ALBERT

Notes: Certain monoclonal antibodies interact with proteins of Tetrahymena thermophila found in the conjugation junction as well as around the gametic nuclei (pronuclei) of conjugating cells; they also react with the oral primordium and fission zone of vegetative cells and with the cytoproct and contractile vacuole pores of all cells. One of these (FXIX-3A7) was investigated in detail. Immunogold labelling suggests that the material labelled by the 3A7 monoclonal antibody, which we call “fenestrin,” is located beneath the epiplasm (membrane skeleton). Immunoblots reveal that the major and perhaps sole antigen is a 64 kDa polypeptide, found in two isoelectric variants. Developmental studies implicate fenestrin in two processes involved in conjugation. The first is “tip transformation.” During preliminary starvation (“initiation”), labelling of fenestrin first appeared as a spot at the anterior end of starved mature cells, then after mixing of different mating types (“costimulation”) it extended posteriorly along the anterior suture. After pairing, this region spread to form a widened plate. The second process is pronuclear transfer. Fenestrations representing channels between the conjugating cells began to appear 0.5 to 1 h after the conjugants united, and eventually merged to form a small number of temporary large holes during exchange of the transfer pronuclei. A fenestrin envelope also enclosed both the transfer and resident pronuclei; a strand of fenestrin connected the two. Shortly after pronuclear transfer, both transfer and resident pronuclei were released from fenestrin caps and fused to produce a zygotic nucleus (synkaryon) not associated with fenestrin. Fenestrin thus appears to be intimately involved in the process of pronuclear exchange.

Notes: This past-presidential address considers the relationship between small-scale assembly and large-scale patterning in ciliates. Certain model examples of assembly-based patterning are reviewed, followed by the introduction of Williams and Honts' concept of “meta-assembly” as applied to the development of the oral apparatus. The major part of this review then explores two topics: the nature and origin of large-scale circumferential order, and the manner by which the large-scale order of cell directions influences the organization of membranelles of the oral apparatus. In this review, I summarize an existing formal description of large-scale positional order, allude to a more precise abstract theoretical model, and end with a brief discussion of the problem of searching for molecular mechanisms.

Notes: This paper describes the cortical anatomy and development of mirror-image doublets of Stylonychia mytilus, analyzed using the protargol technique. The reversed, or “left-handed” (LH) component of these doublets is a mirror image of the normal or “right handed” (RH) component with regard to the arrangement of cortical structures. The mirror-image patterning is imperfect, however, as the individual ciliary structures of the LH component all are of normal internal asymmetry, and the orientation of membranelles is inverted. Certain structures that would be expected to form near the line of symmetry are absent. During cell division and cortical reorganization, ciliary primordia arise and become arranged in a mirror-image pattern that is more perfect than that exhibited by the mature structures. Deviations from a mirror-image pattern appear at late stages when organelle sets differentiate within ciliary primordia: for example, the membranelle set differentiates within the oral primordium of the LH component in a sequence that is an inversion rather than a mirror image of the corresponding sequence of the RH component. This mixed control of oral development by different cortical “informational systems” accounts for some of the characteristic abnormalities of the mature oral structures of the LH component.

Notes: . Twelve monoclonal antibodies were raised that are specific for the membrane skeleton of Tetrahymena. Five were directed against T. pyrifomis and seven were directed against T. thermophila. Some cross-reactivity between species was found. Each monoclonal antibody recognized one of the three major components of epiplasm, i.e. the bands A, B, and C identified in electrophoretic separations of epiplasmic proteins. It was found, using these antibodies, that the epiplasmic proteins A, B, and C have overlapping but independent distributions within the cell.

Notes: . Vance Tartar (1911-1991) has made major discoveries concerning morphogenesis, patterning, and nucleocytoplasmic relations in the giant ciliate Stentor coeruleus, mostly by means of hand-grafting using glass microneedles. This article provides a chronological account of the major events of Vance Tartar's life, a brief description of some of his major scientific achievements, and a discussion of his distinctive personality and multifaceted interests. It concludes with a consideration of how his unique style of life and work contributed to his equally unique seientific contributions.

Notes: Specimens of Glaucoma chattoni lacking in oral areas and unable to divide were observed in both axenic and bacterized cultures. The frequency of incidence of astomy rose in the course of continued cultivation of G. chattoni A, which was the strain most extensively studied.Observations on silver impregnated animals from mass cultures, and studies of individually isolated living ciliates, have both led to the conclusion that all cells destined to become astomous pass through a certain definite sequence of events. These axe: 1. Initial damage, not microscopically detectable, predisposes the cell in such a way that within a few generattons all of its progeny become astomous. 2. The ability to complete normal stomatogenesis is lost; the cell cannot maintain normal posterior oral anlagen and also cannot regenerate following damage to the existing oral area. At this stage the cell can still divide, so that astomous opisthes are produced. 3. The existing oral area disappears shortly after the ability to produce new oral areas is lost. Loss of ability to divide occurs simultaneously with loss of oral areas. 4. Finally, there is increasing disorganization of the kineties, gradually decreasing activity and death.The incidence of astomy was compared in several different strains of G. chattoni, including two newly isolated from nature.

Notes: The techniques of silver impregnation and of digitonin solubilization have been applied to an investigation of aspects of cortical morphogenesis in Glaucoma chattoni. The typical stages of the formation of new oral areas (stomato-genesis), as well as of other related surface events, are described in detail. Estimates were made of the relative and absolute durations of these stages; these estimates indicate that for animals growing at 25°C. in bacterized culture, stomatogenesis and cell division together occupy about one third of the division cycle.Certain atypical morphogenetic phenomena have been encountered and are briefly described. These include three cases of stomatogenesis along a kinety other than kinety 1, several instances of anterior stomatogenesis leading to replacement of existing oral areas, and numerous cases of spontaneous loss of oral areas.Finally, the process of increase in number of kinetosomes along certain kineties has been submitted to intensive study. It has been found that comparatively little increase takes place during stomatogenesis and cell division; the bulk of kinetosomal increase along the kineties takes place during the first two-thirds of the division cycle. Data relevant to this increase have been analyzed according to the method of Scherbaum & Rasch; this analysis reveals that the probable phases of most rapid kinetosomal increase, along kinety l at least, occur just after division, and just before the beginning of stomatogenesis.

Notes: Mirror-image doublets of Stylonychia mytilus include 2 sets of cortical structures, one with the normal “right-handed” (RH) arrangement, the other with a reversed “left-handed” (LH) arrangement. These sets, however, are incomplete, with certain structures, most notably cirri of the right marginal type, missing near the line of symmetry. When a mirror-image doublet is bisected longitudinally to separate the RH and LH components physically, each fragment undergoes a regeneration process that restores a complete set of cortical structures, including the previously missing cirri of the right marginal type. In the resulting LH cell, all ciliary structures are present in an arrangement that is globally reversed in relation to that found in RH cells; in particular, marginal cirri of the left-marginal type are formed at the cell's right margin, and marginal cirri of the right-marginal type are produced at the cell's left margin. Whereas the regenerated RH fragment always divides and initiates a clone of normal singlets, the LH fragment, though structurally nearly complete, in all cases eventually dies without dividing. The cause of death is starvation due to the formation of an abnormal oral apparatus. In the Discussion, we consider the nature and consequences of a reversal of global positional information.

Notes: We have investigated oral development in a non-genically derived left-handed (LH) form of Tetrahymena thermophila, in which the large-scale asymmetry of arrangement of cortical structures is reversed whereas the local asymmetry of ciliary architecture remains normal. Approximately 1/2 of the oral apparatuses (OAs) of LH cells develop in the form of superficial mirror-images of OAs of RH cells. In most of these OAs, membranelles are assembled from the cells’anterior to posterior. Nonetheless, the posterior ends of these membranelles undergo the basal body displacements that lead to a “sculptured” appearance, so that the membranelles of LH OAs become organized as rotational permutations of membranelles of normal RH OAs. Many of these membranelles re-orient to a normal orientation near the end of oral development. Membranelles and undulating membranes (UMs) may develop independently of each other, and formation of postciliary microtubules of UMs is separate from that of ribbed wall microtubules. In some cases, the entire OA develops and remains as a 180° rotational permutation of the normal, resembling the inverted OAs of mirror-image doublets and LH cells of Glaucoma scintillans described by Suhama [36, 37]. We present a model (Fig. 37) for these complex developmental outcomes. These developmental patterns resemble those described previously and less completely for “secondary” OAs of cells with mirror-image global patterns, including janus cells. The present study demonstrates that such alterations in oral development are not a direct outcome of genotypic changes.

Notes: . The sequence of formation and ciliation of basal bodies and the subsequent organization of compound ciliary structures of the oral apparatus of Tetrahymena thermophila was reanalyzed with the aid of scanning electron microscopy of cells in which the epiplasmic layer was exposed, as well as by light microscopy of protargol-impregnated specimens. This combination of methods allowed the delineation of numerous steps in the patterning of the oral ciliature, some of which have received little or no previous attention. Highlights include: the initial formation of “strings” of nonciliated new basal bodies in juxtaposition to relatively few basal bodies of the stomatogenic kinety; generation of basal body pairs, roughly oriented along the anteroposterior axis of the cell, that later align side-by-side to assemble promembranelles; condensation and reorientation of promembranelles simultaneous with addition of a third row of basal bodies anterior to the original two rows; production of a very short fourth row of basal bodies at the anterior right end of each developing membranelle; generation of the outer basal body row of the undulating membrane (UM) after alignment of the inner row, with transient ciliation of the inner row preceding permanent ciliation of the outer row; limited basal body resorption at the ends of membranelles; and sculpturing of the right ends of membranelles by a movement of basal bodies associated with formation of the ribbed wall adjacent to the UM. In the old anterior oral apparatus a repetition of the processes of generation of a new outer UM row and sculpturing of right ends of membranelles takes place in synchrony with the corresponding events in the oral primordium, following prior shedding of the old outer UM row and loss of the sculptured pattern in association with temporary regression of the ribbed wall micro-tubules. Oral development is complex, with different processes involved in the assembly of the membranelles and the UM, and with a sequence of distinct events involved in the generation of each of these structures. Speaking comparatively, membranelle development follows the same pathway in many, perhaps all, ciliates in which these structures or their homologues develop from a common stomatogenic field.