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Biomedical subjects

B Colenbrander

Publications and source records attributed to B Colenbrander.

At least 109 records · Page 6Linked to original sources
PubMed1990

Isolation and characterization of boar spermatozoa with and without a cytoplasmic droplet.

1. Boar semen was separated on a Percoll density gradient into three populations; a low-density band of immature sperm cells containing a cytoplasmic droplet and a high-density doublet band formed by spermatozoa without a cytoplasmic droplet. 2. In these three cell populations four acid hydrolases were determined, viz. (1) alpha-glucosidase; (2) alpha-mannosidase; (3) beta-galactosidase; (4) beta-hexosaminidase. 3. The release of the hydrolases (1), (2) and (3) from cytoplasmic droplet containing spermatozoa was stimulated whereas the release of beta-hexosaminidase was inhibited by calcium ions. 4. The results suggest that acid alpha-glucosidase, alpha-mannosidase and beta-galactosidase are situated in the acrosome whereas acid beta-hexosaminidase is localized predominantly in the cytoplasmic droplet of boar spermatozoa. 5. We conclude that beta-hexosaminidase should prove useful as a biochemical marker for cytoplasmic droplet containing spermatozoa and hence for the number of immature sperm cells in boar semen.

Acrosome↗
PubMed1989

Metabolic dissimilarity between (9,12,12-2H) cortisol and natural cortisol in vivo. Can deuterated cortisol be used for the measurement of the urinary cortisol production rate?

The metabolism of deuterated cortisol (9,12,12,-2H)cortisol, 2H3-F) was compared to that of radioactive cortisol (3H2-F) and natural cortisol, when these three compounds were administered simultaneously to an adrenalectomized piglet. The relative isotope dilution of tritium was determined from the specific activities of the main urinary neutral cortisol metabolites, tetrahydrocortisone (THE) and tetrahydrocortisol (THF), normalized to that of the cortisol mixture administered. To obtain a comparison of the isotope dilution of deuterium in the metabolites THE and THF to that in the cortisol mixture, the three steroids were converted to the common oxidation product 11-oxo-aetiocholanolone, and derivatized to the methoxime-tert-butyl-dimethylsilyl ether. The relative 2H-isotope dilution then was measured by gas chromatography/mass spectrometry. It was found that the specific activity of THE in the cumulative urine collections was similar to that of the cortisol mixture administered; the two-day value was, however, less. The specific activity of THF was slightly but significantly smaller than 1 (approximately 0.9) at all times. The relative 2H-isotope dilution in THE was slightly but significantly larger than one (approximately 1.1) at all times, whereas that in the THF was larger than 1.0 at 9 and 32 h or equal to 1.0 at 20 and 47 h of urine collection. When comparing the metabolism of the two tracer cortisol species the quotient of the 3H- and the 2H-isotope dilutions in THE and THF was smaller than 1.0. It can be concluded that (2H3)cortisol may be used for the determination of the cortisol production rate.

Adrenalectomy↗
PubMed1989

Effects of pure FSH and LH preparations on the number and function of Leydig cells in immature hypophysectomized rats.

The effects of pure FSH and/or LH preparations on the number of Leydig cells and their function in immature hypophysectomized rats have been investigated. As a result of hypophysectomy at the age of 17-18 days, the number of recognizable Leydig cells per testis decreased, as did the steroidogenic capacity in vivo and in vitro. Treatment with 64 micrograms FSH on both 22 and 23 days of age, did not affect the number of recognizable Leydig cells. In contrast, two injections of LH (10 micrograms) caused a sixfold increase in the number of Leydig cells, but had a negative effect on spermatogenesis. These stimulatory and inhibitory effects of LH diminished when FSH was added. Treatment with FSH for 7 days caused a twofold increase in the number of Leydig cells when compared with hypophysectomized controls. 3 beta-Hydroxysteroid dehydrogenase (3 beta-HSD) and esterase activity in Leydig cells also increased under the influence of FSH. The pregnenolone production per Leydig cell in the presence of 5-cholesten-3 beta,22(R)-diol (22R-hydroxycholesterol) as substrate showed a sevenfold increase. Plasma testosterone levels 2 h after injection of human chorionic gonadotrophin in intact rats and hypophysectomized FSH-treated rats were the same. Following LH treatment for 7 days, the number of Leydig cells proved to be 11 times higher, and 3 beta-HSD and esterase activity were not different from intact controls. The testicular pregnenolone production was four- to fivefold higher when compared with untreated hypophysectomized rats. However, pregnenolone production per Leydig cell in LH-treated rats was only slightly different from the hypophysectomized controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Immunohistochemical and biochemical analysis of the development of the noradrenaline- and adrenaline-storing cells in the adrenal medulla of the rat and pig.

The development of the noradrenaline (NA)- and adrenaline (A)-storing cells was examined in the adrenal gland of pre- and postnatal rats and pigs. Cryostat sections were immunostained with antibodies to NA and A. Amine levels were estimated in homogenates of adrenals by ion-pair reversed-phase liquid chromatography with electrochemical detection. 1. In adult animals separate NA- and A-storing cells were found. In the rat A-cells comprised about 80% of the parenchyma of the adrenal medulla. NA-cells, the remaining 20%, were randomly arranged in clusters. In the pig, in contrast, the A- and NA-storing cells were equally distributed, with spherical clusters of NA-cells were surrounded by A-cells. 2. In the earliest developmental stages examined (16th day of gestation in the rat, and 42nd day in the pig) the adrenals only contained NA-immunoreactive cells. In the rat separate NA- and A-storing cells were first noticed 2 or 3 days after birth, whereas in the pig separated cells were already present at the 56th prenatal day (full term at 114th day). 3. In both rat and pig adrenals dopamine (DA), NA and A increased in amount rapidly during development. In the rat this change mainly took place after birth (an increase of 13 times from the 17th day of gestation, and a 100-fold increase from birth to adult age). In porcine adrenal increase of the total amount of amines mainly occurred before birth (200 times between the 42nd day of gestation and birth, whereas only 10 times after birth). 4. In both species DA-levels remained low during pre- and postnatal development. On the contrary, the relative concentrations of NA decreased while that of A increased correspondingly. Again, species differences were noticed: in the rat NA decreased from 90% (17th day of gestation) via 35% (just before birth) to 20% at adult age, while the porcine adrenal showed a more gradual decrease, i.e., from 90% (42nd day of gestation) to 70% (birth) ending up with 50% in the adult stage. 5. The immunohistochemical and biochemical data indicate that, in rat adrenal medulla three phases of development can be distinguished. First, up to the 18th day of gestation, medullary cells synthesize and store only NA. Second, from the 18th day to 2 or 3 days after birth, NA and A are synthesized and stored in a single cell type ("mixed cell type"), and third, NA and A are localized in separate cell types.(ABSTRACT TRUNCATED AT 400 WORDS)

Adrenal Cortex Hormones↗
PubMed1988

Familial male pseudohermaphroditism and testicular descent in the racoon dog (Nyctereutes).

Sexual differentiation was investigated in familial male pseudohermaphroditism in Nyctereutes procyonoides (Canidae). In intersex males, development of external genital organs and prostate glandular tissue was severely disturbed; Wolffian (mesonephric) duct derivatives developed prepubertally but were absent in some adults. Müllerian (paramesonephric) duct regression was complete. Testicular descent was undisturbed. Male/female sex differences in plasma testosterone, 5 alpha-dihydrotestosterone, and luteinizing hormone concentrations were present. Intersex plasma hormone concentrations were within the normal male range. The concentration of androgen receptors in pubic skin was similar in male, female, and intersex animals and no significant differences in affinity for the ligand were detected. It was concluded that in intersex animals androgen-dependent virilisation was deficient despite the presence of androgens and androgen receptors and that this condition had not affected gubernaculum development and testicular descent.

Animals↗
PubMed1988

Are (13C)cortisol and (3H)cortisol metabolized identically to natural cortisol in adrenalectomized piglets?

Adrenalectomized piglets were intravenously administered a mixture of (13C4)cortisol and (3H)cortisol and natural cortisol to determine if the two tracers are metabolized identically to natural cortisol. Urine was collected after 0.5, 1.0, 1.5 and 2.0 days and the isotope dilution was measured in the four major urinary cortisol metabolites, namely tetrahydrocortisone (THE), tetrahydrocortisol (THF), alpha- and beta-cortolone in the cumulative urines. In contrast to other studies, because of the sensitivity of the method used to measure the 13C4 enrichment, non-cumulative urine collections were also analysed. Quantification of the 13C4 isotope enrichment was carried out by gas chromatography/mass spectrometry with selected ion monitoring. The specific activities of the metabolites from the cumulative urine collections were determined by high-performance liquid chromatography and scintillation counting. Small secondary isotope effects seemed to occur during the metabolism of (13C4)cortisol, as a decrease in isotope enrichment in all four metabolites was measured. These effects were easily observed with alpha- and beta-cortolone isolated from the cumulative urine collections; the enrichment decreased by 19% and 14%, respectively. The lowering in isotope dilution in THE observed in the 2.0 day cumulative urine collection in piglets 1 and 2 were 4% and 3%, respectively. A lowering in isotope dilution in THF in the 2.0 day cumulative urine collection could be observed in piglet 2, namely 7%, but no change in isotope dilution could be seen in piglet 1. These secondary isotope effects could only be observed in the 2 days cumulative urine, and not in the cumulative urines collected over shorter times. The non-cumulative urines collected at half-day periods showed a significant decrease in isotope dilution in THE and THF isolated from the urine collected after 1 day. No statistically significant isotope effects were observed with the metabolism of (3H)cortisol, except at 0.5 day when the specific activity in the cortolones was lower and that in THF was higher. However, at 0.5 day with THE and 1.0 day with THF and the cortolones the specific activities remained approximately 6% higher than that administered in the cortisol. Secondary isotope effects with tritiated cortisol may have occurred but because of the relatively large imprecision of the measurement (SD = 3-4% with THE and THF and the cortolones (SD approximately 8%) compared to the measurements of the 13C4 enrichment (SD approximately 2%) these effects could not statistically be proven.(ABSTRACT TRUNCATED AT 400 WORDS)

Adrenalectomy↗
PubMed1988

Prolactin in the developing pig.

Prolactin (PRL) was determined in plasma of fetal pigs from 40 days post coitum (d.p.c.) onwards. Values increased in the last 3 wk of the gestational period and reached 10 ng/ml at term. Immunoreactive lactotropes could be identified as early as 31 d.p.c. Lactotrope development appeared to be biphasic, with no apparent increase in cell number in the 51-75 d.p.c. period. Sex differences in volume percentage of lactotropes and plasma PRL levels were observed in the 71-75 d.p.c. period. Pituitary PRL content increased 20-fold between 95 d.p.c. and term and 10-fold between birth and 6 wk. Volume percentage of lactotropes did not change from birth to 6 wk. In the postnatal period, plasma PRL concentrations were high in the first 1-2 wk and they decreased thereafter. Lower stable values were observed until about 10 wk, when PRL concentrations began to increase. Prepubertal peak levels were observed in the 10- to 16-wk period and a pubertal peak was observed at 20-22 wk. No significant differences between plasma PRL profiles of males and females were observed in the postnatal period. Castrated males, however, had consistently lower plasma PRL concentrations than intact boars. Cryptorchid boars had PRL concentrations comparable to intact boars. The PRL profile, in relation to the pattern of gonadal development, suggests a role for PRL in gonadal development of the pig.

Animals↗
PubMed1987

Feedback regulation of gonadotropic hormone secretion in neonatal pigs.

The effect of castration and of administration of charcoal-treated porcine follicular fluid (pFF) containing inhibin-like activity on plasma concentration of gonadotropic hormones was studied in neonatal pigs. Plasma follicle-stimulating hormone (FSH) concentration averaged 25.1 +/- 1.5 ng/ml (mean +/- SEM) in 1-wk-old females and gradually declined to 20.2 +/- 0.7 ng/ml 6 wk later. Ovariectomy did not significantly influence plasma FSH concentration. In males, concentration averaged 8.0 +/- 0.7 ng/ml before castration but rose significantly within 2 days after castration. Injection of luteinizing hormone-releasing hormone (LHRH) did not influence plasma FSH concentrations in intact males, but did in females and in 7-wk-old males castrated at 1 wk. Plasma luteinizing hormone (LH) concentrations in 1-wk-old females (2.2 +/- 0.4 ng/ml) gradually declined and were not influenced by castration. Concentrations of plasma LH in 1-wk-old male piglets (2.8 +/- 0.7 ng/ml) were not significantly influenced by castration within 2 days but were significantly higher 6 wk later. LHRH induced a significant rise in plasma LH concentrations in all animals. Injection of pFF resulted in a decline of plasma FSH concentrations in intact and castrated males and in intact females, but did not influence plasma LH concentrations. These data demonstrate a sex-specific difference in the control of plasma FSH, but not in plasma LH concentration in the neonatal pig. Plasma FSH concentrations, but not plasma LH concentrations, are suppressed by testicular hormones in 1-wk-old piglets. Plasma FSH concentrations can be suppressed in both neonatal male and female pigs by injections of pFF.

Aging↗
PubMed1987

The ultrastructure of normal fetal and neonatal pig testis germ cells and the influence of fetal decapitation on the germ cell development.

The development of germ cells in the male pig was investigated ultrastructurally in normal and decapitated fetuses. The age ranged respectively from 30 days p.c. till one month after birth and from 52 days p.c. until birth. The ultrastructural organization of the germ cells changes dramatically between 30 days p.c. and 52 days p.c. which coincides with the formation of 'true' sex cords. From 52 days p.c. onwards the morphology is rather stable: cells show a 'hydrated' appearance and typical cell bridges. There is no obvious difference in the ultrastructure of germ cells in decapitated animals, their normal littermates and control animals. Therefore germ cell development in the pig is likely to be insensitive to gonadotropins during the fetal period. The development of pig germ cells follows closely the pattern described for several species. Quantitatively there is an increase in the ratio of germ cell/Sertoli cell per cross sectional diameter in the decapitated animals.

Animals↗
PubMed1986

Changing basal and stimulated activity of the renin-angiotensin system during the second half of gestation in the anaesthetized piglet.

Concentrations of components of the renin-angiotensin system were studied in eighty anaesthetized pig fetuses aged between 57 and 112 d of gestation (term = 114 +/- 1 d). Plasma renin concentration rose from a mean value of 3.2 +/- 0.4 ng X ml-1 X h-1 (mean +/- S.E.M.) at 57 d to 17.5 +/- 2.7 ng X ml-1 X h-1 at 93 d of gestation and thereafter declined to 5.0 +/- 1.6 ng X ml-1 X h-1 just prior to term. There was no distinct pattern of change in plasma renin substrate concentrations during gestation; the concentration near term (0.33 +/- 0.10 micrograms X ml-1) was not different from that at 57 d (0.50 +/- 0.18 micrograms X ml-1) or that at 93 d (0.31 +/- 0.08 micrograms X ml-1) of gestation. Plasma angiotensin II concentrations increased from 33 (log10 (mean +/- S.E.M.) = 1.554 +/- 0.050) pg X ml-1 at 60 d to 232 (2.365 +/- 0.225) pg X ml-1 at 105 d of gestation, after which it decreased to 50 (1.699 +/- 0.100) pg X ml-1 in 20-d-old neonatal piglets. Haemorrhage of 10-20% estimated blood volume produced no consistent changes in plasma renin substrate concentrations, and inconsistent changes in renin and angiotensin II concentrations at 85-90 d of gestation, but a consistent increase in both renin and angiotensin II concentrations thereafter. These results indicate that the components of the renin-angiotensin system are present in the circulation of the pig fetus, and that their responses to stimuli develop during the second half of gestation.

Anesthesia↗
PubMed1986

Ontogeny of hypothalamic pituitary function in the pig: pituitary thyrotrophin in the fetus and neonate.

Thyrotrophin in individual pituitaries obtained from fetal and prepubertal pigs was quantified by homologous radioimmunoassay (RIA) and heterologous radioreceptor assay (RRA). Relative evolution of pituitary TSH contents and concentrations with age were in good agreement as measured by both assay systems although the quantity of TSH detected by RRA appeared consistently lower than that measured by RIA. Thyrotrophin was first detected in pituitaries of fetal pigs at day 75 of gestation. Thereafter the pituitary content of TSH increased to approximately 45 micrograms/pituitary in the oldest group tested (6 weeks of age). The pituitary TSH concentration rose sharply until birth (114 +/- 1 day post coitum) and thereafter remained increased at a concentration of approximately 400 ng/mg wet weight.

Animals↗
PubMed1986

Kinetic measurement of the urinary production rate of cortisol in male piglets: is the prerequisite 'collection until all label has disappeared' necessary?

Urinary cortisol production rate (CPR) was calculated by two different methods in five male piglets (about 3 kg bodyweight) injected i.v. with 40-120 kBq tritiated cortisol ([3H]F. After administration of [3H]F, urine was obtained from four consecutive collections for the following 2 days, during which 80-100% of the label was recovered. Total radioactivity in the urine was measured and used to calculate the total rate constant of 0.115 +/- 0.011 h-1 and, from this, the mean biological half-life (t1/2) of 6.0 +/- 0.6 h (S.D.; n = 4). It was found that the mass ratio of the two principal urinary cortisol metabolites tetrahydrocortisone (THE) and tetrahydrocortisol (THF) was strikingly less than 1.0 (0.4 +/- 0.1; n = 14), which is the reverse of that observed in older pigs, neonatal infants and man. To calculate CPR conventionally, the cumulative specific activities of THE and THF were calculated for the 2-day period of urine collection. The apparent mean CPR values on the basis of THE and THF were calculated as 11.5 +/- 1.6 (n = 5) and 12.8 +/- 3.3 (n = 5) mumol/day respectively, and 12.1 +/- 1.4 (n = 5) mumol/day for the average of THE and THF. The second method for calculating CPR consisted of determining the masses of THE and THF (mumol) per fraction of dose (m/fd) (fd refers to the ratio of radioactivity in the metabolite and dose) at different times after administration of [3H]F. The calculated m/fd values, which are synonymous with the dose divided by the specific activities of the metabolites, and the different times of urine collection were analysed by linear regression. The resulting slope is equal to the CPR. The CPR derived by this method for the average of THE and THF, 10.1 +/- 0.91 mumol/day was significantly (P less than 0.014) lower than that derived conventionally, 12.1 +/- 1.40 mumol/day. This second method may be used when CPR is determined in neonatal infants by means of nonradioactive, deuterated or 13C-enriched cortisol, where the extent of negative feedback by the relatively high dose of exogenous steroid on cortisol secretion must be kept as low as possible. This method also allows urine collections to be used at times when the tracer is still being excreted.

Animals↗
PubMed1986

Some histochemical and ultrastructural observations on the early foetal pig testis.

Testes of foetal pigs between 26 to 35 days post coitum (p.c.) were investigated histochemically and ultrastructurally. Diaphorase and delta 5-3 beta-hydroxysteroid dehydrogenase activities were studied using, respectively, NADH and pregnenolone and dihydroxy androsterone as substrates. Ultrastructurally, attention was focused on the development of mesenchymal cells and on the sustentacular cells in the primitive sex cords in an attempt to detect the origin of Leydig cells. Histochemically there is a concentration of activity toward the interstitium with increasing age. Also the reactions increase in intensity. Ultrastructurally no evidence for Leydig cell development from Sertoli cells could be observed. Mesenchymal cells between the sex cords show a development toward Leydig cells. This is absent in mesenchymal cells in the future tunica albuginea. Before 30 days p.c. no 'true' Leydig cells can be observed morphologically. The role of the rough endoplasmic reticulum/mitochondrial complex, which is present in many mesenchymal and sustentacular cells, is discussed.

3-Hydroxysteroid Dehydrogenases↗
PubMed1985

The ontogeny of hypothalamic pituitary function in the pig. I. Pituitary LH and FSH in the fetus and neonate.

Pituitaries were collected from fetal and postnatal pigs from day 55 p.c. until 6 weeks after birth at closely spaced intervals. LH and FSH in individual pituitaries were quantified by both homologous RIA and homologous RRA. No significant difference was found between results obtained by RIA and RRA. Both LH and FSH are first detected by RIA and RRA in the porcine fetal pituitary at 75 days p.c. Thereafter both LH and FSH pituitary content rises until term. LH pituitary concentration in both male and female fetuses and FSH pituitary concentration in males exhibit a peak just before birth. FSH pituitary concentration in females rises until birth and thereafter remains elevated. A statistically significant sex difference was found postnatally with regard to FSH content and concentration but not for LH.

Animals↗
PubMed1985

Development of pituitary gonadotropic cells in the pig fetus and the effect of luteinizing hormone-releasing hormone administration.

Pituitary gonadotropic development in fetal pigs has been studied immunocytochemically. From 50 days postcoitum (p.c.) until 90 days p.c. the volume density of luteinizing hormone (LH) immunoreactive cells increased from 0.12% to 0.61% of the glandular mass of the anterior part of the pituitary gland. From 90 to 100 days p.c. a steep increase to 4.5% was found. A single injection of long-acting luteinizing hormone-releasing hormone (LHRH) to the fetus at 75 days p.c. inhibited this developmental pattern. Pulsatile administration of LHRH agonist to fetuses from 70 to 73 days p.c. had no significant effect on the volume density of LH immunoreactive cells, whereas pulsatile administration from 70 to 80 days p.c. and from 95 to 105 days p.c. inhibited the normal increase of this parameter. It is concluded that from 70 days p.c. onwards the development of fetal gonadotropic cells can be manipulated by exogenous LHRH.

Animals↗
PubMed1985

Effect of somatostatin and thyrotrophin-releasing hormone on the levels of growth hormone in the circulation of the chronically catheterized pig fetus in utero.

The effects of i.v. administration of thyrotrophin-releasing hormone (TRH) and of somatostatin on circulating plasma levels of porcine GH in the chronically catheterized pig fetus have been examined. Growth hormone levels increased markedly (P less than 0.01) following TRH administration, but there was no change in thyroxine levels by 1 h after treatment. Administration of somatostatin caused a significant (P less than 0.05) decrease in mean GH levels, but the response was variable between pigs. Saline administration had no significant effect on GH levels. These results suggest that the mechanisms regulating postnatal GH release are present in the fetal pig, but may not be fully developed 8-12 days before delivery.

Animals↗