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Results for “Functional differentiation”

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At least 541 records · Page 30Linked to original sources
PubMed2006

Differential functions of the Aurora-B and Aurora-C kinases in mammalian spermatogenesis.

The Aurora kinases are cell cycle-regulatory serine-threonine kinases that have been implicated in the function of the centrosomes, kinetechores, chromosome dynamics, and cytokinesis. In comparison with other tissues, there are high levels of expression of Aurora-B and -C in testis. What their respective roles in mammalian spermatogenesis are is an open question. Here we describe the expression and distribution patterns of the three kinases in mouse testis using in situ hybridization and immunohistochemistry. Importantly, the localization of Aurora-B is tightly regulated during spermatogenesis, whereas Aurora-C expression appears to be testis specific. To address the function of Aurora-B in spermatogenesis, we have generated transgenic mice using a pachytene-stage-specific promoter driving the expression of either wild-type Aurora-B or an inactive form of the kinase. Expression of the inactive Aurora-B results in abnormal spermatocytes, increased apoptosis, spermatogenic arrest, and subfertility defects. The function of Aurora-C may also be targeted in the Aurora-B transgenic mutants. To address the function of Aurora-C in testis, we generated Aurora-C knockout mice by homologous recombination. Remarkably, Aurora-C null mice were viable, yet the males had compromised fertility. Aurora-C mutant sperm display abnormalities that included heterogenous chromatin condensation, loose acrosomes, and blunted heads. These findings indicate that Aurora-B and Aurora-C serve specialized functions in mammalian spermatogenesis.

Animals↗
PubMed2003

Differential function of RNCAM isoforms in precise target selection of olfactory sensory neurons.

Olfactory sensory neurons (OSNs) are individually specified to express one odorant receptor (OR) gene among approximately 1000 different and project with precision to topographically defined convergence sites, the glomeruli, in the olfactory bulb. Although ORs partially determine the location of convergence sites, the mechanism ensuring that axons with different OR identities do not co-converge is unknown. RNCAM (OCAM, NCAM2) is assumed to regulate a broad zonal segregation of projections by virtue of being a homophilic cell adhesion molecule that is selectively expressed on axons terminating in a defined olfactory bulb region. We have identified NADPH diaphorase activity as being an independent marker for RNCAM-negative axons. Analyses of transgenic mice that ectopically express RNCAM in NADPH diaphorase-positive OSNs show that the postulated function of RNCAM in mediating zone-specific segregation of axons is unlikely. Instead, analyses of one OR-specific OSN subpopulation (P2) reveal that elevated RNCAM levels result in an increased number of P2 axons that incorrectly co-converge with axons of other OR identities. Both Gpi-anchored and transmembrane-bound RNCAM isoforms are localized on axons in the nerve layer, while the transmembrane-bound RNCAM is the predominant isoform on axon terminals within glomeruli. Overexpressing transmembrane-bound RNCAM results in co-convergence events close to the correct target glomeruli. By contrast, overexpression of Gpi-anchored RNCAM results in axons that can bypass the correct target before co-converging on glomeruli located at a distance. The phenotype specific for Gpi-anchored RNCAM is suppressed in mice overexpressing both isoforms, which suggests that two distinct RNCAM isoform-dependent activities influence segregation of OR-defined axon subclasses.

Animals↗
PubMed1982

Dissociation of cytological and functional differential in virgin mouse mammary gland during inhibition of DNA synthesis.

Epithelial cells in mammary gland explants from mice assume a secretory appearance and synthesize the milk proteins, casein and alpha-lactalbumin, when cultured in the presence of insulin, hydrocortisone and prolactin. In cells from the glands of mature virgin animals such syntheses are known to require DNA synthesis. Addition of cytosine-beta-D-arabinofuranoside to the explant cultures suppresses both hormonally induced DNA synthesis and enhanced production of milk protein. To determine the level at which this block in terminal differentiation occurs, epithelial cell pellets were prepared from virgin mouse mammary gland explants cultured with various combination of insulin, hydrocortisone and prolactin, and subsequently examined by light and electron microscopy. We observed that the epithelial cells cultured in the presence of all three hormones developed fully, cytologically and ultrastructurally, even in the absence of DNA synthesis in vitro. Likewise, these cells were able to incorporate [3H]uridine into RNA efficiently and to incorporate amino acids into acid-precipitable polypeptides at levels equivalent to the untreated controls. However, immunoprecipitation of newly synthesized casein peptides showed that no new synthesis of casein occurred in cells prevented from synthesizing DNA. These data show uncoupling of cytological development and synthesis of milk protein in mammary explants from mature virgin mice inhibited from synthesizing DNA.

Animals↗
PubMed1982

Functional differentiation of enterocytes in the follicle-associated epithelium of rat Peyer's patch.

The ability of the follicle-associated epithelium (FAE) of rat Peyer's patch to accumulate valine has been measured using a new technique of autoradiographic analysis. Maximum uptake of valine was achieved by enterocytes present in the dome region of the FAE. Valine uptake was not seen in enterocytes present on the lower slopes of the FAE or in follicle-associated crypts. The ability of the FAE to absorb valine was generally much less than that seen in enterocytes present on adjacent villi. The main reason for this discrepancy lay in the apparent inability of the FAE to develop a second phase of amino acid transport similar to that seen in villus enterocytes. It is suggested that this failure results from some unexplained interaction taking place between the FAE and its underlying lymphoid tissue.

Animals↗
PubMed1985

[The biocellular effect of thyroid hormone on functional differentiation of porcine granulosa cells in culture].

To elucidate if the thyroid hormone acts directly on the ovary, the biocellular effect of L-thyroxine (T4) on porcine granulosa cells cultured in vitro was investigated. Monolayer cultures of porcine granulosa cells obtained from small (1 approximately 2 mm), medium (3 approximately 5 mm) or large (6 approximately 11 mm) follicles were carried out in the presence of porcine FSH (100 ng/ml). Concomitant treatment with T4 promoted FSH-dependent morphological luteinization, i.e. alteration of immature granulosa cells obtained from small follicles to epithelioid form. T4 also increased FSH-stimulated induction of hCG/LH receptor on immature granulosa cells. Furthermore, T4 augmented FSH-mediated production of progesterone and estradiol by immature granulosa cells cultured in vitro. The concentration of T4 to produce the maximal stimulatory effect was 10-7 M, demonstrating that optimal concentration of thyroid hormone is required for the expression of this stimulatory action. Since T4 alone demonstrated no effect on the differentiation of porcine granulosa cells and all the stimulatory effect of T4 seems to have a permissive action on FSH-induced granulosa cell luteinization. Although insulin showed a similar effect on porcine granulosa cells, no stimulation of estradiol production by porcine granulosa cells was observed with insulin in the culture system used in this study. These results suggest that the thyroid hormone acts directly on the ovary and plays an important role in modifying the FSH-dependent cellular differentiation of immature granulosa cells.

Animals↗
PubMed1989

Identification and partial characterization of mesenchyme-derived growth factor that stimulates proliferation and inhibits functional differentiation of mouse mammary epithelium in culture.

The effect of mesenchyme on both proliferation and differentiation of mammary epithelial cells was investigated in a primary cell culture system. Mammary cells cultured on collagen gel for 4 days produced casein in response to the synergistic action of insulin, cortisol, and prolactin. When mammary epithelial cells were co-cultured with fibroblasts derived from three different kinds of fetal mesenchymal tissues, casein production was suppressed. The addition of conditioned media obtained from cultures of these mesenchymal cells stimulated DNA synthesis and reduced casein synthesis in a dose-dependent fashion in the cultured mammary cells. Although such biological actions are similar to those of epidermal growth factor (EGF), the capability to compete with EGF for EGF receptor was not found in this conditioned medium. Sephadex G-200 column chromatography revealed that molecular weight of the peak which has these biological activities was around 100,000. These results indicate that fetal mesenchymal cells secrete a substance(s) which has a stimulatory effect on proliferation and an inhibitory effect on differentiation of mammary epithelial cells.

Animals↗
PubMed2006

A stable analogue of glucose-dependent insulinotropic polypeptide, GIP(LysPAL16), enhances functional differentiation of mouse embryonic stem cells into cells expressing islet-specific genes and hormones.

Embryonic stem (ES) cells can be differentiated into insulin-producing cells by conditioning the culture media. However, the number of insulin-expressing cells and amount of insulin released is very low. Glucose-dependent insulinotropic polypeptide (GIP) enhances the growth and differentiation of pancreatic beta-cells. This study examined the potential of the stable analogue GIP(LysPAL16) to enhance the differentiation of mouse ES cells into insulin-producing cells using a five-stage culturing strategy. Semi-quantitative PCR indicated mRNA expression of islet development markers (nestin, Pdx1, Nkx6.1, Oct4), mature pancreatic beta-cell markers (insulin, glucagon, Glut2, Sur1, Kir6.1) and the GIP receptor gene GIP-R in undifferentiated (stage 1) cells, with increasing levels in differentiated stages 4 and 5. IAPP and somatostatin genes were only expressed in differentiated stages. Immunohistochemical studies confirmed the presence of insulin, glucagon, somatostatin and IAPP in differentiated ES cells. After supplementation with GIP(LysPAL16), ES cells at stage 4 released insulin in response to secretagogues and glucose in a concentration-dependent manner, with 35-100% increases in insulin release. Cellular C-peptide content also increased by 45% at stages 4 and 5. We conclude that the stable GIP analogue enhanced differentiation of mouse ES cells towards a phenotype expressing specific beta-cell genes and releasing insulin.

Animals↗
PubMed1989

Amino-acid sequences and functional differentiation of hemoglobins A and D from swift (Apus apus, Apodiformes).

The blood of the adult swift contains one major (HbA = alpha 2A beta 2) and two minor components (HbD = alpha 2D beta 2 and HbD'). The components were separated by FPLC with a TSK SP-5 PW-column in phosphate buffers, and were eluted with a linear NaCl gradient. HbD' could be detected only in freshly prepared hemolysates with the sensitive FPLC separation method. The globin chains were separated on a cation exchanger (CM-cellulose), the tryptic peptides by HPLC with a RP-2 LiChrosorb column. Their amino-acid sequences were determined by automatic Edman degradation with the film- or gas-phase method. For the alpha A-, alpha D- and beta-chains, peptide alignment was achieved by homologous comparison with the corresponding chains of the greylag goose (Anser anser). The structural significance of the substitutions was examined with the aid of molecular graphics. The oxygen-binding properties of the stripped hemolysate and of HbA and HbD and their dependence on pH, temperature and inositol polyphosphate are presented and discussed with reference to molecular structures and hypothermy that occurs during torpidity.

Amino Acid Sequence↗
PubMed1975

Hormonal regulation of lobulo-alveolar growth, functional differentiation and regression of whole mouse mammary gland in organ culture.

The entire second thoracic mammary glands of 4-week-old BALB/c female mice primed with oestradiol plus progesterone were cultivate in organ culture medium containing the "growth-promoting" hormone combinations: insulin, prolactin, growth hormone, oestradiol, progesterone and aldosterone or insulin, prolactin and aldosterone. Full lobulo-alveolar development was induced after 5-6 days of incubation and could be maintained for 15-16 days in organ culture in medium containing either hormone combination. After the initial 5-6 days in the "growth-promoting" medium, subsequent cultivation of the glands in a medium with the "lactogenic hormones", insulin, prolactin plus cortisol, led to accumulation of "milk-like" secretory material in the ductal and alveolar lumina. Incubation of the lobulo-alveolar gland in medium with insulin alone for 7-9 days resulted in complete regression of the alveoli leaving only a ductal parenchyma. Incubation in insulin, prolactin, growth hormone or insulin plus the steriod hormones for 7-9 days led to considerable alveolar degeneration without a complete regression. The results indicate that both pituitary and steroid hormones are essential for development and maintenance of mammary alveoli; insulin can only sustain the basal ductal structure.

Aldosterone↗