Kiyoshi Terakado

Prolactin-like immunoreactivity in the granules of neural complex cells in the ascidian Halocynthia roretzi

Abstract.Electron-microscopic studies of the neural complex (neural gland, dorsal strand, and cerebral ganglion) of an ascidian, Halocynthia roretzi, were performed, paying particular attention to the secretory systems. We found that cells scattered along the dorsal strand and neural cells in the cerebral ganglion contained electron-dense secretory granules of variable size. Immunoelectron-microscopic studies with an antiserum to bullfrog prolactin revealed that the secretory granules (100–250 nm in diameter) of some granulated cells contained a prolactin-like substance. Cells belonging to the neural gland and dorsal strand neither contained electron-dense granules nor showed immunoreactivity. The possibility that cells in the cerebral ganglion and those along the dorsal strand are phylogenetic progenitors of vertebrate adenohypophyseal cells is discussed.

Structure of multinucleated smooth muscle cells of the ascidian Halocynthia roretzi

SummaryCells isolated from ascidian smooth muscle were about 1.5–2 mm in length. Each contained 20–40 nucle in proportion to cell length. The cytoplasm was characterized by the presence of an enormous quantity of glycogen particles, tubular elements of sarcoplasmic reticulum coupled to the cell membrane, and conspicuous contractile elements. Thick and thin filaments had diameters of about 14–16 nm and 6–7 nm, respectively. The population density of the thick filaments was much higher (mean 270/μm2 filament area) than in vertebrate smooth muscles. The ratio of thick to thin filaments was about 1∶6. All the thick filaments were surrounded by a single row of 5–9 thin filaments forming a rosette, and cross-bridges with periodicities of 14.5 and 29 nm were found between them. The contractile apparatus consisted of numerous myofibrils which were arranged nearly along the cell axis and were separated from each other by a network of 10-nm filaments. The myofibrils further consisted of many irregularly arranged sarcomerelike structures, each of which was comprised of a small group of thick and thin filaments with attached dense bodies.

TEST CELL MIGRATION AND TUNIC FORMATION DURING POST-HATCHING DEVELOPMENT OF THE LARVA OF THE ASCIDIAN, CIONA INTESTINALIS

Morphological changes in the tunic layers and migration of the test cells during swimming period in the larva of the ascidian, Ciona intestinalis, were observed by light and electron microscopy. The swimming period was divided into three stages. In stage 1, further formation of juvenile tunic layer started only in the larval trunk and neck region. In stage 2, the layer became swollen in the ventral and dorsal sides of the neck region and in stage 3, the swelling expanded backward. Concomitantly with these changes, the outermost larval tunic layer (outer cuticular layer), which had been formed before hatching, also swelled in the neck region in stage 2 and formed two humps in stage 3, although the layer did not change in the tail region during the swimming period. Test cells that were present over the entire larval tunic layer in stage 1 began to move from the surface of the fin toward that of the side of the body in stage 2, and finally gathered to form six bands running radially from the anterior end to the posterior end of the trunk region and aligned along the lateral sides of body in the tail region in stage 3. In electron microscopic observations, pseudopodia protruding from the test cells invaded the larval tunic, following which they extended proximate to the juvenile tunic in the trunk region. In the tail region, which had no juvenile tunic layer as that described, the pseudopodia invaded and remained adjacent to the surface of the epidermis or the sensory cilia protruded from the epidermis. Metamorphosis of the larvae, further tunic formation, degradation of adhesive papilla, attachment of larva to the substratum and tail resorption commenced after these morphological changes occurred. The possible role of the test cells in metamorphosis is discussed.

CYTOLOGICAL AND ULTRASTRUCTURAL STUDIES ON MUSCLE DIFFERENTIATION IN THE ASCIDIAN, PEROPHORA ORIENTALIS*,**

Muscle cell differentiation in the tail of the ascidian, Perophora orientalis, from early tail‐bud embryos to swimming larvae, were studied cytologically and ultrastructurally. Myogenic cells did not form multinucleated myotubes, but remained as mononucleated cells. Nucleolar component increased prior to a marked increase in cytoplasmic RNA. Cytoplasmic RNA appeared first around nucleus and later concentrated in the peripheral cytoplasm. The fine filaments measuring 20–30 Å in their thin parts and 30–45 Å in their thick parts in diameter appeared initially, forming loose networks, in the peripheral cytoplasm where ribosome clusters had been concentrated. These filaments were tightly attached by particles of various size and density. These filaments tended to be arranged in parallel as they increased in their size. They seemed to be precursors of both actin and myosin filaments of formed myofibrils. Z band precursors were found as dense patches in association with loosely arranged myofilaments and consisted of particulate and filamentous materials. The myofibrils seemed to grow further by organizing free filaments into bundles and further by aligning bundles of myofilaments at both ends.

Ultrastructure of the thread cells in the slime gland of japanese hagfishes paramyxine atami and eptatretus burgeri

SummaryThe thread cells in the slime gland of Japanese hagfishes, Paramyxine atami and Eptatretus burgeri were studied by light and electron microscopy.The mature thread cells are large elements (180×80 μ) filled with an intricately coiled thread, approximately 2 μ in diameter. The protein nature of the thread has been confirmed by histochemical examination. In the initial stage of growth, the thread consists of a bundle of distinctly parallel filaments approximately 90–120 Å in diameter and a centrally located tubular component approximately 230–260 Å in diameter which occurs singly or occasionally as a double and triple structure. The developing thread displays thin filaments, approximately 30–60 Å in diameter. The thin filaments are composed of fine fibrous structures, subfilaments, approximately 10–30 Å in diameter. On the outer surface of the thread a coating is apparent, giving it a fluffy appearance. Polysomal clusters consisting of five or six ribosomes are predominant. Fine fibrous structures are also found among the threads. They seem to have a spatial relationship with the polysomes and resemble the subfilament constituents of the thin filaments.From these results, it may be suggested that the fine fibrous structures synthesized by polysomes, twist together and coalesce into a thread. The problem of the polysome size and the molecular weight of the fibrous protein synthesized is discussed.

Impaired insulin secretion from the pancreatic islets of hypothyroidal growth-retarded mice

The growth-retarded (grt) mouse shows thyroid dysfunction-related hyporesponsiveness to TSH. Thyroid hormone is a critical regulator of metabolism in many cells; thus, derangement of thyroid function affects many organs and systems. Experiments were conducted focusing on the function of the pancreatic islets in grt mice. We showed occurrence of a fasting hyperglycemia and a decreased plasma insulin level response to a glucose load in grt mice, despite normal insulin molecules being stored in secretory granules of pancreatic islets. We also demonstrated a reduction of insulin secretion in response to glucose administration from islets of grt mice in vitro, while the insulin release in response to KCl stimulation was comparable to that in normal mice, indicating that the isolated islets from grt mice have normal ATP-sensitive K(+) channels and postchannel activity. The mRNA expression levels of glucose transporter 2 and glucokinase in the islets of grt mice were similar to those in normal mice. Triiodothyronine administration to grt mice improved insulin secretion very slightly. On the other hand, mRNA for tyrosylprotein sulfotransferase 2 (Tpst2) was found to be expressed in the pancreatic islets of grt mice. Considering that Tpst2 is the responsible gene of grt mice, mutation of which is associated with a poor function of TSH receptor, the findings raise a possibility of involvement of factors including Tpst2 in the insulin hyposecretion in grt mice.

THE EFFECTS OF ACTINOMYCIN D ON MUSCLE CELLS OF ASCIDIAN EMBRYO

The effect of actinomycin D on muscle cells development of the ascidian, Herdmania momus was studied ultrastructurally. No myofilament was formed when the drug was given at any stage before early tail‐bud stage (stage 3). Some aggregates of myofilaments in various size were formed when the treatment was started at stage 4 (4.5 hr after fertilization at about 28°C). Above 60% of myofibrils of fully differentiated muscle cells were formed when the treatment was initiated at stage 5 (5 hr after fertilization). Muscle cells of the tadpoles treated from stage 7 (6 hr after fertilization), at which myofilaments were first detectable in normal development, differentiated almost normally. It is therefore suggested that most mRNAs for muscle proteins are synthesized preceding the onset of myofilament formation and are relatively stable. It is also suggested that mRNAs for myosin, actin and Z band materials are almost simultaneously synthesized.

Occurrence of prohormone convertase-like substances in the neural complex cells of the ascidian Halocynthia roretzi.

Our previous study on the distribution of adrenocorticotropin (ACTH)-like substances in the neural complex (cerebral ganglion, dorsal strand, and neural gland) of an ascidian Halocynthia roretzi revealed that some of the cells in the cerebral ganglion and the cells scattered along the dorsal strand were immunopositive with antiserum against ACTH. In order to ascertain whether these cells are equipped with prohormone convertases, we performed immunohistochemical studies on the neural complex by using antisera against PC1 and PC2. A considerable number of cells around the dorsal strand and a few cells in the neural ganglion were immunopositive with PC1 and/or PC2 antibodies. Immunoelectron microscopic study demonstrated that some granulated cells situated in the cerebral ganglion and along the dorsal strand contained PC1- or PC2-like substances within their secretory granules. Western blot analysis revealed the presence of 66-kDa PC1-like and 70-kDa PC2-like substances in the neural complex. Moreover, immunostaining of consecutive sections showed that the majority of the cells containing PC1- and/or PC2-like substances corresponded to the cells immunoreactive with antisera against ACTH and CLIP but not to those immunoreactive with an antiserum against PRL. Cells belonging to the neural gland neither contained electron-dense granules nor showed immunoreactivity with any antisera employed in this experiment. The possibility that some of the cells situated in the cerebral ganglion and along the dorsal strand are progenitors of vertebrate adenohypophyseal cells is discussed.

Adrenocorticotropin-Like Immunoreactivity in the Granules of Neural Complex Cells of the Ascidian Halocynthia roretzi

Abstract Immunohistochemical studies on the neural complex (neural gland, dorsal strand, and cerebral ganglion) of an ascidian, Halocynthia roretzi, were performed by using an antiserum against porcine ACTH. The antiserum recognized a considerable number of the cells scattered along the tubular structure of the dorsal strand and a few cells in the cerebral ganglion. Immunoelectron microscopic studies revealed that the ACTH-like substance resided within secretory granules with diameter of 300–500 nm. Furthermore, those ACTH-immunoreactive cells were demonstrated to be different from PRL-immunoreactive cells, the presence of which had previously been reported.

Chromatin Arrangement and Axis Formation in the Spermiogenesis of a Pulmonate Snail

Nuclear change in relation to axis formation and condensation during spermiogenesis was investigated in the snail, Physa acuta. In the early spermatid, characteristic thick layers (termed apical and basal plates) are formed on two sides of a nuclear envelope. Soon after the formation of these plates, a developing acrosome and a flagellum attach externally to the center of the apical and basal plates, respectively. However, most (presumably all) of the chromatin filaments become attached all over the inner surface of the apical and basal plates. This means that the plates themselves are actually the specialized forms of the nuclear envelope to which chromatin filaments become connected; by means of these plates, the chromatin filaments become arranged in parallel to the antero‐posterior axis as the nucleus elongates. This suggests that the formation of these two thick layers on opposing surfaces of the nucleus primarily determines the antero‐posterior axis of the spermatid and the direction of the arrangement of chromatin.