely distributed mostly in the rostral half of the olfactory bulb, along the ventromedial and ventrolateral edge of the olfactory nerve layer, or at its interace with the glomerular layer. Processes from these cells and other LHRH containing axons either entered individual glomeruli forming a network within its interior or coursed around glomeruli penetrating into the external granular layers. Additional fibers penetrated into similar regions of the accessory olfactory bulb. Irregularly shaped perikarya were also detected within the internal granular layer of the ventral olfactory bulb, but only in tissue from HST pigs. From analysis of serial sections, there was no evidence of LHRH projections across the olfactory peduncle that connects the olfactory bulb with adjacent brain regions. If olfactory LHRH neurons are involved in reproductive behavior and physiology in the pig, this pathway involves additional unidentified intervening neurons. Endocrine factors probably influence the expression of immunoreactive LHRH in the internal granule layer, since their presence was revealed only in HST pigs.