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C J Flickinger

Publications and source records attributed to C J Flickinger.

At least 91 records · Page 5Linked to original sources
PubMed1982

Redistribution of material labelled with [3H]mannose in amoebae induced to undergo pinocytosis.

The synthesis, transport, and disposition of material labelled with [3H]mannose were studied by electron microscopic radioautography in normal amoebae and in cells that had internalized cell surface as a result of being induced to undergo pinocytosis. Control amoebae were injected with the precursor and placed in normal medium. The Golgi apparatus and rough endoplasmic reticulum were heavily labelled at the earliest intervals, while radioactivity of the cell surface peaked 12 h after injection of precursor. The experimental cells were injected, placed in bovine serum albumin solution from 15 to 60 min after injection, and then removed to normal medium until fixation. Incorporation of the precursor into the rough endoplasmic reticulum was near normal, but the proportions of grains associated with the Golgi apparatus and the cell surface were greatly reduced. The percentage of grains overlying vacuoles increased 12 h after injection, notably in the case of polymorphous vacuoles and dense vacuoles, both of which were identified as lysosomes with the acid phosphatase reaction. The results suggest that addition to the surface of components labelled with [3H]mannose was diminished following induction of pinocytosis. Incorporation of the precursor appeared to be shifted from cell surface material to lysosomal contents, possibly lysosomal enzymes. It is thought that this shift occurred in response to the need for the cell to digest unusually large amounts of endocytosed protein. Recycling of cell surface under these conditions is considered possible.

Acid Phosphatase↗
PubMed1981

The appearance of carbohydrate-rich material in the developing Golgi apparatus of amoebae.

The silver proteinate reaction was used to stain carbohydrate-rich substances in normal Amoeba proteus and in the developing Golgi apparatus of renucleated amoebae. Normal cells contained stained material, which probably is glycoprotein, in the cell surface, cisternae at the concave pole of the Golgi apparatus, and cytoplasmic vesicles and vacuoles. Previous radioautographic studies had shown tht glycosylation occurs in the Golgi apparatus, and that material in the Golgi apparatus is precursor to the cell surface. Amoebae were enucleated for 5 d, which results in a decline of the Golgi apparatus, the disappearance of the glycoprotein-containing cisternae preceding that of the rest of the organelle. A new nucleus was then transplanted into the enucleate amoebae, bringing about the regeneration of the Golgi apparatus. small curved cisternae that appeared 30 min after renucleation lacked staining with silver proteinate. By 1 h after renucleation, however, the content of cisternae toward the concave poles of Golgi bodies stained with silver proteinate. The Golgi apparatus in cells fixed 6 h and 1 d after operation resembled that of normal amoebae in both morphology and staining pattern. The results suggest that the developing Golgi apparatus acquired the capacity to participate in assembly of cell-surface material within 1 h after renucleation. This occurred before development of the normal enzymic activity of the Golgi apparatus was completed.

Amoeba↗
PubMed1980

Morphological characteristics of cells with apical nuclei in the initial segment of the adult rat epididymis.

The cytology of epithelial cells with apical nuclei in the initial segment of the rat epididymis was studied with the light and electron microscopes. Two types of cells were distinguished and were designated apical cells and narrow cells. The apical cells are more numerous than the narrow cells and closely resemble principal cells except for the location of the nucleus. They probably correspond to the apical cells of Reid and Cleland ('57) and may represent a variation of the principal cell. The narrow cells differ markedly from the apical cells in both light microscopic appearance and fine structure. Narrow cells strain intensely with toluidine blue and are characterized by a slender shape, many mitochodria with tubular cristae, and a large number of apical cup-shaped cytoplasmic vesicles. The possible relationship of narrow cells to other cell types is discussed.

Animals↗
PubMed1980

Capacity of enucleated amoebae to recognize and respond to heterologous cytoplasm.

When amoebae are injected with heterologous cytoplasm from another type of amoeba, cellular organelles are segregated and degenerate within membrane-bounded spaces in the host's cytoplasm. In the present study, the role of the nucleus in this response was tested by injecting heterologous cytoplasm from Pelomyxa carolinensis into enucleate and enucleate Amoeba discoides. Samples were prepared for ultrastructural study at intervals between 15 min and 2d after the operation. In enucleate recipients of heterologous cytoplasm, organelles and cellular debris were observed in vacuole-like spaces indistinguishable in morphology and frequency from those of nucleated recipients. When donor cytoplasm included thoria particles, the electron-dense tracer was found along with degenerating organelles in the vacuole-like structures, confirming the presence there of injected cytoplasm. changes of comparable magnitude were not observed when either nucleated or enucleated cells were injected with homologous cytoplasm. The results indicate that nuclear activity is not necessary for the ultrastructurally observable responses of amoebae to heterologous cytoplasm. It is suggested therefore that the cytoplasm possesses a mechanism for recognition of at least some types of heterologous organelles.

Amoeba↗
PubMed1980

Structural responses of amoebae to the injection of heterologous cytoplasm.

Responses to the introduction of heterologous cytoplasm and the fate of foreign organelles were investigated in amoebae. Heterologous cytoplasm was transferred by microinjection from Pelomyxa carolinensis to Amoeba discoides. In control experiments, homologous cytoplasm was transferred from one A. discoides to another. Recipient cells were observed by light microscopy, and samples were prepared for ultrastructural study at intervals between 15 min nad 3 days after operation. Recipients of heterologous cytoplasm showed two main responses. First, about 40% recipients expelled small amounts of cytoplasm by a blebbing process within 30 min after injection. Second, organelles were segregated and broken down in membrane-bounded cytoplasmic vacuoles between 6 h and 2 days after operation. Acid phosphatase reaction product was observed in these vacuoles along with altered organelles. Use of electron-dense thoria particles to mark donor cells demonstrated the presence of injected cytoplasm in the vacuoles. In contrast, when amoebae were injected with homologous cytoplasm, none was expelled and vacuoles containing degenerating organelles were uncommon. The survival rate and general appearance of recipients of heterologous cytoplasm were much poorer than those of homologous recipients, and most of the former died by I week after operation. It is postulated that amoeba are capable of recognizing heterologous organelles introduced into the cytoplasm and that they respond by expulsion and/or destruction of the foreign cellular components. The previously described lethal effect of heterologous cytoplasm was confirmed.

Amoeba↗
PubMed1980

Distribution of intracellular and surface components labeled with mannose-3H in amoebae exposed to the local anesthetic lignocaine.

Following injection with mannose-3H, which is a precursor to cell surface components, amoebae were exposed to the local anesthetic, lignocaine. Electron microscope radioautographs were prepared at intervals between 1 and 24 h after injection, and the distribution of silver grains over various parts of control and treated cells was determined. Control amoebae displayed successive peaks of radioactivity associated with the Golgi apparatus, cell surface, and "fringed" vacuoles. In the presence of lignocaine, incorporation of precursor into the Golgi apparatus and endoplasmic reticulum at the early intervals was inhibited, a result suggesting that glycosylation of surface components diminished. At later times, the proportion of grains associated with the cell surface and fringed vacuoles of treated cells decreased while that over lysosomes increased compared to controls. The changes in patterns of labelling of the Golgi apparatus and cell surface in lignocaine-treated amoebae resembled alterations previously observed in amoebae exposed to a general anesthetic. However, the effects of local and general anesthetic agents on the endoplasmic reticulum, lipid droplets, and lysosomes differed.

Amoeba↗
PubMed1979

Development of cell types and of regional differences in the postnatal rat epididymis.

The development of cell types and regional differences in the rat epididymis was studied in specimens of the initial, middle and terminal segments prepared at intervals between birth and postnatal day 94. The development of the epididymis was divided into three phases: (1) an undifferentiated period; (2) a period of differentiation, and (3) a phase of expansion. During the undifferentiated period, from birth to day 15, the epithelial cells had a uniform appearance. Halo cells, which are believed to be migratory leukocytes, appeared on day 14. The period of differentiation extended from day 16 to day 44. Slender, densely staining cells, termed narrow cells, appeared in the epithelium of all three segments on day 16, constituting the first evidence of differentiation of cell types in the epididymal epithelium per se. In addition to their shape and apical nuclei, the narrow cells were distinguished from other epithelial cells by the presence of cup-shaped apical vacuoles and mitochondria with tubular cristae. Principal cells and basal cells were identified on day 28, which also marked the firsh distinction of differences in epithelial height among the different segments. Narrow cells persisted into the adult in the initial segment. In the middle and terminal segments, however, narrow cells disappeared by day 35, when light cells made their appearance. The major event of the period of expansion, from day 45 to 3 months, was the appearance of sperm in the lumen between days 45 and 52. A model for differentation of cell types in the epididymis is proposed and it is suggested that narrow cells are precursors to light cells in the middle and terminal segments. The development of ultrastructural features of adult cell types preceded the appearance of sperm in the lumen.

Animals↗
PubMed1979

The effects of vasectomy on testicular metabolism in the guinea-pig.

The effects of vasectomy on testicular DNA production were studied in guinea-pigs at 2 weeks and 2 and 6 months after surgery. By 6 months, vasectomy had resulted in lower testicular weight and absolute DNA, RNA and protein contents, although RNA and DNA concentrations/protein were unchanged. Incorporation of thymidine into the testis had decreased by 2 weeks after vasectomy, and continued to do so unless the ductal system subsequently ruptured and formed a granuloma when thymidine incorporation returned to normal control levels. It is suggested that in the guinea=pig vasectomy alters spermatogenesis by reducing testicular DNA synthesis.

Animals↗
PubMed1978

The pattern of appearance of enzymic activity during the development of the Golgi apparatus in amoebae.

The appearance of enzymic activity during the development of the Golgi apparatus was studied by cytochemical staining of renucleated amoebae. In cells enucleated for 4 days, there was a great decline in size and number of Golgi bodies, or dictyosomes. Subsequent renucleation by nuclear transplantation resulted in a regeneration of Golgi bodies. Samples of amoebae were fixed and incubated for cytochemical staining at intervals of 1, 6, or 24 h after renucleation. Enzymes selected for study were guanosine diphosphatase (GDPase), esterase, and thiamine pyrophosphatase (TPPase). All three were found in the Golgi apparatus of normal amoebae but they differed in their overall intracellular distribution. GDPase was normally present at the convex pole of the Golgi apparatus, in rough endoplasmic reticulum, and in the nuclear envelope. In amoebae renucleated for 1 h, light reaction product for GDPase was present throughout the small stacks of cisternae that represented the forming Golgi apparatus. By 6 h following the operation GDPase reaction product was concentrated at the convex pole of the Golgi apparatus. Esterase, which was distributed throughout the stacks of normal Golgi cisternae, displayed a similar distribution in the forming Golgi bodies as soon as they were visible. TPPase was normally present in the Golgi apparatus but was not found in the endoplasmic reticulum. In contrast to the other enzymes, TPPase reaction product was absent from the forming Golgi apparatus 1 and 6 h after renucleation, and did not appear in the Golgi apparatus until 24 h after operation. Thus, enzymes held in common between the rough endoplasmic reticulum and the Golgi apparatus were present in the forming Golgi apparatus as soon as it was detectable, but an enzyme cytochemically localized to the Golgi apparatus only appeared later in development of the organelle. It is suggested that Golgi membranes might be derived from the endoplasmic reticulum and thus immediately contain endoplasmic reticulum enzymes, while Golgi-specific enzymes are added later in development.

Amoeba↗
PubMed1977

Relationships between membranous organelles in amoebae studied by electron microscopic cytochemical staining.

The intracellular location of a variety of enzymes was studied in Amoeba proteus with the use of electron microscopic cytochemical methods, in an attempt to assess the relationships between different membranous organelles. One group of enzymes, including nucleoside diphosphatases (IDPase, UDPase, GDPase, ADPase), carbamoyl phosphatase, alkaline phosphatase, and BAXD oxidase was localized mainly in the rough endoplasmic reticulum, nuclear envelope, and convex side of the Golgi apparatus. Esterase activity had a similar localization except that the Golgi apparatus was "stained" throughout most of its extent. A second group of enzymes was found in Golgi cisternae and vesicles, and in come vacuoles. This group included acid phosphatase, thiamine pyrophosphatase, and aryl sulfatase. Some enzymes previously detected in cytoplasmic membranes of other cells, including glucose-6-phosphatase, showed little or no activity in amoebae. The results suggest that there are chemical similarities and probable functional relationships between the rough endoplasmic reticulum, the nuclear envelope, and the convex side of the Golgi apparatus. On the other hand, the concave pole of the Golgi apparatus, aggregates of smooth tubules and vesicles, and the cell surface appear more closely related to one another than to the endoplasmic reticulum and the convex side of the Golgi apparatus. The cytochemical similarity between the Golgi apparatus and certain vacuoles such as food vacuoles may reflect the role of the Golgi apparatus in the formation of lysosomes. The locations of reaction products of the various enzymes in amoebae are compared with observations reported for other cell types.

Alkaline Phosphatase↗
PubMed1977

The influence of progestin and androgen on the fine structure of the male reproductive tract of the rat. I. General effects and observations on the testis.

The combination of a progestin and androgen has received attention as a possible male contraceptive. The progestin is thought to reduce gonadotropin release and suppress spermatogenesis, while the sex accessory organs and male characteristics are maintained by the simultaneous administration of testosterone. In the present study, the histology and ultrastructure of parts of the male reproductive tract of rats treated with medroxyprogesterone (Provera, Upjohn) (1 mg/100 g body weight/day) alone and combined with testosterone (15, 30, or 100 mug/100 g/day) were studied following treatment for up to 16 weeks. The testes and epididymides of rats administered Provera alone or Provera and testosterone weighed less than those of control rats. The weights of the accessory glands of rats treated with Provera were greatly reduced; it was possible to maintain them at approximately control levels by simultaneously administering sufficient testosterone (100 mug/100 g body weight/day). The fertility of some of the animals was tested by caging them with female rats, and none of the treated rats tested in this way was fertile. Similar microscopic alterations were present in the testes of animals administered Provera alone or Provera and different levels of testosterone. Spermatogonia, spermatocytes, and early spermatids were abundant in treated rats and did not show ultrastructural changes. However, many degenerating or necrotic spermatids of the cap phase (approximately stages 6-7) and later were present. Late spermatids of the acrosome and maturation phases were rare. Some necrotic spermatids were surrounded by Sertoli cells, and parts of spermatids lay within lysosome-lyke structures in the cytoplasm of Sertoli cells. Many large lipid droplets were also present in Sertoli cells of treated rats. Leydig cells were smaller in treated animals than in control rats. The results suggest that germ cells can develop up to cap phase spermatids but then undergo degeneration. These alterations in spermatogenesis may be responsible in large part for the antifertility effect of the progestin and androgen combination. Some rats were permitted to recover following the end of treatment. The microscopic appearance of the testis returned to normal within three to six weeks, although epididymal alterations persisted in some animals six weeks after the end of treatment. By 9 to 12 weeks after the end of treatment the reproductive organs had a normal microscopic appearance in all the rats studied.

Animals↗