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Biomedical subjects

I S King

Publications and source records attributed to I S King.

10 recordsLinked to original sources

Androgen receptor-immunoreactive cells in ram hypothalamus: distribution and co-localization patterns with gonadotropin-releasing hormone, somatostatin and tyrosine hydroxylase.

Testosterone exerts important feedback effects on the hypothalamus of the ram to influence reproductive functioning. To provide a neuroanatomical basis for understanding this androgen action, the present study has examined androgen receptor (AR) immunoreactivity within the hypothalamus and adjacent brain areas of the intact non-breeding season ram. The largest populations of AR-immunoreactive cells were detected in the medial preoptic area, infundibular and premammillary nuclei in addition to the ventromedial nucleus (VMN) where cells were found distributed throughout its medial and lateral divisions. Smaller numbers of AR-expressing cells were identified in the bed nucleus of the stria terminalis and anterior hypothalamic area (AHA) including the paraventricular, but not the supraoptic, nucleus. Double-labelling immunocytochemistry revealed the presence of AR immunoreactivity in only 2 of 460 gonadotropin-releasing hormone (GnRH) neurons. A very small population of TH-immunoreactive cells located in the lateral aspect of the AHA was found to contain ARs. Dopaminergic cells elsewhere in the hypothalamus, including the infundibular nucleus, did not display AR immunoreactivity. Nearly 50% of AR-expressing cells in the lateral VMN were immunoreactive for somatostatin while less than 5% of periventricular somatostatin neurons displayed AR immunoreactivity. These results show where ARs are expressed in the ram hypothalamus and indicate the neuroanatomical sites at which androgen may act to influence reproductive function. The absence of ARs in the neuroendocrine GnRH and tuberoinfundibular dopaminergic cells suggests that androgens do not influence the genome of these cells in any direct manner. In contrast, the somatostatin neurons of the VMN appear to be an important target for circulating androgens in the non-breeding season ram.

Animals

Increased fos expression in preoptic calcitonin gene-related peptide (CGRP) neurones following mating but not the luteinizing hormone surge in female rats.

The functional relationship between sexually dimorphic neural populations and sex differences in reproductive functioning is unclear. The present study has investigated the function of the sexually dimorphic, estrogen-receptive, calcitonin gene-related peptide (CGRP) neurones in the female preoptic area by examining patterns of Fos immunoreactivity within these cells in relation to the luteinizing hormone surge and lordosis behaviour. In the first experiment, ovariectomized rats were treated with estradiol alone or estradiol plus progesterone to induce the luteinizing hormone surge. The percentage of CGRP neurones with Fos-positive nuclei was not different in estradiol alone (18 +/- 4%) and estradiol/progesterone-treated (24 +/- 3%) rats although the number of Fos-immunoreactive cells in the medial preoptic nucleus was increased 2-fold (P < 0.01) in estrogen/progesterone-treated rats and 40 +/- 5% of luteinizing hormone-releasing hormone neurones were found to express Fos in this group. In the second experiment, ovariectomized rats were treated with estradiol and progesterone and either, mated with a single male or placed in an empty cage, for 30 min. The number of Fos-immunoreactive cells in the medial preoptic nucleus was increased 4-fold in mated rats (P < 0.01) and the percentage of CGRP neurones with Fos-positive nuclei increased from 24 +/- 3% to 38 +/- 2% (P < 0.01) in mated animals. No differences were detected in the number of luteinizing hormone-releasing hormone neurones with Fos-positive nuclei in mated and non-mated animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Biotinylated anti-progesterone monoclonal antibodies specifically target the uterine epithelium and block implantation in the mouse.

Post-coital administration of a mouse monoclonal antibody (mAb) against progesterone (DB3; IgG1) prevents pregnancy in several species. Our previous studies in mice have shown that passively transferred DB3 specifically targets the uterus before the expected time of implantation, probably through progesterone-binding sites. DB3 and two other anti-progesterone mAbs, i.e. 11/34 (IgG1) and 11/64 (IgM), were biotinylated and 9 nmol of each (a dose known to reduce pregnancy rate by greater than 80% with unconjugated mAbs) was injected into BALB/c female mice 32 h post coitum (p.c.). The biotin-mAb complexes were highly effective in blocking pregnancy (83-100%) compared with control animals that had received the biotinylated MOPC21 myeloma protein P3 (IgG1). Using the streptavidin-FITC (fluorescein isothiocyanate) reporter complex or second-stage anti-biotin-FITC antibodies, biotinylated conjugates of DB3, 11/34 and 11/64 were specifically localized on the uterine luminal epithelium at 68 h p.c. (i.e. 36 h after i.p. injection). Neither P3-biotin-treated pregnant mice nor biotinylated anti-progesterone mAb-treated pseudopregnant females showed a positive reaction. In addition, the localization of biotin-conjugated anti-progesterone mAbs could be blocked by absorption of the antibodies with free progesterone or progesterone conjugates prior to injection. These results show that localization to the uterine epithelium occurs with different anti-progesterone mAbs, and that this phenomenon is probably associated with progesterone-binding sites on the luminal epithelium.

Animals

Effect of diet upon enterocyte differentiation in the rat jejunum.

Positional and temporal correlates for the development of microvillus membranes and for two of the hydrolytic enzymes they contain have been determined and compared with the ability of enterocytes to transport valine during migration from crypt base to villus tip in jejunal tissue taken from rats maintained on diets containing different amounts of protein. Microvillus elongation and the appearance of both aminopeptidase N (APN) and isomaltase (IM) activities reached maximal rates of expression in enterocytes located 16 +/- 5 micron from the crypt-villus junction. This close positional correlation was not found for the later development of the valine transport function. Feeding rats isoenergetic diets containing 20% instead of 5% protein caused significant increases in both villus height and crypt depth without changing the positional correlations described above. The maximal rates for microvillus elongation and APN and IM appearance were greater and occurred earlier in enterocytes taken from rats fed a high-protein diet. The time of onset and capacity to transport valine were found to be closely correlated for rats maintained on high- and low-protein diets. The ratio of APN to IM activity in fully differentiated enterocytes was either 0.7 or 1.2 depending on whether rats had been fed a low- or high-protein diet. The maximal length of microvillus membranes in fully differentiated enterocytes from rats on a low-protein diet was 1.4 times that found in rats maintained on a high-protein diet. Possible ways in which the microvillus membrane structure of enterocytes, enzyme activity and the ability to transport amino acids might be controlled are discussed. Relative estimates are also made of the probable effects that changes in diet will have on the capacity of the intestine to digest and absorb nutrients.

Aminopeptidases

Kwashiorkor: an electron histochemical study of the hair shaft.

Previous studies on hair in kwashiorkor have been equivocal regarding the normal or abnormal incorporation of amino acids into the hair shaft. In this study we have applied a sensitive electron histochemical technique to transverse sections of scalp hair from children with severe kwashiorkor in order to study its ultrastructure and the incorporation of sulphur-containing amino acids (cystine). We have found no evidence to support the idea that a protein deficient diet in the weaning or post-weaning period directly affects the distribution and relative amounts of sulphur-containing amino acids in the hair shaft in this disease.

Child

Cellular distribution of neutral and basic amino acid transport systems in rabbit ileal mucosa.

1. An autoradiographic technique is described whereby the cellular location of tritiated amino acids can be determined following uptake by rabbit ileal mucosa. 2. Stirring solutions in contact with the intestinal mucosa during measurement of rapid influx changes the quantity, but not the distribution, of alanine taken up by the tissue. 3. Conditions predicted to favour either a high affinity system (Ly1) or a low affinity system (Ly2) were used to measure lysine distribution following uptake. Maximal uptake for both transport systems occurred in fully differentiated enterocytes at the tips of villi. Initial maturation of the Ly1 system, which was slow, was followed by a rapid phase of development. The Ly2 system lacked this rapid phase of late development. 4. The cellular distribution of alanine entering on a low affinity Na-independent neutral amino acid carrier closely resembles that determine for Ly1 system for lysine entry. 5. Arginine is a potent inhibitor of lysine uptake through the Ly2 system. Little or no diffusion of lysine appears to take place into rabbit ileal enterocytes. 6. The different distribution of the high and low affinity systems for lysine transport provides further support for their independent existence. It also suggests that more than one message exists fo the switching on of amino acid transport function in differentiating enterocytes.

Alanine

Cell proliferation in follicle-associated epithelium of mouse Peyer's patch.

Follicle-associated epithelium (FAE) from the mouse Peyer's patch consists of a dome part containing goblet as well as columnar epithelial cells and a crypt part, containing Paneth as well as goblet and undifferentiated epithelial cells. The proportions of columnar epithelial, goblet and Paneth cells in the FAE are similar to those found in normal crypt/villus complexes. Mitosis is confined to the crypts of the FAE. The cell-renewal time for FAE columnar epithelial cells, estimated from experiments using tritiated thymidine, is approximately 60 hours. Large numbers of lymphoid cells are also present in the FAE, particularly in regions distal to the crypts. The similarity between follicle-associated and villus epithelial structure is discussed, together with its possible relevance to "M" cell formation within the FAE.

Animals