Biomedical subjects

G T Campbell

Publications and source records attributed to G T Campbell.

At least 19 recordsLinked to original sources

PubMed1999

Interactions between estrogen, tamoxifen, octylphenol, and two polychlorinated biphenyls in murine splenocytes.

Prior exposure of cultured murine splenocytes to 17beta-estradiol (E) protects them from the membrane disrupting effects of the xenoestrogen 4-tert-octylphenol (OP). Using splenocytes isolated from male Balb/c mice, we tested whether (a) the xenoestrogen, 2', 3', 4', 5'-tetrachloro-4-biphenylol (PCB-OH), or the polychlorinated biphenyl, 3, 3', 4, 4'-tetrachlorobiphenyl (PCB 77), which displays both estrogenic and anti-estrogenic actions, would compromise the membrane integrity of the cells and (b) E or tamoxifen (TX), another ligand for the E receptor, would protect the membranes of cells exposed to the agents. We also examined possible interactions between OP, PCB-OH, and PCB 77 on the cells. Splenocytes were cultured for 24 hr. Concentrations of OP (10(-5)-10(-9) M), PCB-OH (10(-6)-10(-16) M), or PCB 77 (10(-8)-10(-12) M) significantly compromised the membrane integrity of the cultured splenocytes in a dose response manner. Concentrations of E as high as 10(-5) M or TX as high as 10(-7) M were without effect. Incubation of splenocytes in medium containing E or TX at 10(-7) M for 2 hr prior to the subsequent addition of either OP, PCB-OH or PCB 77 (final concentrations of 10(-7), 10(-7), or 10(-8) M, respectively) blocked the membrane disrupting effects. Incubation of splenocytes in medium containing 10(-7) M E starting 2 hr after the addition of OP or PCB 77 or incubation of splenocytes in medium containing 10(-7) M TX starting 2 hr after the addition of OP or PCB-OH did not block the damaging effects of OP, PCB 77, or PCB-OH on the cell membranes. No interactions were observed when various combinations of OP, PCB-OH, or PCB 77 were used. These data suggest that: (a) TX acts like E in this system, (b) a prior response of splenocytes to E or TX can protect them from the potential cytotoxic effects of OP, PCB-OH, or PCB 77; and, (c) OP, PCB-OH, and PCB 77 were not additive in their actions.

Animals↗

PubMed1997

Estrogen can protect splenocytes from the toxic effects of the environmental pollutant 4-tert-octylphenol.

Four-tert-octylphenol (OP), an environmental pollutant, exerts apoptotic effects on cultured mouse splenocytes. Although OP binds to estrogen receptors, these apoptotic effects are not exerted by 17 beta-estradiol (E). It remained possible that OP might bind to estrogen receptors and subsequently exert apoptotic effects not exerted by E after it binds to the same receptors. It also remained possible that E-primed splenocytes might respond to OP differently than splenocytes not exposed to E. Thus, we investigated OP and E interactions on the viability of mouse splenocytes in culture. The total number of splenocytes (cells stained and not stained with trypan blue) was not altered or altered slightly after incubation with any agent for 24 h. Incubation of splenocytes in medium containing 5 x 10(-5) or 5 x 10(-7) M OP decreased the percentage of viable cells by only approx 47% and 25%, respectively. The addition of 0.8 x 10(-5) to 0.8 x 10(-9) M E to cultures was without effect or decreased the percentage of viable cells by only approx 5%. The addition of these concentrations of E simultaneously with or at 2 h after the addition of 5 x 10(-5) M or 5 x 10(-7) M OP to cultures did not interfere with the OP-induced decreases in cell viability. By contrast, incubation of splenocytes in medium containing E for 2 h prior to the subsequent addition of either dose of OP blocked the OP-induced decreases in cell viability in a dose-response manner. There was a marked reduction in the percentage of viable cells (70%) when splenocytes were incubated with 0.5 x 10(-5) M dexamethasone. The addition of 0.8 x 10(-5) M E at 2 h prior to the addition of dexamethasone did not prevent the decreased cell viability. Incubation of cells in medium with 0.8 x 10(-5) M testosterone caused a small decrease in splenocyte viability similar to that observed with E. However, unlike E, the addition of testosterone at 2 h prior to the addition of 5 x 10(-5) M OP did not prevent the OP-induced decrease in cell viability. These data suggest the presence of estrogen receptors in some splenocytes. They also suggest that if OP binds to these estrogen receptors or other receptors in the absence or initial presence of E, the resulting effect is toxic to the cells. By contrast, exposure of splenocytes to E prior to their exposure to OP can prevent the toxicity of OP.

Animals↗

PubMed1996

Toxic effects of octylphenol on cultured rat and murine splenocytes.

Alkylphenol polyethoxylates and alkylphenols, such as 4-tertoctylphenol (OP), are environmental contaminants. Because these compounds are toxic to aquatic animals, we studied the effects of OP on splenocytes removed from male Fischer 344 rats or male Balb/c mice and cultured in vitro. Cell viability was assessed by trypan blue exclusion after 5 or 27 hr of culture. Culture with 0.08% ETOH (vehicle) or any dose of OP did not alter total cell number or the percentage of viable cells after 5 hr. Culture of cells with two different alkylphenol polyethoxylates for 5 hr resulted in the loss of all cells. The percentages of viable rat or mouse cells after 27 hr of culture were decreased significantly by 10(-12) M OP or greater concentrations. The actions of OP, dexamethasone (DEX), and 17 beta-estradiol on rat splenocytes were compared. Dexamethasone was more toxic than OP after 24 hr of culture; 17 beta-estradiol was not toxic. Dexamethasone and OP, but not 17 beta-estradiol, caused significant nuclear condensation after 3 hr of culture (acridine orange staining) or 4 hr of culture (propidium iodide staining). The toxicity of 10(-6) M OP, but not that of 10(-6) M DEX, was eliminated when mouse splenocytes were cultured in Ca2+ -free medium. Significantly more mouse splenocytes containing free 3'-OH DNA ends were detected by activated cell sorter analyses when the cells had been incubated for 4 hr with 10(-4) or 10(-6) M OP or 10(-6) M DEX. The results of these studies demonstrate that OP is toxic to cultured rat and mouse splenocytes and suggest that this toxic effect is exerted, at least partially, through Ca2+-dependent apoptosis.

Animals↗

PubMed1996

Changes in percentages of adenohypophysial gonadotrophs associated with the sex-specific, selective increase in serum follicle-stimulating hormone concentration in the juvenile female hamster.

In normal hamsters, we investigated whether the sex-specific, selective increase in serum FSH concentration in the juvenile female was associated with sex-specific changes in the percentages of adenohypophysial gonadotrophs. Serum LH concentrations did not rise between Day 4 and Day 19 in either sex and did not differ significantly between the sexes on Days 4, 7, 12, 14, and 19 after birth. Serum FSH concentrations were about 2-fold higher on Days 7, 12, and 14 than on Days 4 or 19 in males. In females, serum FSH rose markedly between Days 4 and 7, declined slightly by Day 12, rose to peak levels by Day 14, and declined slightly by Day 19 to levels not different from those seen on Day 7. Body weights rose between Days 4 and 19 and were similar in both sexes. There were no sex differences in pituitary gland weights, which rose between Days 4 and 12 and did not increase significantly further by Day 19. On Day 0, the percentages of immunoreactive LH and FSH cells were about 6 and 1%, respectively, in both sexes. These percentages increased progressively between Days 0 and 7 and between Days 7 and 14. On Day 7, but not on Day 14, the percentages of LH and FSH cells were greater in females than in males. There were more LH than FSH cells in males on Days 0, 7 and 14, and in females on Day 0 but not on Day 7 or 14. Matching of 10 FSH cells per gland with LH cells in serial sections of each of 30 glands showed FSH immunoreactivity to occur only in cells staining for LH. In hypophysectomized-gonadectomized adult hamster hosts with allografts of neonatal pituitary glands beneath the renal capsule, we investigated whether these sex-specific changes in the percentage of cells might be predetermined by the time of birth or dependent on sex differences in the internal environment existing in the postnatal hamster. Groups consisted of male donors-male hosts, male donors-female hosts, female donors-female hosts, and female donors-male hosts. The percentages of LH cells in allografts in all four groups increased from Days 0 to 7 and from Days 7 to 14. Percentages of LH cells on Day 14 in all four groups were not different from those in age-matched male or female adenohypophyses in situ. In contrast, the mean percentages of FSH cells were low (about 1-3%) on Days 0, 7, and 14 in all four groups. In other males hosts, administration of a low dose of LHRH for 7 days did not alter the percentage of LH cells in male allografts but increased the percentage of FSH cells to approach that observed in age-matched male adenohypophyses in situ. Administration of a larger dose of LHRH for 7 days to other male hosts with male allografts increased the percentages of LH and FSH cells to percentages not different from those in age-matched female adenohypophyses in situ. Matching of 10 FSH cells/allograft with LH cells in serial sections of each of 58 allografts showed FSH immunoreactivity to occur only in cells staining for LH. The results of experiments conducted on normal hamsters demonstrate that more marked increases in the percentages of adenohypophysial LH cells and FSH cells occur in females than in males in association with the onset of the selective increase in serum FSH levels in females. The results of experiments employing allografts suggest that the greater increase in LH and FSH cells in females is due to sex differences in the internal environment existing in the postnatal hamster, which can be accounted for by differences in LHRH secretion, rather than to inherent differences between female and male adenohypophyses at the time of birth. We conclude that the greater increases in gonadotrophs observed in female hamster pups on Day 7 after birth and the accompanying sex-specific, selective elevation in serum FSH concentration are probably due to sex differences in LHRH secretion during the juvenile period.

Animals↗

PubMed1996

Adenohypophysial allografts releasing prolactin decrease prolactin mRNA concentration in the host hamster's adenohypophysis in situ.

The inhibitory effects of pituitary allografts on the prolactin (PRL)-secretory system are presumed to be consequences of the unabated release of PRL by the allografts. In the present studies we used pituitary allografts in the Golden Syrian hamster to address the following questions: (a) Do allografts of adult adenohypophysial tissue which elevate serum PRL levels decrease the concentration of PRL mRNA in the host's adenohypophysis? (b) Is this effect shared by allografts of neonatal hypophysial tissue or neonatal muscle tissue which do not elevate serum PRL levels? (c) Do any of these types of allograft alter growth hormone mRNA in the host's adenohypophysis? Prolactin mRNA concentration, but not growth hormone mRNA concentration, was decreased in the adenohypophyses in situ in the hosts bearing adult adenohypophysial allografts in which serum PRL levels were elevated. In contrast, serum PRL in hosts with neonatal hypophysial or muscle allografts were not elevated and PRL mRNA levels in the adenohypophysis in situ were not decreased when compared to the levels measured in hamsters with sham transplants. Prolactin mRNA levels in hosts with neonatal muscle allografts were not different from levels in hosts with neonatal hypophysial allografts but were increased when compared to the levels measured in hamsters with sham transplants. There were no differences in PRL concentration in the adenohypophyses in situ between any of the groups. Also, PRL concentrations in neonatal hypophysial allografts were similar to those in adult adenohypophysial allografts. To our knowledge these observations are the first demonstrating that short-loop feed-back of PRL includes a decrease in PRL mRNA concentration. The observations also support the working hypothesis that PRL and not another pituitary factor exerts the negative feedback.

Animals↗

PubMed1995

Induction of cellular follicle-stimulating hormone in the hamster adenohypophysis requires intermittent stimulation by luteinizing hormone releasing hormone.

We investigated the effectiveness of continuous vs intermittent LHRH stimulation of the neonatal female anterior pituitary gland on inducing cellular FSH immunoreactivity in the Golden Syrian hamster. Neonatal female pituitary glands were grafted beneath the right renal capsules of hypophysectomized-ovariectomized adult hosts with a catheter implanted in the external jugular vein. In experiment 1, vehicle or LHRH (6 ng/h) was infused continuously or LHRH was pulsed at 1 h (6 ng) or 12 h (72 ng) intervals through the catheters for 8 days. Hamsters were decapitated for collection of trunk blood shortly after the end of treatment, and grafts were prepared for immunocytochemical staining for LH and FSH. Anterior pituitary glands removed from neonatal (day 1) and day 9 female pups also were stained for LH and FSH. The mean percentage of adenohypophysial cells staining for LH increased from 11% in neonatal pups to mean percentages (24-28%) that were similar in day 9 pups and in all groups with grafts. The mean percentage of adenohypophysial cells staining for FSH increased from 1% in neonatal pups to percentages (16-21%) that were similar in day 9 pups and in grafts in hosts administered 6 or 72 ng LHRH pulses. By contrast, the mean percentage of FSH cells did not increase in grafts in hosts administered vehicle or LHRH by continuous infusion. Serum LH concentration was low in hosts given vehicle or LHRH by continuous infusion but elevated in hosts given 72 ng LHRH pulses and in all but one host given 6 ng LHRH pulses.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Neuropeptide Y and luteinizing hormone releasing hormone synergize to stimulate the development of cellular follicle-stimulating hormone in the hamster adenohypophysis.

Luteinizing hormone releasing hormone (LHRH) stimulates the development of cellular FSH immunoreactivity in the perinatal hamster adenohypophysis. Because neuropeptide Y (NPY) can act directly on rat adenohypophysial cells to stimulate FSH and LH release and potentiate the stimulatory effect of LHRH on FSH and LH release, we investigated the effects of NPY alone and in combination with a low, ineffective dose of LHRH on inducing cellular FSH immunoreactivity in the neonatal hamster adenohypophysis. Neonatal female pituitary glands were grafted beneath the right renal capsules of hypophysectomized-ovariectomized adult hamster hosts with a catheter implanted in the external jugular vein. After treatment, hosts were decapitated and graft tissue was stained for FSH and LH immunoreactivity. The mean percentage of adenohypophysial cells that stained for FSH was low (2.8%) in grafts in hosts infused continuously with heparinized saline vehicle for 7 days. In other hosts, peptides were pulsed through the catheter every 12 h for 7 days. The mean percentage of FSH cells also was low after pulsing 6 ng LHRH or 2 micrograms NPY but increased substantially when the two peptides were pulsed simultaneously. No differences in the mean percentage of LH cells existed between any of the groups. The results demonstrate that NPY and LHRH can synergize to induce cellular FSH immunoreactivity in the neonatal female hamster.

Animals↗

PubMed1993

Effects of injection of anti-luteinizing hormone (LH)-releasing hormone serum and anti-gonadotropin-releasing hormone-associated peptide serum into neonatal rats on LH and follicle-stimulating hormone cells.

Relatively little is known regarding the potential importance of LHRH and of gonadotropin-releasing hormone-associated peptide (GAP) on the postnatal development of gonadotrophs. We investigated the effects of administration of anti (A)-LHRH serum or A-GAP serum to neonatal rats on the development of LH and FSH immunoreactivity in anterior pituitary gland (APG) cells. Serum (sheep non-immune [NSS], sheep A-LHRH, sheep A-LHRH/GAP [which bound LHRH and GAP], rabbit non-immune [NRS], or rabbit A-GAP) was injected s.c. into neonatal female and male rats on Days 1 and 3 or Days 1, 3, 5, and 7 after birth. Pups were killed on Day 5 or 9, two days after the last injection. The percentages of APG cells immunoreactive for LH or FSH increased from Day 1 to Day 5 and did not change between Days 5 and 9 in female pups treated with NSS or NRS. There was a trend for the percentages of LH and FSH cells to increase from Day 1 to Days 5 and 9 in male pups treated with NSS or NRS, but the increases were not statistically significant. In both females and males, treatment with antisera that recognized LHRH reduced the percentage of FSH cells on Day 5 and the percentages of LH and FSH cells on Day 9. Treatment with A-GAP was without effect in both sexes. There were similar percentages of LH and FSH cells in females and a lower percentage of FSH than of LH cells in males in the Day 5 and 9 controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Estrogen alters the effects of neuropeptide-Y on luteinizing hormone and follicle-stimulating hormone release in female rats at the level of the anterior pituitary gland.

In recent years, several studies have shown that neuropeptide-Y (NPY) is involved in the control of LH secretion. We determined the effects of estrogen on NPY-induced LH and FSH release in the absence or presence of LH-releasing hormone (LHRH) at the level of the anterior pituitary gland (APG). Adult female rats were ovariectomized. Fifteen to 20 days later, they were given a blank or estrogen-filled capsule subdermally and killed 17-19 h later. APG cells were isolated and cultured for 3 days in medium containing 12.5% rat serum collected at death from the same rats used to make the respective APG cell pools. The cells were then challenged for 3 h with vehicle, NPY (10(-12)-10(-6) M), LHRH (10(-9)-10(-6) M), or combinations of NPY (10(-9)-10(-7) M) and LHRH (10(-9) M). LHRH stimulated LH and FSH release from nonestrogen and estrogen-primed cells. NPY at 6.7 x 10(-8)-10(-6) M increased (P < 0.05) LH release and at 10(-6) M increased (P < 0.05) FSH release from estrogen-primed cells, but was without effect on nonestrogen-primed cells. In contrast, NPY at 10(-9)-10(-7) M potentiated the action of LHRH (10(-9) M) to increase the release of LH and FSH from nonestrogen-primed cells, but was without potentiating effects in cultures of estrogen-primed cells. The results demonstrate that 1) NPY can release LH and FSH by a direct action on estrogen-primed APG cells; and 2) NPY can potentiate the action of LHRH to increase the release of LH and FSH by a direct action on nonestrogen-primed APG cells.

Animals↗

PubMed1992

Intraperitoneal injection of chloral hydrate causes intra-abdominal adhesions and unilateral testicular atrophy in golden Syrian hamsters.

We investigated the reason for the high mortality we had observed in hypophysectomized-orchidectomized Golden Syrian hamsters that were anesthetized with intraperitoneal (i.p.) injections of chloral hydrate (CH). Intact male Golden Syrian hamsters were injected intraperitoneally with 0.1cc/100g BW of a 35% solution of CH, a 35% solution of sodium chloride, or double-distilled water. Equal numbers of hamsters in each group were injected on the right or left side of the abdomen. Within 10 days, 35% of the CH-injected hamsters were dead or had to be euthanized. Autopsy revealed severe peritonitis and adynamic ileus. CH-injected hamsters that survived gained weight at a rate similar to that of the controls. All surviving hamsters were killed 18 days after the injections. Among the surviving CH-injected hamsters, 84.6% had intra-abdominal adhesions, 61.5% had unilateral testicular atrophy, and 53.8% had a yellowish necrotic mass in the epididymal fat pad (EFP). All the lesions occurred on the side that was injected. The atrophied testes had been rendered cryptorchid due to involvement with intra-abdominal adhesions. In the water-treated controls, there were no abnormalities; whereas, in the saline controls, 75% had a mass in the EFP. Histology of the EFP mass was similar in hamsters injected with CH or hypertonic saline and suggested a diagnosis of fat necrosis. The results suggest that the mortality, the intra-abdominal adhesions, and the unilateral cryptorchidism were caused by a single i.p. injection of CH, but the fat necrosis in the EFP was probably caused by high concentrations of salt. The results further suggest that high concentrations of CH should not be injected intraperitoneally for anesthesia in chronic studies, particularly of the male reproductive system.

Adipose Tissue↗

PubMed1992

Gonadotropin-releasing hormone-induced accumulation of follicle-stimulating hormone beta-subunit messenger ribonucleic acid in adenohypophysial cells developing in an ectopic position.

We investigated the influence of LHRH on the accumulation of FSH beta messenger RNA (mRNA) in anterior pituitary glands removed from hamster pups less than 36 h old and transplanted beneath the renal capsules of adult male hamsters (hosts). Three experiments were performed in which some hosts were injected sc with LHRH (1 microgram/injection) and others were injected with vehicle. Injections were begun in the afternoon of the day of transplantation (day 1) and were given at 0800 and 1700 h for 6 days and at 0800 h on the eighth day. An additional experiment was performed in which adult male hamsters not bearing allografts were injected with the same regimen of LHRH or vehicle. The hamsters were decapitated on the eighth day of the study, 2 h after the last injection. The allografts, adenohypophyses of the hosts, adenohypophyses of hamsters without allografts, and adenohypophyses of normal adult male rats were removed and frozen on dry ice immediately. Additionally, adenohypophyses were collected from hamster pups less than 36 h old and 8 and 15 days of age. Total RNAs from some pooled specimens were electrophoresed on a formaldehyde-agarose gel. After transfer to Nytran, the RNAs were hybridized sequentially to complementary DNAs for rat FSH beta and hamster beta-actin. The rat FSH beta complementary DNA probe hybridized to a single RNA (approximately 1.7 Kb) in rat adenohypophyses. It predominantly hybridized to RNA of approximately 1.7 Kb from hamster adenohypophyses. Sometimes it hybridized to RNAs ranging in size from 0.5 Kb to 1.7 kb. The hybridization signals for all samples obtained from dot blot analyses were quantitated and normalized to the signals for beta-actin. The hybridization signals obtained from adenohypophyses of hamsters of different ages increased from 36 h of age to adulthood. The hybridization signal obtained from adenohypophyses of hamsters less than 36 h old (the same age as the donor hamsters) was similar to the hybridization signal obtained from allografts in vehicle-treated hamsters. The relative levels of FSH beta mRNA in allografts of LHRH-treated hosts were: 1) greater than the relative levels in adenohypophyses of hamsters less than 36 h old (P less than 0.05) and in allografts in vehicle-treated hamsters (P less than 0.05), 2) greater than the relative levels in adenohypophyses of 8-day-old hamsters (P less than 0.05), and 3) not different compared to the relative levels in adenohypophyses of 15-day-old hamsters and adult male hamsters.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1991

Sequence and expression of hamster prolactin and growth hormone messenger RNAs.

Complementary DNAs encompassing the complete protein-encoding regions for PRL and GH of the Syrian Golden hamster were sequenced and used as probes to examine the expression of hamster PRL and GH messenger RNA (mRNA)s. The complementary DNA (cDNA) for hamster PRL encodes a 226 amino acid preprotein which, by analogy to rat and mouse PRLs, is predicted to be processed to yield a 197 amino acid secreted protein. The hamster GH cDNA codes for a 216 amino acid preprotein predicted to yield a 190 amino acid secreted protein. Both hamster proteins are highly homologous to the corresponding rat and mouse hormones. For the secreted proteins, hamster PRL has 82% amino acid identity with rat PRL and 72% identity with mouse PRL. The rodent GH sequences are more strongly conserved, with 97-98% sequence identity between hamster, rat, and mouse GHs. The hamster hormones contain the highly conserved cysteine residues (six in hamster PRL and four in hamster GH) present in other mammalian PRLs and GHs. Neither hamster PRL nor hamster GH contains cysteine residues corresponding to the unique pair of cysteines present in hamster placental lactogen-II. The hamster PRL and GH cDNAs each hybridized to pituitary mRNAs of approximately 1 kilobase. Expression of hamster PRL and GH mRNAs was compared between 2 days of the estrous cycle (proestrus and estrus) and early, mid, and late pregnancy (days 5, 10, and 15). PRL mRNA levels in cycling hamsters were approximately 50% of those in pregnant hamsters. No other significant differences in PRL or GH mRNA levels were observed, suggesting that differences in circulating PRL and GH protein levels during the estrous cycle and pregnancy in the hamster are the result largely of factors other than changes in mRNA levels.

Amino Acid Sequence↗

PubMed1990

Luteinizing hormone (LH)-releasing hormone: effects on maintenance of immunoreactive follicle-stimulating hormone and LH in adenohypophysial cells.

We investigated the importance of LHRH on the maintenance of FSH and LH immunoreactivity in gonadotrophs. Hypophysectomized orchidectomized hamsters (hosts) each received an allograft of a 7-week-old male hamster pituitary gland beneath their right renal capsule. Starting 6 days after transplantation, hosts were injected sc, twice daily with 1 micrograms LHRH or vehicle for 16 days. Twelve hosts in each group were killed by decapitation 16 h after the last injection. Allografts from six of the hamsters in each group and pituitary glands in situ from 10-week-old normal males were prepared for histological examination. Sections of tissue were stained for FSH or LH and with hematoxylin. Allografts from the remaining hamsters were homogenized to measure FSH and LH concentrations. In allografts from the vehicle-treated hosts, 22.8% of adenohypophysial cells stained for LH, while only 16.9% stained for FSH. In allografts from LHRH-treated hosts, 22.6% and 23.8% of the adenohypophyses cells stained for LH and FSH, respectively. Adenohypophyses that developed for the same length of time in situ had 24.8% and 24.1% of the cells staining for LH and FSH, respectively. Matching of some of the FSH and LH cells in serial flip-flopped sections of tissue from all hamsters revealed that many if not all gonadotrophs contained LH. LH- and FSH-containing cells in allografts were similar in size and shape, but were smaller and more circular in profile than those observed in situ. Treatment of hosts with LHRH did not alter gonadotroph size or shape, but it did reduce allograft LH concentration and elevate the serum FSH concentration compared to that in the vehicle-treated hamsters. These results suggest that in the hamster LHRH 1) plays a major role in maintaining FSH immunoreactivity in adenohypophysial tissue, 2) does not play a role in maintaining numbers of immunoreactive LH cells in adult adenohypophysial tissue, and 3) functions to maintain FSH synthesis at least in part in cells that contain LH.

Animals↗

PubMed1990

Luteinizing hormone (LH)-releasing hormone: chronic effects on LH and follicle-stimulating hormone cells and secretion in adult male rats.

We investigated whether chronic administration of LHRH to normal adult rats could increase the percentages of anterior pituitary gland (APG) cells that contain immunoreactive LH and/or FSH and gonadotropin secretion. Vehicle or 1 microgram LHRH was injected sc twice daily for 6 days, and rats were decapitated 16 h after the last injection. Treatment with LHRH caused nearly a doubling in the numerical density of LH and FSH cells and in the percentage of APG cells that contained LH or FSH. It also caused a shift in the gonadotroph population from LH and LH/FSH cells to LH/FSH cells. It did not change the mean size of gonadotrophs or APG weight. These changes at the light microscopic level were not accompanied by any apparent changes in LH cells at the ultrastructural level. However, they were accompanied by an approximate doubling of the basal serum LH and FSH concentrations, an increase in the APG FSH concentration, and an increase in the basal FSH release rate (measured in vitro). The results indicate that exogenous LHRH can be administered to increase numbers of gonadotrophs in the APG, synthesis of FSH in gonadotrophs, and basal serum LH and FSH concentrations.

Animals↗

PubMed1990

The response of splenic lymphocytes removed from hypophysectomized-orchidectomized hamsters to phytohemagglutinin correlates with somatic growth but not with circulating prolactin levels.

To examine the relationship between PRL and the mitogenic capacity of lymphocytes, we studied the relationships among circulating PRL levels, somatic growth, and the response of splenic lymphocytes to the mitogen phytohemagglutinin (PHA) in hamsters. In the first experiment, no differences were observed in the PHA responses of lymphocytes removed from intact or hypophysectomized-orchidectomized hamsters. No relationships were observed between circulating PRL levels and either the PHA responses or somatic growth. However, significant positive correlations were observed between the somatic growth of intact or hypophysectomized-orchidectomized hamsters and the PHA responses (r = 0.741; P less than 0.01 for intact hamsters; r = 0.642; P less than 0.01 for hypophysectomized-orchidectomized hamsters). In three subsequent experiments we tested the effects of placing muscle or hypophysial allografts in hypophysectomized-orchidectomized hamsters on somatic growth, the PHA responses, and circulating PRL levels. Neither type of allograft altered the somatic growth of hypophysectomized-orchidectomized hamsters. The hypophysial allografts did elevate serum PRL levels. In all experiments the responses of splenic lymphocytes to PHA showed a significant positive correlation with somatic growth, but not with serum PRL levels. These results minimize a role of PRL in this particular lymphocyte response. The results suggest that a strong correlation exists between mechanisms responsible for somatic growth in hypophysectomized-orchidectomized hamsters and the immune status, as determined by the response to PHA, of the animals. This relationship also may exist in intact hamsters.

Adrenal Glands↗

PubMed1989

Effects of corticotrophin-releasing hormone on corticotrophs in anterior pituitary gland allografts in hypophysectomized, orchidectomized hamsters.

We investigated the effects of corticotrophin-releasing hormone (CRH) on the percentage of anterior pituitary gland (APG) cells which are corticotrophs as well as the size and shape of corticotrophs. Pituitary glands were removed from 7-week-old male hamsters and placed beneath the renal capsules of hamsters that had been hypophysectomized and orchidectomized 3 weeks previously. Beginning 6 days after each host had received a single allograft, each was injected subcutaneously twice daily with 4 micrograms CRH or vehicle for 16 days. Six hosts in each group were decapitated 16 h after the last injection. Sections of anterior pituitary tissue were stained for ACTH and with hematoxylin. The percentage of corticotrophs among APG cells was greater in allografts exposed to exogenous CRH (approximately 20%) than in allografts exposed to vehicle (approximately 15%). Exposure to exogenous CRH increased the cross-sectional area of corticotroph cells in allografts to values greater than those measured for corticotrophs in allografts exposed to vehicle, without altering the shape of cells. Results of subsequent studies suggested that hamsters with allografts injected with vehicle do not release ACTH and that exogenous CRH causes an abrupt release of ACTH from allografts. These results indicate that CRH releases ACTH from ectopic corticotrophs and that administration of CRH can increase corticotroph size and the percentage of APG cells that are corticotrophs.

Adrenocorticotropic Hormone↗

PubMed1989

Luteinizing hormone (LH)-releasing hormone: effects of induction of LH, follicle-stimulating hormone, and prolactin cell differentiation.

We investigated the influence of LHRH on the differentiation of gonadotrophs and lactotrophs in fetal pituitary glands transplanted beneath the renal capsules of adult hypophysectomized-orchidectomized hamsters (hosts). Hypophyses were removed from hamster fetuses at a gestational age of 14 days. Some of these were immediately fixed in Bouin's solution, and others were transplanted into the hosts. The hosts were injected sc twice daily with 1 microgram LHRH or vehicle for 16 days. Six hosts in each group were killed by decapitation 16 h after the last injection. Six 14-day-old normal male hamsters (age-matched to correspond to the age of the allografts at the time of the hosts' decapitation) also were decapitated. Sections of hypophyses in situ from fetal hamsters, from 14-day-old controls, and from allografts in each group were stained for LH, FSH, or PRL and with hematoxylin. No PRL-containing cells and very few LH or FSH cells (less than 0.025% of the adenohypophysial cell population) were observed in fetal pituitary glands. In allografts from the vehicle-treated hosts, 21.1% of adenohypophysial cells contained LH, but only 1.8% contained FSH. In allografts from LHRH-treated hosts, 28.0% and 22.9% of the adenohypophysial cells contained LH and FSH, respectively. Adenohypophyses that developed for the same length of time in situ had smaller percentages of adenohypophysial cells containing LH (23.8%) and FSH (15.5%) than the LHRH-treated group. LH-containing cells in allografts in the vehicle-treated hamsters, but not in the LHRH-treated animals, were reduced in size compared to those measured in situ. The number of lactotrophs in all allografted tissue was markedly reduced compared to that of lactotrophs in situ, and injection of LHRH into hamsters with allografts did not alter the percentage of adenohypophysial cells that were lactotrophs. These results suggest that in the hamster LHRH 1) plays an important role in stimulating the formation of immunoreactive FSH in the pituitary gland, 2) can increase the number of gonadotrophs that develop during the neonatal period, and 3) plays a role in controlling the size of gonadotrophs during development. The results also suggest that the development of lactotroph cell number requires close proximity to the hypothalamus and/or exposure to a neonatal environment. We found no evidence to support the view that LHRH, LH, or FSH stimulates immunoreactive lactotroph differentiation.

Animals↗

PubMed1988

Development and retention of phenotypically specialized cells in pituitary allografts in the hamster (Mesocricetus auratus).

We used immunohistochemistry to identify cells present in pituitary allografts in the hamster. Hypophyses removed from neonatal hamsters or adenohypophyses removed from adult females were placed beneath renal capsules of hypophysectomized adult females. Serum PRL, LH, and GH concentrations were measured at two, five and eight weeks after placement of allografts. Allografts were removed after eight weeks and stained for cells containing PRL, LH, FSH, GH, or ACTH. Allografts did not release LH or GH. Those of adult adenohypophyseal tissue released significantly more PRL. The morphology of allografts of neonatal hypophyseal tissue resembled that of the adult adenohypophysis in situ. Lactotrophs, corticotrophs, somatotrophs and LH-cells were observed; very few FSH-cells were present. Allografts of adult adenohypophyseal tissue contained pituitary cells, numerous cavities, often enclosing lymphoid cells, and fibrous tissue. Atypical lactotrophs were the numerically dominant cells in these allografts; all other cells were present. The LH-cells outnumbered FSH-cells. These observations suggest that: (a) development of normal adenohypophyseal morphology can occur in an ectopic position; (b) intracellular hormones are present in cells in an ectopic site; (c) development and retention of intracellular FSH is more dependent on occupation of the normal position of the adenohypophysis than is retention of intracellular LH; and (d) release of PRL occurs from atypical cells in allografts of adult adenohypophyseal tissue.

Animals↗