Perivascular regions of the rat neural lobe

Summary Neural lobes and portions of occipital cortex from rats were examined electron microscopically following fixation in 4% tannic acid in 2.5% glutaraldehyde. The procedure allowed a clear demonstration of the perivascular space and intercellular spaces in both tissues. The perivascular spaces in the neurosecretory tissue was far more extensive than in the neural tissue and the role of this region in relation to the process of neurosecretion is discussed.     

Effects of hormone-releasing stimuli on the area of the perivascular space in the neural lobe of the rat

Summary Neural lobes from rats subjected to neurohypophysial hormone-releasing stimuli were examined electron microscopically following fixation in 4 % tannic acid in 2.5 % glutaraldehyde. This fixation allowed the delineation of the perivascular space in the neural lobe tissue. Measurement of the area of the perivascular space showed that it was significantly increased in the rats subjected to vagal stimulation and intraarterial calcium ions compared to the control rats. The rats which had been subjected to haemorrhage as a hormonereleasing stimulus did not show any significant change in the area of the perivascular space. The significance of these findings in relation to hormone release is discussed.     

Upregulation of the p75 low-affinity neurotrophin receptor by phagocytically active perivascular cells in the rat neural lobe

Expression of the p75 low-affinity neurotrophin receptor (p75NTR) was investigated immunocytochemically at the light and ultrastructural level during the axonal degeneration that follows partial denervation of the rat neural lobe (NL) and following systemic administration of lipopolysaccharide (LPS). A significant increase in the intensity and extent of p75NTR immunoreactivity in the NL of partially denervated animals compared with age-matched, sham-operated controls was observed at 5-10 days postdenervation, with immunoreactivity returning to control values by 35 days. Dual-label confocal comparison of p75NTR localization with that of the C3bi complement receptor, a microglial marker, and S100, an astrocyte-specific Ca2+-binding protein, revealed no colocalization. Immunoelectron-microscopic examination demonstrated that the p75NTR immunoreactivity is present in a subpopulation of cells located within the extensive perivascular space of the NL. No examples of p75NTR-immunoreactive pituicytes or endothelia were observed at the light or ultrastructural level. Dense p75NTR immunoreactivity was frequently observed surrounding endocytotic omega profiles of plasmalemma engulfing extracellular debris as well as lining vacuoles within the cytoplasm of perivascular cells. The association of p75NTR with phagocytosis was confirmed by confocal microscopy, showing the presence of p75NTR in all cells expressing the ED-1 antigen, which is restricted to the lysosomal membrane of phagocytes (Damoiseaux et al. 1994). Likewise, a marked increase in p75NTR and ED-1 immunoreactivity was observed in the NL following systemic administration of LPS. These results suggest a strong correlation between modulation of p75NTR immunoreactivity and conditions that induce high levels of phagocytic activity by perivascular cells in the NL of the rat. Implications for understanding the mechanisms by which phagocytes may support compensatory responses to neuronal injury are discussed.     

On the fine structure of the perivascular cells in the neural lobe of rats

Summary The purpose of this study is to investigate the ultrastructural features of perivascular cells as found in the neural lobe of the hypophysis. In particular, an inquiry was made into the nature of the relationship of such cells to neurosecretory fibers and endings. The latter, in fact, are often invaginated within the perivascular cells and enveloped by their processes; furthermore, they often reveal a certain number of empty granules as well as characteristics of degenerative nature. In the course of this study the localization of the perivascular cells has been investigated as well as that of their processes within the extensive interlobular network typical of the hypophysial neural lobe of rodents. Based on the data gathered, the hypothesis is put forward that the perivascular cells play an important role in the turnover of neurosecretory endings, both under physiological and experimental conditions, contributing thereby to the release of post-hypophysial hormones.With the technical assistance of Vincenzo Panetta.     

Endogenous opioids inhibit the in vitro release of endogenous dopamine preferentially in the neural lobe of the rat neurointermediate lobe

The release of endogenous dopamine (DA) from the in vitro incubated combined neurointermediate lobe (NIL) or isolated neural lobe (NL) was studied. In the presence of the DA uptake inhibitor GBR 12921 (200 nM), electrical stimulation of the pituitary stalk caused an increase of the outflow of DA from the NIL in a frequency-dependent manner. Naloxone (1 microM) enhanced the DA release from the NIL evoked by electrical stimulation at 7 or 15 Hz by about 40%, but had no effect on DA release evoked by stimulation at 3 Hz. When the electrical stimulation was carried out at 15 Hz, the evoked DA release (expressed as fraction of the DA tissue content) from the NL amounted to only 15% of that from the combined NIL. Naloxone (1 microM) increased the evoked DA release from the isolated NL by 242%. Thus, the effect of naloxone on DA release from the combined NIL may be confined mainly to the NL. In conclusion, DA release from the NL is under inhibitory control of endogenous opioids released from the NL during stimulation at 7 or 15 Hz. Beta-Endorphin, known to be released spontaneously at a high rate from in vitro incubated NILs, appears to lack inhibitory effects on DA release from the NIL.     

Basal gonadotropin hormone release rates during the period of selective follicle-stimulating hormone release in the juvenile female rat

There are situations in which adult female rats release increased amounts of follicle-stimulating hormone (FSH) independent of increased luteinizing hormone (LH) release. This results from, at least in part, a selective increase in the basal FSH release rate. We investigated whether an increase in the basal FSH release rate is contributory to the rise in serum FSH levels which occurs independent of a rise in serum LH levels in the immature female rat. Rats had high serum FSH concentrations on days 7 and 15 after birth, low serum FSH levels on day 23, and low serum LH levels on all three days. In contrast, anterior pituitary gland (APG) FSH and LH concentrations and contents increased from day 7 to day 15 and the contents increased further from day 15 to day 23. Similarly, basal FSH and LH release rates per mg APG or per APG, as assessed by measurement of FSH and LH released into culture medium containing APG(s) from different aged rats, increased from day 7 to day 15 but did not increase further between days 15 and 23. The results indicate that unlike situations observed to date in adult female rats, a mechanism(s) other than an increase in the basal FSH release rate is involved in selective FSH release in the immature female rat.     

Rat posterior lobe modulates growth hormone release in vivo and in vitro

Electrical stimulation of hypophysical posterior lobes $\text{in vivo}$ evokes a significant decrease of plasma growth hormones (GH) and an increase of plasma corticotropin (ACTH) concentrations. Electrical stimulation of posterior lobes $\text{in vitro}$ evokes the simultaneous release of GH inhibiting factor(s) (GHRIF) and ACTH releasing factor(s) (CRF) into the medium. Pretreatment of media with thioglycolate abolishes the CRF and GHRIF activity, but reveals GH releasing factor(s) (GHRF). Median eminence extracts and vasopressin have potent GHRF and CRF activity. Vasopressin may account fully for the CRF and partially for GHRF activity. Results suggest that hypothalamo-neurohypophysical axons release GHRIF, vasopressin and possibly a GHRF into a portal circulation to modulate the secretion of GH and ACTH.     

Reserpine inhibits release of vasopressin from the neural lobe of the pituitary in dehydrated rats

Summary The effects of reserpine on the osmotically induced release of pituitary vasopressin were studied (i) by measuring the urinary excretion and the vasopressin content of the neural lobe of the pituitary, and (ii) by examining the ultrastructural morphology of axons in the neural lobe of dehydrated rats. After water deprivation for two days, control rats displayed characteristic antidiuretic response including a 75% reduction of urinary excretion and a sixfold decrease in vasopressin content of the neural lobe associated with a dramatic depletion of neurosecretory granules in corresponding axons. In contrast, when they received two daily injections of reserpine, animals dehydrated for two days showed both urinary excretion and vasopressin contents in the neural lobe that remained at levels comparable to those measured in the normally hydrated rats. Additionally, neural-lobe axons of such dehydrated, reserpine-treated rats displayed a normal amount of neurosecretory granules. These data indicate that reserpine inhibits release of vasopressin from the neural lobe and favour the concept of a facilitatory role of the catecholaminergic innervation in the control of hypothalamo-neurohypophysial vasopressin-secreting neurons.     

In vitro and in vivo iodination of human thyroglobulin in relation to hormone release

Reduced and S-alkylated thyroglobulin (Tgb) from different species were shown by SDS-PAGE to contain small peptides (from 45-9 kDa) rich in thyroxine. Several hypotheses were proposed to explain their origin. The polypeptide composition of iodine-poor (Tgb A) and normally iodinated (Tgb B) human Tgb prepared by two different procedures (one minimizing and the other favoring post-mortem proteolysis) was compared in the native state and after in vitro iodination. Results show that one of the hormonogenic sites of human Tgb is part of a domain of the molecule most susceptible to proteolysis, especially when it is very iodinated.     

Canada goose posterior lobe pituicytes: seasonal ultrastructural changes in relation to axonal release of neurosecretion

Changes in ultrastructure of the posterior lobe pituicytes during six phases (Spring premigratory, Spring postmigratory, breeding, moulting, Fall premigratory and Fall postmigratory) of the annual life cycle of the migratory Canada goose, were studied. Pituicyte nuclear volume in females was significantly greater than that in males during the Spring premigratory and moulting phases. In the Fall postmigratory phase, nuclear volume in males was greater than that in females. During Spring premigratory phase, the axonal endings in males contained fewer neurosecretory granules (NSG) than those in the Fall postmigratory phase. In females, however, the number of NSG was relatively more than that in males in the Spring premigratory phase but fewer in the moulting phase. Nuclear volume is correlated with pituicyte function in releasing NSG containing arginine vasotocin (AVT) from axonal endings. The significance of AVT release is discussed in relation to initiation of reproductive and migratory behaviour, and to osmotic and metabolic regulation.     

Gonadotrophic cells of the rat hypophysis and their relation to hormone production

Summary The results of these experiments clearly indicate that the PAS-red and PAS-purple gonadotrophs of the rat anterior hypophysis are functionally as well as tinctorially distinct cell types. The PAS-red cells located peripherally in control animals (peripheral gonadotrophs of Purves and Griesbach) produce LH. Following castration they begin to appear in greater numbers in the central areas and by 45 days following castration they are predominant in both central and peripheral portions of the glands. At this time after castration the gonadotrophic content of the pituitary gland is predominantly luteinizing in character.The PAS-purple cells which are found in the central portions of control glands (central gonadotrophs of Purves and Griesbach) produce FSH. They appear peripherally following castration and are the predominant type of gonadotrophic cell in the glands of short term (10-day) castrates. The gonadotrophic content of such glands is chiefly FSH as shown by bioassay.By 45 days after castration the LH producing peripheral gonadotrophs and the FSH producing central gonadotrophs have lost their characteristic distribution patterns. Obviously, then, these two gonadotrophic types cannot be accurately followed on the basis of a restricted regional location in the pituitary but must be differentiated on the basis of specific cytological features and staining characteristics.This study was supported by USPHS Grant RG 4723 and by contract between Office of Naval Research, Department of the Navy, and University of Texas.     

Morphology of the neural lobe of the hypophysis in rats treated with furosemide. II. Pituicytes

Following three days of furosemide administration, the characteristics of the pituicytes in the neural lobe of the rat hypophysis were studied. The pituicytes of the animals which were treated with furosemide and had access to water during the treatment showed scarce cytoplasm organelles, conferring them with a clear appearance (light pituicytes), and a frequent finding was neurosecretory axons enclosed in their own cytoplasm. In the animals deprived of water during treatment, the pituicytes were dense (dark pituicytes) elongated, with a well-developed Golgi apparatus, numerous profiles of smooth endoplasmic reticulum and frequent annular gap junctions. It is concluded that the dehydration induced by the diuretic agent is the cause of the prevalence of the dark pituicytes.     

Stress-induced ACTH release after removal of the hypothalamus in rats with atrophied neural lobe.

Twenty-four hours after isolation of the pituitary by surgical removal of the medial hypothalamus, i.e. in rats with pituitary island, E. coli endotoxin significantly increased the plasma corticosterone level. Atrophy of the neural lobe, due to pituitary stalk section performed one month prior to removal of the medial hypothalamus, did not prevent the increase of ACTH release by E. coli endotoxin. E. coli endotoxin-induced ACTH release in MBH-deprived animals does not appear to be a function of mechanisms operating only in the innervated lobe.     

Morphology of the neural lobe of the hypophysis in rats treated with furosemide. I. Neurosecretory axons

The changes found in the neurosecretory axons of the neural lobe of the hypophysis were studied in rats treated with furosemide for three days: over the whole period of treatment, the animals, according to each group, were deprived or not of water. In the animals with free access to water the axons contained neurosecretory granules with scarce content and low electron density; only some of them had vacuoles and autophagic bodies. In the animals deprived of water axon morphology was variable and axons showing vacuoles and autophagic bodies were abundant, as well as those presenting lamellar and dense bodies and also those in which filaments prevailed. The most outstanding modifications in this last group of animals were related to the dehydration and were not found in those animals which were given access to water again and which were in consequence rehydrated.     

Ultrastructure of the rat neural lobe during recovery from hypertonic saline treatment

Summary Neural lobes from rats which had been allowed to drink hypertonic saline for five days were examined electron microscopically and by bioassay of oxytocin levels. The profound changes in the ultrastructure were examined and the reversal of these changes in morphology was followed until the hormone levels returned to normal. The recovery of the gland as measured by the two parameters of structure and hormone content indicated that the morphological recovery apparently preceded the hormone level recovery, a factor which might be explained by continued release of hormone during the period of recovery.     

Effect of phenoxybenzamine on luteinizing hormone release in the female rat

To clarify the mode of action of phenoxybenzamine, an alpha adrenergic blocking agent, its effects upon plasma LH levels in ovariectomized rats and upon the ovulatory LH surge expected between 1400 and 1600, the critical period, on the day of proestrus in normal rats were studied. A single injection of phenoxybenzamine, 20 mg/kg, given at 1300 on the day of proestrus bokced ovulation (1 out of 7 ovulating), while plasma LH did not differ from controls between 1500 and 1600. An additional injection of 20 iu HCG at 1500 prevented the ovulation block (83% ovulating). A single phenoxybenzamine injection at 1700 failed to prevent ovulation (5 out of 7 ovulating). The beta adrenergic blocking agents, propanolol and MJ 1999, did not affect ovulation. Treatment with phenoxybenzamine for 2 days, 20mg/kg/day, for 8 days, 10mg/kg/day, were did not prevent the rise causing a reduction in blood flow through the ovary rather than acting as a neurogenic stimulus in the hypothalamus.