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PubMed1997

Conservation of mitochondrial targeting sequence function in mitochondrial and hydrogenosomal proteins from the early-branching eukaryotes Crithidia, Trypanosoma and Trichomonas.

Kinetoplastid protozoa are the earliest-branching eukaryotes to possess a true mitochondrion. This organelle is host to a variety of intriguing and unique features, including RNA editing. We examined the characteristics of protein import into mitochondria of Trypanosoma brucei. Dihydrofolate reductase (DHFR) carrying a yeast mitochondrial targeting signal was correctly translocated into trypanosome mitochondria in vivo, as were DHFR fusion proteins bearing two unusually short (7-9 amino acids) presequences from trypanosomatids. The short trypanosomal targeting signals were functional in Saccharomyces cerevisiae as well, but their targeting efficiency was lower and processing was absent. Trichomonads branched even earlier than kinetoplastids in eukaryotic evolution and contain energy-generating organelles called hydrogenosomes. The origin of hydrogenosomes has been controversial, but most evidence suggests that they are related to mitochondria. Putative hydrogenosomal targeting signals from Trichomonas vaginalis are short (5-12 amino acids). Three such sequences were capable of targeting a passenger protein to mitochondria both in yeast and in trypanosomes, and one of the hydrogenosomal presequences was efficiently processed in both organisms. These findings suggest a resemblance between the import machineries of mitochondria and hydrogenosomes.

Amino Acid Sequence↗
PubMed1980

Ultrastructural and morphometric study of the Langerhans cell in the normal human exocervix.

The gross morphology, density, distribution and ultrastructure of the Langerhans cell of the normal human exocervix were investigated. Two standardized zinc-iodide-osmium (ZIO) procedures were applied to epithelial sheets as well as to tissue samples processed for both light and electron microscopy. Conventional electron microscopical techniques were also used. The epithelial sheet preparation allowed the visualization of the whole profile of the Langerhans cell. It was found that the number of cell processes and the degree of their branching varied greatly from one cell to another. The cells were tentatively grouped into five types and it is suggested that they represent different degrees of cell activity. The cells appeared unevenly distributed, but with a preferential location around the external os. In the three cases studied the cell density averaged 8.3 LC/mm2. The ultrastructural study revealed the classical fine structure of the Langerhans cell. The cell processes and their branches contained all the organelles found in the perikaryon, including Golgi complexes and Langerhans cell granules. The two ZIO procedures revealed that the complex inner organization of the granule does not differ from that in the epidermal Langerhans cell. It is concluded that the Langerhans cell is a constant component of the normal human exocervix.

Adult↗
PubMed2006

Cell structure of barb ridges in down feathers and juvenile wing feathers of the developing chick embryo: barb ridge modification in relation to feather evolution.

The present study deals with the cell structure and three-dimensional organization of barb and barbule cells within barb ridges of down feathers and juvenile feathers in the chick embryo. Juvenile feathers represent the second generation of feathers in the wing, and replace down feathers some weeks after hatching. Within the follicle of juvenile feathers, at 16-18 days of embryonic development, barb ridges are more numerous than in down feathers. Barb ridges of juvenile feathers contain more cells in their barbule and axial plates with respect to barb ridges of down feathers. This condition determines the formation of longer barbules inserted in the rami of juvenile feathers than barbules of down feathers. Barb ridges of juvenile feathers merge with the rachidial ridge so that pennaceous feathers are formed. Barbule cells are surrounded by cytoplasmic elongation from barb vane ridge cells located in the axial plate, which constitute most of the axial plate. The degeneration of supportive cells among barbule cells branching from barbs determine the formation of spaces between barbules. The study emphasizes that, in addition to the size of the dermal papilla, it is the length of barb ridges and the infiltration of barb ridge vane cells among barbule cells that determine the size and length of feathers. The knowledge of the cell structure of barb ridges allows understanding not only of how feathers develop but also gives insights into their evolution. Based on changes of the process of barb ridge morphogenesis some hypotheses on the evolution of plumulaceous and pennaceous feathers are presented. Feathers derived from the process of carving-out supportive cells within barb ridges and from the specific pattern of fusion of barb/barbule cells. This process initially produced variably branched down feathers and later, after barb ridge fusion, a rachis. From the modulation in the pattern of barb ridge formation various pennaceous feathers later evolved.

Animals↗
PubMed1989

[Fibroblast spreading in the presence of cytochalasin D: immunoelectron microscopy of the cytoskeleton].

Spreading of mouse embryo fibroblasts in the presence of cytochalasin D (1 microgram/ml) was studied using scanning electron microscopy, immunofluorescence, and electron microscopy of platinum replicas. Whereas circular lamellae were formed around the cell body during normal spreading, separate processes appeared at the cell periphery during spreading in cytochalasin-containing medium. The processes gradually elongated and branched. Cytoskeletons of fibroblasts spreading in the cytochalasin-containing medium were obtained by Triton X-100 extraction. They contained microtubules, intermediate filaments, actin "paracrystals" looking like short microfilament bundles, and patches of a meshwork-granular material. Immunogold coating of the cytoskeletons with anti-actin antibody showed that some meshwork-granular patches were decorated with gold particles, whereas the others were not. Non-actin patches were usually located on the distal ends of the processes, thus leaving behind the actin cytoskeletal components during the process growth. Another characteristic feature of this unidentified material is its usual association with the substratum and microtubules. These results suggest that the process protrusion during cell spreading in cytochalasin-containing medium may occur not due to actin polymerization as in the control cells, but due to involvement of some other non-actin cytoskeletal components. These components seem to be able to move along microtubules and to bind to the substratum.

Animals↗
PubMed1993

Scanning and competition between AGs are involved in 3' splice site selection in mammalian introns.

In mammalian intron splicing, the mechanism by which the 3' splice site AG is accurately and efficiently identified has remained unresolved. We have previously proposed that the 3' splice site in mammalian introns is located by a scanning mechanism for the first AG downstream of the branch point-polypyrimidine tract. We now present experiments that lend further support to this model while identifying conditions under which competition can occur between adjacent AGs. The data show that the 3' splice site is identified as the first AG downstream from the branch point by a mechanism that has all the characteristics expected of a 5'-to-3' scanning process that starts from the branch point rather than the pyrimidine tract. Failure to recognize the proximal AG may arise, however, from extreme proximity to the branch point or sequestration within a hairpin. Once an AG has been encountered, the spliceosome can still see a limited stretch of downstream RNA within which an AG more competitive than the proximal one may be selected. Proximity to the branch point is a major determinant of competition, although steric effects render an AG less competitive in close proximity (approximately 12 nucleotides). In addition, the nucleotide preceding the AG has a striking influence upon competition between closely spaced AGs. The order of competitiveness, CAG congruent to UAG > AAG > GAG, is similar to the nucleotide preference at this position in wild-type 3' splice sites. Thus, 3' splice site selection displays properties of both a scanning process and competition between AGs based on immediate sequence context. This refined scanning model, incorporating elements of competition, is the simplest interpretation that is consistent with all of the available data.

Animals↗
PubMed1999

Spectral and physical characterization of the inverted terminal repeat DNA structure from adenoassociated virus 2.

An oligodeoxynucleotide (ODN) that includes elements found in secondary structures at the 5'- and 3'- ends of adenoassociated virus 2 virion DNA was synthesized by ligation of three overlapping ODNs. The most stable secondary structure was calculated to be branched, with a 61 bp duplex stem, terminating in a three-way junction with 9 bp arms. The electrophoretic mobility of the ODN is slower than expected for normal duplex DNA of the same size, suggesting a bent or branched conformation. CD spectra indicate that the ITR structure is largely B form DNA, although there is a slight blue shift compared to the spectra of the isolated stem and loop elements. Thermal melting experiments indicate that the hairpin is significantly more stable than the isolated stem and loop elements. Singular value decomposition of UV spectra obtained as a function of temperature indicates that four species contribute to changes in the spectra upon denaturation, indicating that the melting is not a simple two-state process. Characterization of the branched ODN by differential scanning calorimetry permits estimation of the enthalpy of melting by a model-free analysis, yielding DeltaHcal= 614 kcal mol-1. This value agrees with the enthalpy computed for the most stable secondary structure.

Base Sequence↗
PubMed1992

Dynamic model of differentiation in Ewing's sarcoma cells. Comparative analysis of morphologic, immunocytochemical, and oncogene expression parameters.

We report the establishment of a model of neural differentiation in four well-characterized Ewing's sarcoma cell lines. This process was induced by serum-depleted medium (1% fetal bovine serum) and agents such as dibutyryl cyclic AMP and retinoic acid. The morphologic changes were characterized predominantly by the presence of neurite-like elongated processes showing varicosities and branching along their course with numerous internal filaments and electron-dense granules. Immunocytochemically, differentiation was accompanied by a considerable increase in reactivity for neural markers of several types: neuroblastic, neuroepithelial, neuroendocrine, Schwannian and even glial. In contrast, the tumor promoter, phorbol 12-myristate 13-acetate inhibited differentiation. Several morphologic changes were observed in phorbol 12-myristate 13-acetate-treated cells: the cells became smaller and rounder, were poorly adherent to substrate, by electron microscopy lacked cytoplasmic organelles, electron-dense granules or neural processes, and showed decreased expression of neural markers. Northern blot analysis was performed to establish whether there was any relationship between neural differentiation and degree of N-myc, c-myc and dbl oncogene expression. There was no N-myc oncogene expression in the mRNA of Ewing's sarcoma cells, even after neural induced differentiation. The degree of c-myc and dbl oncogene expression appeared heterogeneous, and varied with the culture condition. Based on these results, it may be inferred that Ewing's sarcoma cells in vitro display a variable neural phenotype, there being a variety of biologic responses to diverse culture media and various differentiation agents, but with no consistent effect on N-myc, c-myc and dbl oncogene expression.

Bucladesine↗
PubMed1977

Morphology and responses to light of the somata, axons, and terminal regions of individual photoreceptors of the giant barnacle.

1. The median eye of the giant barnacle, B. nubilus, comprises four large photoreceptor neurones which are visible under the dissecting microscope for almost their entire length. We have studied the structure of, and the responses to light recorded in, the somata, axons, and terminal regions of these neurones.2. The photoreceptor somata, each 40-70 mum in diameter, extend numerous light-sensitive dendritic processes whose membranes form rhabdomeric microvilli. Recordings from the soma show that dim light evokes a steady, noisy depolarization; brighter light elicits a transient depolarization which decays to a maintained plateau, followed by a hyperpolarization when the light is turned off.3. Light-induced voltage changes spread decrementally along the photoreceptor axons, which average 10 mm in length and 25 mum in diameter. In distal parts of the axon, near the presynaptic terminals, depolarizations and hyperpolarizations can be as large as 50% or more of their values in the soma.4. There is no demonstrable electrical coupling between photoreceptor neurones as shown by simultaneous recordings from two receptor somata or axons.5. Each photoreceptor axon enters the mid line commissure of the supraoesophageal ganglion, bifurcates, and arborizes in a restricted zone of neuropil in each hemiganglion. The large size of the terminal processes of these neurones and their characteristic cytoplasmic inclusions enable one to trace them with the electron microscope as they branch in the neuropil.6. The terminal processes subdivide and end in 1-3 mum diameter branches which are the sites of apparently chemical synapses. Vesicle-containing, presynaptic loci on these processes of the receptor cell are invariably apposed to two post-synaptic processes from cells as yet unidentified.

Animals↗
PubMed1992

Polyaxonal amacrine cells of rabbit retina: morphology and stratification of PA1 cells.

Polyaxonal amacrine cells are a new class of amacrine cell bearing one to six branching, axon-like processes, closely resembling the axons of Golgi type II cells found elsewhere in the central nervous system. Of the four types of polyaxonal amacrine cell that we have recognized in rabbit retina, three have been described previously in brief communications, and one is the subject of this paper. Type 1 polyaxonal (PA1) amacrine cells have larger cell bodies than most amacrine cells in Golgi preparations, averaging about 13 microns in diameter. These are typically positioned interstitially in the middle of the inner plexiform layer (IPL), although some are also found in the amacrine and ganglion cell layers. Axons and dendrites are broadly stratified in the middle of the IPL, in the vicinity of the a/b sublaminar border. Sparsely branching dendrites have a conventional appearance, branching at a narrow angle, and giving rise to smaller daughter branches, which taper gradually toward their termination. An unusual feature of the dendrites is the zig-zag course of some terminal branches. Clusters of small, pedunculated spines are common on proximal dendrites, and spines are virtually absent on axons. Axons emerge from proximal dendrites within 50 microns of the soma, and more rarely from the soma, in a tapering initial segment, commonly interrupted by one or two large swellings. Subsequent branching is at a wide angle, and the fine caliber is maintained in the transition from parent to daughter branches. The uniform thickness of the axonal branches is interrupted at intervals by boutons en passant. Although the extent of the dendritic tree is large, exceeding 500 microns in radial extent from the cell body, for cells a few millimeters distant from the visual streak, the axonal tree is much larger, and its radial extent is measured in millimeters. PA1 amacrine cells are believed to be polarized in their functional organization, with a primarily recipient dendritic tree and a primarily transmissive axonal tree. PA1 amacrine cells co-stratify with nab cone bipolar cells and with certain small tufted amacrine and ganglion cells at the a/b sublaminar border. The co-stratification of both axons and dendrites at the a/b sublaminar border of the IPL suggests that PA1 amacrine cells are important modulators of neural activity in the middle of the IPL, affecting both ON and OFF responses, and perhaps ON-OFF cells selectively.

Animals↗
PubMed2003

Schwann cell dynamics with respect to newly formed motor-nerve terminal branches on mature (Bufo marinus) muscle fibers.

A study has been made of the formation of synaptic terminals from long processes formed at the end of motor nerve branches of endplates in mature amphibian (Bufo marinus) muscle. Injection of fluorescent dyes into individual motor axons showed the full extent of their branches at single endplates. Synaptic vesicle clusters at these branches were identified with styryl dyes. Some terminal branches consisted of well separated varicosities, each possessing a cluster of functioning synaptic vesicles whilst others formed by the same axon consisted of closely spaced clusters of vesicles in a branch of approximately uniform diameter. All the varicosities gave rise to calcium transients on stimulation of their parent axon. Both types of branches sometimes possessed short processes (<5 microm long) or very long thin processes (>10 microm long) which ended in a bulb that possessed a functional synaptic vesicle cluster. These thin processes could move and form a varicosity along their length in less than 30 min. Injection of a fluorescent dye into terminal Schwann cells (TSCs) at an endplate showed that they also possessed very long thin processes (>10 microm long) which could move over relatively short times (<30 min). Injecting fluorescent dyes into both axons and their associated TSCs showed that on some occasions long TSC processes were accompanied by a long nerve terminal process and at other times they were not. It is suggested that the mature motor-nerve terminal is a dynamic structure in which the formation of processes by TSCs guides nerve terminal sprouting.

Animals↗
PubMed1997

Cell biology and molecular basis of denitrification.

Denitrification is a distinct means of energy conservation, making use of N oxides as terminal electron acceptors for cellular bioenergetics under anaerobic, microaerophilic, and occasionally aerobic conditions. The process is an essential branch of the global N cycle, reversing dinitrogen fixation, and is associated with chemolithotrophic, phototrophic, diazotrophic, or organotrophic metabolism but generally not with obligately anaerobic life. Discovered more than a century ago and believed to be exclusively a bacterial trait, denitrification has now been found in halophilic and hyperthermophilic archaea and in the mitochondria of fungi, raising evolutionarily intriguing vistas. Important advances in the biochemical characterization of denitrification and the underlying genetics have been achieved with Pseudomonas stutzeri, Pseudomonas aeruginosa, Paracoccus denitrificans, Ralstonia eutropha, and Rhodobacter sphaeroides. Pseudomonads represent one of the largest assemblies of the denitrifying bacteria within a single genus, favoring their use as model organisms. Around 50 genes are required within a single bacterium to encode the core structures of the denitrification apparatus. Much of the denitrification process of gram-negative bacteria has been found confined to the periplasm, whereas the topology and enzymology of the gram-positive bacteria are less well established. The activation and enzymatic transformation of N oxides is based on the redox chemistry of Fe, Cu, and Mo. Biochemical breakthroughs have included the X-ray structures of the two types of respiratory nitrite reductases and the isolation of the novel enzymes nitric oxide reductase and nitrous oxide reductase, as well as their structural characterization by indirect spectroscopic means. This revealed unexpected relationships among denitrification enzymes and respiratory oxygen reductases. Denitrification is intimately related to fundamental cellular processes that include primary and secondary transport, protein translocation, cytochrome c biogenesis, anaerobic gene regulation, metalloprotein assembly, and the biosynthesis of the cofactors molybdopterin and heme D1. An important class of regulators for the anaerobic expression of the denitrification apparatus are transcription factors of the greater FNR family. Nitrate and nitric oxide, in addition to being respiratory substrates, have been identified as signaling molecules for the induction of distinct N oxide-metabolizing enzymes.

Amino Acid Sequence↗
PubMed2000

A role for Timeless in epithelial morphogenesis during kidney development.

Central to the process of epithelial organogenesis is branching morphogenesis into tubules and ducts. In the kidney, this can be modeled by a very simple system consisting of isolated ureteric bud (UB) cells, which undergo branching morphogenesis in response to soluble factors present in the conditioned medium of a metanephric mesenchyme cell line. By employing a targeted screen to identify transcription factors involved early in the morphogenetic program leading to UB branching, we identified the mammalian ortholog of Timeless (mTim) as a potential immediate early gene (IEG) important in this process. In the embryo, mTim was found to be expressed in patterns very suggestive of a role in epithelial organogenesis with high levels of expression in the developing lung, liver, and kidney, as well as neuroepithelium. In the embryonic kidney, the expression of mTim was maximal in regions of active UB branching, and a shift from the large isoform of mTim to a smaller isoform occurred as the kidney developed. Selective down-regulation of mTim resulted in profound inhibition of embryonic kidney growth and UB morphogenesis in organ culture. A direct effect on the branching UB was supported by the observation that down-regulation of mTim in the isolated UB (cultured in the absence of mesenchyme) resulted in marked inhibition of morphogenesis, suggesting a key role for Tim in the epithelial cell morphogenetic pathway leading to the formation of branching tubules.

Animals↗
PubMed1999

Escherichia coli RuvBL268S: a mutant RuvB protein that exhibits wild-type activities in vitro but confers a UV-sensitive ruv phenotype in vivo.

The RuvABC proteins of Escherichia coli process recombination intermediates during genetic recombination and DNA repair. RuvA and RuvB promote branch migration of Holliday junctions, a process that extends heteroduplex DNA. Together with RuvC, they form a RuvABC complex capable of Holliday junction resolution. Branch migration by RuvAB is mediated by RuvB, a hexameric ring protein that acts as an ATP-driven molecular pump. To gain insight into the mechanism of branch migration, random mutations were introduced into the ruvB gene by PCR and a collection of mutant alleles were obtained. Mutation of leucine 268 to serine resulted in a severe UV-sensitive phenotype, characteristic of a ruv defect. Here, we report a biochemical analysis of the mutant protein RuvBL268S. Unexpectedly, the purified protein is fully active in vitro with regard to its ATPase, DNA binding and DNA unwinding activities. It also promotes efficient branch migration in combination with RuvA, and forms functional RuvABC-Holliday junction resolvase complexes. These results indicate that RuvB may perform some additional, and as yet undefined, function that is necessary for cell survival after UV-irradiation.

Adenosine Triphosphatases↗
PubMed1995

Possible branching of myelinated primary afferent fibres in the dorsal root of the rat.

A combined physiological and morphological examination of rat dorsal root ganglion cells revealed branching of the central process of neurones with myelinated fibres (conduction velocity > 2 m/s; n = 24). Single shock electrical stimulation of spinal dorsal roots triggered double action potentials (early and late spike) in two dorsal root ganglion cells recorded by intracellular electrodes in the in vitro spinal cord-dorsal root ganglion preparation from 12-20 day-old rats. The action potentials had different stimulus thresholds (lower for the late spike). In one dorsal root ganglion cell the differences in delay between the two spikes inhibited the soma invasion of the second spike, when both action potentials were evoked at resting membrane potential. Depolarization of the soma membrane by DC current, injected through the electrode, recovered the blocked action potential. Membrane depolarization by GABA reduced the threshold for the early spike and changed the pattern of activation. After electrophysiological characterisation, intracellular biotin/avidin staining of the neurone revealed branching of the central axon in the dorsal root. None of the other cells, which responded with single action potentials after dorsal root stimulation showed secondary branching (n = 5). This rare observation shows that differences between the conduction velocities and activation thresholds in branches of individual dorsal root ganglion cell axons may produce block of spike invasion into the soma and perhaps the spinal terminal field of large primary afferents.

Action Potentials↗
PubMed1988

Postnatal development of cat hind limb motoneurons. II: In vivo morphology of dendritic growth cones and the maturation of dendrite morphology.

The maturation of dendrite morphology was studied by light and electron microscopy in cat spinal alpha-motoneurons intracellularly labeled with horseradish peroxidase. Alpha-motoneurons supplying the triceps surae (TS) and the intrinsic foot sole (SP) muscles were investigated in kittens from birth to 44-46 days of postnatal (d.p.n.) age. At birth, a large number of dendritic branches displayed growth cones, filopodia, and fusiform processes. The growth cones were of lamellipodial and filopodial types, but intermediate forms also occurred. The growth cones shared several morphological features with the neuritic growth cones studied in vitro. It was suggested that the occurrence of different types of growth cones--even in the same dendrite--may reflect their transformation from one type to the other and the level of growth activity could be inferred from the number and form of the growth cones. About 50-70% of the terminal branches in the dendrites of newborn kittens possessed growth cones, filopodia, and/or fusiform processes. The corresponding figure for preterminal branches was 20-30%, with a clear decrease in incidence when approaching the soma. During the period under study, most of these growth-associated processes disappeared from the dendrites so that at 44-46 d.p.n. of age only about 10% of the terminal and less than 1% of the preterminal branches had growth-associated processes. Analysis of the three-dimensional distribution of dendritic branches with such processes disclosed that they were relatively more frequent in the medial, rostral, and caudal dendritic territories. It was concluded that the pattern of distribution and disappearance of growth cones, filopodia, and fusiform processes coincided with postnatal longitudinal dendritic growth and the development of the adult dendritic territories described in a preceding paper (Ulfhake et al., '88). Dendritic growth, with respect to length and caliber, also occurred in the absence of growth cones and filopodia. It is suggested that the important role of these processes may be to act as a steering device in establishing the adult distribution and synaptology of the dendrites. Comparison of TS and SP alpha-motoneuron dendrite morphology at birth and at 22-24 d.p.n. age showed that the SP neurons lagged in the maturation process. Light and electron microscopic observations indicated that postnatally direct contacts might exist between dendrites and fine blood vessels in the neuropil without any interposing glial sheath. The number of such suspected contacts diminished during the period under study, indicating that the glial ensheathment of the blood vessel takes place, in part, postnatally.

Animals↗
PubMed1987

[Endometrial cups in horses].

Endometrial cups in horses are outgrowths appearing in the uterine wall of the pregnant horn between approximately 38 days and 150 of gestation. The cups are structures which vary in shape from oval to irregular and have distinct raised edges, showing an ulcer-like form. The maximum measurements are approximately 5 X 2.5 X 2.5 cm. Microscopic examination shows that they consist of large epithelioid decidual-like cells having large nucleoli. Pregnant Mare Endometrium Gonadotrophin (PMEG) is produced in the endometrial cups. PMEG may be regarded as a precursor of Pregnant Mare Serum Gonadotrophin (PMSG), which can be detected in the blood of mares. The exact function of PMSG is still obscure; some possible functions are suggested in the present paper. After approximately 70 days of gestation, regression starts. The regression of the cups is an immunological process in which the cellular branch of the immunological system plays an important role. Humoral immunity plays a more protective role in the process of regression, probably as a result of the production of 'blocking' antibodies.

Animals↗
PubMed1978

Steps involved in the processing of common precursor forms of adrenocorticotropin and endorphin in cultures of mouse pituitary cells.

The initial steps in the processing of the common precursor to adrenocorticotropin (ACTH) and endorphin in mouse pituitary tumor cells (AtT-20) have been investigated. Three forms of the precursor have been resolved by sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel electrophoresis with apparent molecular weights of 29 000 (29K ACTH-endorphin), 32 000 (32K ACTH-endorphin) and 34 000 (34K ACTH-endorphin). These forms have a similar peptide backbone, but their carbohydrate content differs. In particular, a tryptic glycopeptide has been observed in 32K ACTH-endorphin which is not present in 29K ACTH-endorphin and has been identified as the tryptic peptide containing the alpha(22--39) sequence of ACTH. Similar heterogeneity in carbohydrate has been observed in some of the smaller molecular weight forms of ACTH which are resolved by NaDodSO4 gel electrophoresis. Pulse chase and continuous labeling studies using radioactive amino acids and sugars suggest that the 29K ACTH-endorphin is converted to 32K and 34K ACTH-endorphin by the addition of carbohydrate. The glycopeptide and pulse chase studies suggest that 29K ACTH-endorphin is at a branch point in the processing pathways. It can either be converted to 4.5K ACTH by proteolytic processing or to 32K ACTH-endorphin by the further addition of carbohydrate. The 32K ACTH-endorphin can then be converted to 13K ACTH, the glycosylated form of 4.5K ACTH (Eipper, B.A., & Mains,, R.E. (1977) J.Biol. Chem.252, 882), by proteolytic processing. A comparison of the distribution of the different molecular weight forms of ACTH and endorphin in mouse pituitary extracts and in the mouse pituitary tumor cells reveals that the pituitary contains all of the forms of ACTH and endorphin seen in the tumor cells, including the three forms of the ACTH-endorphin precursor. However, the molecular weight distribution of the forms in the anterior lobe is very different from that in the intermediate lobe of mouse pituitary.

Adrenocorticotropic Hormone↗